TWI316961B - Peptides and related compounds having thrombopoietic activity - Google Patents

Description

1316961 (Ou, invention description (invention description should be stated: the technical field, prior art, content, embodiment and schematic description of the invention) Technical Field This "fx is generally related to the success of thrombocytopenic activity Skin and related compounds. The compounds of the invention may be used to increase the production of platelets or platelet precursors (such as megakaryocytes) in mammals. Prior Art The present invention relates to having in vitro and in vivo (in vivo) Compounds that stimulate the production of platelets and their precursor cells, such as megakaryocytes, particularly peptides. Two proteins known to have platelet-forming activity are proposed: thrombopoietin (TPO) and megakaryocytes Growth Development Factor (MGDF), as background. Colonization of endogenous thrombopoietin (TPO) (Lok et al., Nature 369: 568-571 (1994); Bartley et al., Cell 77: 1117- 1124 (1994); Kuter et al., Proc. Natl. Acad. Sci. USA 91:11104-11108 (1994); de Sauvage et al., Nature 369:533-538 (1994); Kato et al., Journal Of Biochemi Stry 119:229-23 6 (1995); Chang et al., Journal of Biological Chemistry 270:511-514 (1995)) has rapidly increased my megakaryocyte production (megakaryocyte production) and platelet production (platelet production) Understanding. Endogenous human TPO, a 60 to 70 kDa glycosylated protein produced in the liver and kidney, is composed of 3 3 2 amino acids (Bartley et al., Cell 77: 1117-1124 ( 1994); Chang et al., Journal of Biological Chemistry 270:511-514 (1995). The protein is highly conserved among different species and is at the amino terminus (amino acid 1 to 172) (Bartley et al) , Cell 77: 1117-016622 1316961 (2) 1124 (1994)) has a homology of 23% with human erythropoietin (Gurney et al., Blood 85: 981-988 (1995)). Derived TPO displays all of the features of a key biological modulator with platelet production. Its in vitro activity includes purified murine hematopoietic stem cells (Zeigler et al., Blood 84:4045-4052 (1994)) and human CD34+ cells (Lok et al., Nature 369:568-571 (1994). Rasko et al., Stem Cells 15:33-42 (1997)) The specificity of both megakaryocyte communities, with increased ploidy production of megakaryocytes (Broudy et al., Blood 85:402-413 (1995)) and induction of terminal megakaryocyte maturation and platelet production (Zeigler et al., Blood 84:4045-4052 (1994); Choi et al., Blood 85:402-413 (1995) ). In contrast, the synthetic antisense oligodeoxynucleotide against the TPO receptor (c-mpl) significantly inhibited the ability of megakaryocyte protoplast formation (Methia et al., Blood 82:1395) -1401 (1993)). In addition, c-mpl knockout mice have severe thrombocytopenia and lack megakaryocytes (Alexander et al., Blood 87: 2162-2170 (1996)). Recombinant human MGDF (rHuMGDF, Amgen Inc., Thousand Oaks, CA) is another TPO-associated platelet-producing polypeptide. It is produced by transforming Escherichia coli (E. coli) containing cDNA encoding a truncated protein covering the amino-terminal receptor-binding functional site of human TPO (Ulich et al., Blood 86:971) -976 (1995)). After the multi-peptide is extracted, refolded and purified, the poly[ethylene glycol] (peg) moiety is covalently bonded to the amine end. The resulting molecule is referred to herein as PEG-rHuMGDF or simply MGDF. 016623 1316961 (3) Various studies using animal models (Ulich, TR et al., Blood 86: 971-976 (1995); Hokom, Μ. M. et al., Blood 86: 4486-4492 (1995)) The efficacy of TPO and MGDF in bone marrow transplantation and in the treatment of thrombocytopenia, a condition usually caused by chemotherapy or radiotherapy, was confirmed. Preliminary data from the human body identified the use of MGDF to increase platelet count in various coagulum. (63886161 & 1., 1^11. 61 348-1279-81 (1996); Kato et al., Journal of Biochemistry 119: 229-236 (1995); Ulich et al., Blood 86: 971-976 (1995) )). MGDF can be used to enhance the platelet administration procedure because the administration of MGDF enhances the blood platelet count in the healthy platelet donor cycle system by approximately three times the original value. TPO and MGDF exert their effects through the binding of c-mpl receptors, which are mainly expressed on the surface of certain hematopoietic cells, such as megakaryocytes, platelets, CD34+ cells and primitive progenitor cells (Debili). , N. et al., Blood 85:391-401 (1995); de Sauvage, FJ et al., Nature 369:533-5 3 8 (1994); Bartley, TD, et al., Cell 77:1117- 1124 (1994); Lok, S. et al., Nature 3 69:565-8 (1994)). Like most receptors for interleukin and protein hormones, the c-mpl receptor belongs to the class I interleukin receptor superfamily (Vigon, I. et al., Proc. Natl. Acad. Sci. USA 89:5640-5644 (1992)). Activation of such receptors involves ligand binding-induced receptor homodimerization, which in turn triggers a cascade of transduction events. In general, the interaction of protein ligands and their receptors usually occurs at a larger interface. However, as evidenced by the fact that human growth hormone is linked to its receptors, only some of the key residues on the interface actually contribute most of the binding energy (Clackson, T. et al., Science 016624) Continuation page ' x J- Λ , < 1316961 (4) 267: 383-386 (1995)). This fact, which is used only with the body of the remaining protein ligands to show the correct epitopes (epitopes), can be used to find active ligands with far smaller sizes. Accordingly, only molecules with a "peptide" length (eg, 2 to 80 amino acids) can bind to a receptor protein of a given large protein ligand. Such a peptide can mimic a large protein ligand. Activity, or through competitive binding, inhibits the biological activity of large protein ligands, and is often referred to as a "peptide mimetic" or "simulated peptide." Phage display peptide libraries have emerged as recognition of such peptide mimetics A powerful technique in terms of. See, for example, Scott, J_K. et al., Science 249:386 (1990); Devlin, JJ. et al., Science 249: 404 (1990); US Patent No. 29, 1993 U.S. Patent No. 5,733,731 issued March 31, 1998; U.S. Patent No. 5,498,530 issued on March 12, 1996; U.S. Patent No. 5,432,018 issued on Jul. 11, 1995; U.S. Patent No. 5,338,665 issued to Japan, U.S. Patent No. 5,922,545, issued on Jul. 13, 1999; WO 96/40987, issued on December 19, 1996; and WO 98/1 5,833, issued on Apr. 16, 1998. (All the above documents are incorporated by reference in their entirety. In such a library, the random peptide sequence is displayed by fusion with the coat protein of the filamentous phage. Typically, the peptide shown is directed against the antibody to immobilize the extracellular functional site of the receptor. The retained phage can be enriched by a continuous cycle of affinity purification and repopulation. The best binding peptide can be sequenced to identify key residues within one or more structurally related peptide families. See, for example, Cwirla, et al., (1997), Science 276; 1696-9. The peptide sequence can also be inferred which residues can be scanned via alanine or DNA levels 016625 1316961 (5) Safe substitution. Sequences can be generated and screened to further optimize the sequence of the optimal binder. Lowman (1997), Ann Rev. Biophys. Biomol. Struct 26:401-24. Structure for protein-protein interactions The analysis can also be used to extrapolate peptides that mimic the binding activity of large protein ligands. In such assays, the crystal structure can infer the bulk and relative orientation of key residues of large protein ligands. Take a peptide. See, for example, Takasaki, et ai, (1997), Nature Biotech, 15: 1266-70. These assays can also be used to study the interaction between receptor proteins and sputum display-selected peptides. Role, which can be speculated to further enhance the peptide to increase binding affinity. There are other methods in the peptide study that compete with the phage display method. Shengyue Taiku can be fused to the carboxy terminus of the lac repressor (repress〇r) and expressed in Escherichia coli. Other E. coli-based methods can be displayed on the outer membrane of cells by fusion with peptide pegylated glycoprotein (PAL). In the following, these and related methods are collectively referred to as "E. coli display method". In other methods, translation of random RNA is halted prior to ribosome release, resulting in a multi-peptide library to which its associated RNA is still attached. Hereinafter, this and related methods are collectively referred to as "ribosome display method". Other methods employ peptides linked to RNA, such as PRO fusion technology, phyl〇s, Inc. See, for example, R〇berts &

Szostak (1997), Proc. Natl. Acad. Sci. USA, 94: 12297-303. In the following, 'this and related methods are collectively referred to as "RNA_peptide screening method." A library of chemically derived peptides has been developed in which peptides are immobilized on stable, non-biological materials, such as poly-glyphs. Or a solvent permeable resin. Another chemically derived peptide library is photolithographically scanned to be immobilized on a glass slide. Win 016626 • 10-1316961 (6) Description of the Invention Continued Peptide. In the following, this and related methods are collectively referred to as "Chemistry-Peptide Screening". The advantage of chemical-peptide screening is that it can use D-amino acids and other non-natural analogs, as well as non-peptide ingredients. Both biological and chemical methods are available at Wells & Lowman. (1992), Curr. Opin. Biotechnol, 3: 355-62. Conceptually, peptide analogs of any protein can be found using phage display, RNA-peptide screening, and other methods described above. Phage display peptide library technology and found small peptide molecules that act as agonists of the c-mpl receptor (Cwirla, SE et al., Science 276: 1696-1699 (1997)). In this type of study, random small wins Peptide sequences are shown as The fusion of the filamentous phage outer layer protein is eluted by affinity of the antibody to the c-mpl receptor for immobilization of the extracellular functional site, and the retention of the retained phage has been used for the second round of affinity purification. Combining the selection and re-proliferation methods to enrich the tighter pool of binders. As a result, the two families of c-mpl-binding peptides were first identified, which are independent of each other in their sequence. To further optimize the optimal binder, which ultimately leads to the separation of the very active peptide with IC5 〇 = 2 nM and its EC5 〇 = 400 nM (Cwirla, SE· et al., Science 276: 1696-1699 (1997) 14-residue ΤΡΟ The analog peptide does not have significant sequence homogeneity with TPO or MGDF. The structure of this specific TPO mimetic peptide (TMP) compound is as follows: lie Glu Gly Pro Thr Leu Arg Gin Trp Leu Ala Ala Arg Ala (SEQ ID NO: 1) or a single letter amino acid abbreviation denoted as IEGPTLRQWLAARA. In a previous similar study of EPO mimetic peptides, an EPO mimetic peptide (EMP) was discovered using the same technique (Wrighton, NC et al_) , Science, -11 - 〇 16627 1316961 (7) Description of the Invention (Continued from 273: 458-463 (1996)), and found that the peptide acts in the form of a dimer when bound to the EPO receptor (EPOR). Based on X-ray crystallographic data, the (ligand) 2/(acceptor) 2 complex thus formed has C2 symmetry (Livnah, 0. et al., Science 273:464-471 (1996)). Based on this structural information, a covalently linked EMP dimer was designed in which the C-termini of two EMP monomers were crosslinked with elastic spacers and found to have greatly enhanced binding and in vitro/in vivo biology. Activity (Wrighton, NC, et al., Nature Biotechnology 1 5: 1261 - 1265 (1997)). A similar C-terminal. Dimerization method was applied to the TPO analog peptide (TMP). (Cwirla, S.E. et al., Science 276:1696-1699 (1997)). The C-terminal ligation dimer (cc linkage) of a particular ΤΡ0 mimetic peptide was found to have improved 0.5 nM binding affinity in cell proliferation assays, and increased in vitro activity (EC50 = 0.1 nM) (Cwirla, SE et al., Science 276: 1696-1699 (1997)). The utilization of recombinant proteins for therapeutic use leads to advances in protein modification to enhance or improve the properties of such proteins as pharmaceutical agents. Such modifications provide enhanced protein protection and reduced degradation by reducing or eliminating protein hydrolysis. Other advantages include increased stability, cycle time, and biological activity of therapeutic proteins under certain circumstances. A review article explaining protein modification is Francis, Focus on Growth Factors 3:4 -10 (May 1992) (published by Mediscript, London, UK). Useful modifications of protein therapeutics include linkage to polymers such as polyethylene glycol (PEG) and dextran. Such modifications are discussed in detail in the following patent applications: "Modified Peptides as Therapeutic Agents", US Patent -12-016628 1316961 (8) Issued with, Cheng Ming tribute: 'Application No. 09/428,082, PCT Application w〇 00/24782, the entire disclosure of which is incorporated herein by reference. Another such modification is the use of the Fc region of the immunoglobulin molecule. The antibody comprises two functionally independent fractions; a variable functional site, called " Fab", which binds antigen, a fixed functional site called "FC", which provides linkage to effector functions such as complement or phagocytic cells. The Fc fraction of immunoglobulin has a long-lived half-life, while the Fab is short-lived. (Capon, et al Nature 337, 525-531 (1989)). Fc functional sites have been used to constitute therapeutic protein products to provide longer half-life or to add function 'such as Fc receptor binding, protein A binding, complement fixation and placental transplantation, all of which are present in the Fc protein of immunoglobulins. . (Capon, et al., Nature 337: 525-531 (1989)). For example, the Fc region of an IgG1 antibody is fused to CD30-L, which binds to CD30, which is expressed in Hodgkin disease tumor cells, undifferentiated lymphoma cells, T-cell leukemia cells, and other malignant cell types. body. See U.S. Patent No. 5,480,981. IL-10, an anti-inflammatory and anti-rejection agent, is fused to the fusion mouse Fc 2a to increase the short-cycle half-life of interleukins (Zheng, X et al., The Journal of Immunology, 154: 5590-5600 (1995) ). A number of studies have also evaluated the use of tumor necrosis factor receptors to attach Fe proteins to human igG1 for the treatment of patients with septic shock (Fisher, C. et al N Engl. J. Med., 334:1697-1702 ( 1996); Van Zee, K. et al The Journal of Immunology, 156:2221-2230 (1996)). There are also therapeutic molecules that use the & CD4 receptor to produce AIDS. See CapM et al., Nature, 337: 525-53 1 (1989). In addition, there is also the incorporation of interleukin 2 into the 13-invention 'Note: Spray page 1316961 (9) to the Fc portion of IgG1 or IgG3 to overcome the shorter half-life of interleukin 2 and its systemic toxicity. See Harvill et al., Immuno technology, 1:95-105 (1995). A specific platelet-causing compound, generally a peptide, having a cascade (ie N- to C-terminus) orientation and a cascade of peptide dimers linked at its N-terminus is disclosed in published PCT application WO 00/24770. For carrier molecules, such as linear polymers, oligosaccharides or Fc groups. There is a need to propose additional compounds which have excellent biological activity to stimulate the production of platelets (thrombotic activity) and/or the production of platelet precursor cells, particularly megakaryocytes (megakaryocyte production activity). There is also a need to propose a compound which exhibits platelet-forming activity and which has excellent therapeutic properties such as long-term life. Such compounds exhibit advantageous properties related to production, isolation, purification, biological activity, stability, and cycle time. The present invention proposes novel compounds and related aspects having such activities. SUMMARY OF THE INVENTION The present invention is directed to a therapeutic compound that binds to a c-mpl receptor (hereinafter referred to as "mpl acceptor"). More particularly, the present invention proposes a group of compounds which exhibit enhanced ability to bind to the c-mpl receptor and/or to transmit, ie, activate, the c-mpl receptor to trigger a transmembrane signal, the receptor and the vector The receptor for the activity of endogenous thrombopoietin (TPO) is the same. Therefore, the compounds of the present invention have excellent platelet-forming activity, i.e., can stimulate platelet production and/or megakaryocyte production activity in vivo and in vitro, i.e., stimulate platelet precursor production in vivo and in vitro. In addition, certain of the compounds of the invention also exhibit superior therapeutic properties, such as improved semi-synthesis in plasma -14-016630 1316961 (10), biological activity, and in vivo circulating time. In one aspect, the invention features a compound that binds to an mpl receptor, the compound comprising the following sequences:

X1-X2-X3-X4-GPTL-X9-X10-WL-X13-X14-X15-X16-X17-X18 wherein X1-X4, X9-X10 and X13-X18 are each independently an amine group as defined herein An acid, and wherein the compound has a binding affinity and/or biological activity to the mpl receptor greater than that of the following sequence: IEGPTLRQWLAARA. In another aspect, the invention provides a compound that binds to a c-mpl receptor having the following sequence: X1-X2-REGPTLRQWL-X13-WRR-X17-X18 wherein XI, X2, X13, X17 and X18 are each Independently an amino acid. In another aspect, the invention provides a compound that binds to an mpl receptor comprising a sequence selected from the group consisting of SEQ ID NO 2 to SEQ ID NO 30.

In another aspect, the invention is a dimer or multimer of a compound comprising a sequence selected from the group consisting of SEQ ID NO 2 to SEQ ID NO 30. In another aspect, the present invention provides a composition of matter that binds to an mpl receptor having the following general formula: (LNl)r-(TMPl)a-(LN2)m-(TMP2)b-(LN3)n- (TMP3)c-(LN4)0-(TMP4)d wherein TMP1, TMP2, TMP3 and TMP4 are each independently selected from the group consisting of TMPs disclosed herein; LN1, LN2, LN3 and LN4 are each independently As a linker; a, b, c and d are each independently an integer from 0 to 10; -15- 1316961

〇i) 1, m, n and 0 are each independently an integer of 〇2〇. In still another aspect, the invention provides a substance composition that binds to an mpl receptor, which has the general formula: (Vl)v--(LNl)1--(TMPl)a..(LN2)m--( TMP2)b--(LN3)n--(TMP3)c--(LN4)〇--(TMP4)d--(V2)w where νι and V2 are each independently a vehicle, and w is independently an integer of 〇1. 4 The compounds of the present invention can be prepared by standard synthetic methods, recombinant DNA techniques or other methods for preparing peptides and fusion proteins. The compounds of the invention encompassing the non-peptide portion can be synthesized from standard organic chemical reactions in addition to standard peptide chemical reactions. The compounds of the present invention can be used for therapeutic or prophylactic purposes by administering them together with a suitable pharmaceutical carrier material and administering to a patient, e.g., a human (or other mammal) which is in need thereof, in an effective amount. The agent _ linkages have the same activity as the peptides that are mimicked by the peptides, and the natural ligands of the day, here compared to the blood plateletin, or even larger. In another aspect, the invention provides for the treatment of A, A, and soil. The law of microplatelet reduction. In other aspects, the invention features methods of increasing , , , megakaryocytes or platelets' and methods of producing the compounds described herein. In another aspect, the invention also provides related pharmaceutical compositions. In another aspect, the invention provides a polynucleic acid which encodes a composition of matter of the present invention, including the composition of the polynucleotide, including the expression vector of the polynucleotide, and includes performance of the child. The host cell of the vector. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 shows a peptide of the present invention. FIG. 1 shows a peptide of the present invention, which is apparent from the following detailed description. An exemplary structure of a peptide-linker compound. Fig. 2 shows an exemplary structure of a peptide-agent and a peptide-bonding agent-vehicle compound of the present invention. Figure 3 shows the nucleic acid and amino acid sequences of human IgG1 (SEQ ID NOS: 3 1 and 32, respectively) which can be used as a preferred vehicle in the present invention. Figure 4 shows exemplary Fc monomers and dimeric compounds of the invention, which may be derived from IgGl antibodies. "Fc" in the figure represents any F c variant that is consistent with the meaning of the Fc functional site herein. "Peptide" means any peptide, linker _ skin, skin-face combination or any combination thereof , as disclosed herein. Specific dimers are as follows: Figures 4A and 4D show a single disulfide bond dimer. 1 § (31 antibody typically has two disulfide bonds in the hinge region of the antibody. The FC functional sites in Figures 4A and 4D may be via truncation between two disulfide bond sites or via non-reactive residues (eg, propylamine) is formed by substituting a cysteamine thiol residue. The Fc functional site in Figure 4A is bonded to the amino terminus of the peptide; in Figure 4D, at the carboxy terminus of the peptide. Figure 4B and 4E shows a double disulfide-bonded dimer. This Fc functional site can be retained by truncating the parent antibody to retain two cysteamine thiol residues in the Fc functional site chain, or by encoding from these Fc functional sites. The structure of the sequence is formed. In Figure 4B, the Fc functional site is bonded to the amino terminus of the peptide; in Figure 4E, at the carboxy terminus of the peptide. Figures 4C and 4F show non-covalent dimerization. The Fc functional site can be formed by re-purchasing, or substituting, to eliminate cysteamine-based residues, as described in the invention, to eliminate cysteamine-based residues. Reactivity of a cysteamine thiol residue with a cysteamine thiol residue contained in other proteins in the host cell Impurity formed. Non-covalent bonding of the Fc functional site is sufficient to hold the dimer together. Other dimers can be formed using Fc functional sites derived from different types of antibodies (eg, IgG2, IgM). Figures 4G and 4H show the single-stranded Fc functional site bound to the N-terminus of the peptide (Figure 4G) and the c-terminus of the peptide (Figure 4H). Figure 5 shows an exemplary structure of the preferred compound of the present invention, It is characterized by a cascade of pharmaceutically active peptides that bind to an Fc functional site. Figure 5-8 shows that a single-stranded (or Fc monomer) molecule has a cascade of peptide dimers bound thereto, and also The DNA construct of the molecule is shown. Figure 5B shows that the Fc dimer 'where the linker-peptide moiety is only present on one bond of the Fc dimer. Figure 5C shows the peptide moiety on both strands Fc dimer (in this case 'is a cascade of peptide dimers.) The dimerization system of Figure 5C, in some host cells, after expression of a single stranded DNA construct as shown in Figure 5A Naturally formed. In other host cells, the cells can be placed in conditions suitable for forming a dimer, or the dimerization It can be formed in vitro. Figures 5A to 51 represent preferred examples of other exemplary single-stranded (F c monomers) and double-stranded (F c dimers). Figure 6 shows a preferred vector (20003180) The nucleotide sequence (SEQ ID NO 33) and the amino acid sequence (SEQ ID NO 34) are used to construct a TMP-Fc fusion compound, as shown in Example 3 herein. Figure 7 shows an exemplary oligomeric nuclear acid. Pair of fragments for better formation -18-016634 1316961 (14) Summary of the invention Continuation page 'The peptide of the present invention, as shown in Example 3. The nucleotide sequence and the amino acid sequence are respectively presented (SEQ ID NO 35-93). Figure 8 shows the nucleotide sequence (SEQ ID NO 94) and the amino acid sequence (SEQ ID NO 95) of the exemplary vector (20003 182) for constituting a C-terminal Fc fusion compound (ie, binding at the N-terminus) The C-terminal peptide of Fc). Figure 9 shows ELISA dose-response for selected phage colonies. Figures 10, 11 and 12 show the biological activity of selected compounds of the invention. Figures 13 and 14 show the _ count of platelets in vivo after a single injection of selected compounds of the invention in mice. Modes of Implementation I. Definition of Terms The terms used throughout this specification are defined as follows, unless otherwise limited in specific circumstances. The term ''peptide" refers to a molecule having from about 2 to about 80 amino acids, preferably from 3 to 40 amino acids. The exemplary peptides can be randomly generated by any of the methods described herein, such as Peptide library method (such as sputum display library), produced by chemical synthesis, derived from protein digestion or similar. The term "randomization" when used with a peptide sequence refers to a completely random sequence (such as the selection of autophages) The residue of the naturally occurring molecule of one or more of the sequence and the RNA-peptide screening method and the sequence is replaced by an amino acid residue which is not present at the position within the naturally occurring molecule. Exemplary methods for identifying randomized peptide sequences include phage display, E. coli display, ribosome display, RNA-peptide screening, chemical screening, and the like. -19- 016635 1316961 (15) The term "dimer" as used in the peptide refers to a molecule having two peptide chains, the two peptide chains being covalently or non-covalently bound, with or without a linking agent. The two peptides in the peptide dimer are C-terminal to N-terminally linked and may also be referred to as "cascade repeats" or π cascaded dimers. The two peptides in the peptide dimer are C- to C-terminal, or Ν-to-Ν-end linkage, and may also be referred to as "parallel repeats" or "parallel dimers". The term "polymer" as used in a peptide refers to a molecule having three or more peptide chains, which are covalent, non-covalent or both covalent and non-covalent. The covalent bond interacts with each other, with or without a linking agent. The term "derived π and π derivatives" or "derived" refers to the following methods and the resulting compounds, wherein (1) the compound is cyclic a moiety; such as cross-linking between cysteamine residues in a compound; (2) a compound cross-linked or having a cross-linking moiety; such as a compound having a cysteamine residue, thereby being in a medium or in vivo Forming a cross-linked dimer; (3) one or more pheno-peptide linkages are replaced by a non-peptidyl linkage; (4) Ν-terminal via -NRR1, NRCCCOR1, -NRCCCOOR1, -NRSCOhR1, - NHC(0)NHR, butyl quinone imine, or substituted or unsubstituted benzyloxycarbonyl-NH-, wherein R and R1 are ring substituents, as defined below; (5)C - the terminal is substituted with -C(0)R2 or -NR3R4, wherein R2, R3 and R4 are as defined below; and (6) the individual amino acid moiety of the compound is permeabilized The treatment of the agent reactive with the selected side chain or terminal residue is modified. The derivative is further described below. The term "platelet-forming analog peptide", "TPO mimetic peptide" or "TMP" a peptide which binds to the mpl receptor and/or which has platelet-producing activity -20-1316961 (16). It can also stimulate platelets or platelet precursors in vivo or in vitro, including but not The ability to produce megakaryocytes is limited. The term nmpl-binding functional site '' refers to any amino acid sequence that binds to the mpl receptor and includes naturally occurring sequences or randomized sequences. Example mpl binding function The site can be identified or derived by phage display or other methods described herein. The term nmpl receptor antagonist" refers to a molecule that binds to an mpl receptor and that increases or decreases one or more assay parameters, such as Source thrombopoietin (eTPO), a native mp} receptor ligand. The term π includes "includes" that a compound may include an additional amino acid at one or both of the Ν-terminus or C-terminus of a given sequence. Of course, these additional amino acids should not significantly interfere with the activity of the compound. Furthermore, physiologically acceptable salts of the compounds of the invention are also contemplated. The term "physiologically acceptable salt" refers to any salt known or later found to be pharmaceutically acceptable. Some specific examples are: acetate, trifluoroacetate, hydrohalide such as hydrochloride and hydrobromide. Acid salt, sulphate, citrate, tartrate, glycolate and oxalate. The term "vehicle" refers to a molecule that prevents degradation of therapeutic proteins and/or prolongs half-life and reduces It is toxic, reduces immunogenicity and/or increases biological activity. Exemplary vehicles include Fc functional sites (which are preferred) and linear polymers (such as polyethylene glycol (PEG), polylysine, dextran, etc.), branched polymers (see, for example, 1981) U.S. Patent No. 4,289,872 issued to Denkenwalter et al., issued Sep. 25, 1993; U.S. Patent No. 5,229,490 issued to Tam, on July 20, 1993, and issued on October 28, 1993 - 21 - 016637 1316961

WO 93/21259 by Frechet et al., lipids, cholesterol (such as steroids), saccharides or oligosaccharides (such as dextran), any natural or synthetic protein, multi-peptide that binds to salvage receptors Or peptide; albumin, including human serum albumin (HSA), leucine zipper domain, and other such proteins and protein fragments. The term "native Fc" refers to a molecule or sequence comprising a digested non-antigen-binding fragment sequence derived from the entire antibody, whether in monomeric or multimeric form. The original immunoglobulin source of native Fc is preferably of human origin and may be any immunoglobulin, although IgGl and IgG2 are preferred. The natural Fes is formed by a monomeric multipeptide which is bonded to a dimeric or multimeric form by a covalent bond (i.e., a disulfide bond) and a non-covalent bond. The number of intermolecular disulfide bonds between the haplotype subunits of the native Fc molecule ranges from 1 to 4, depending on the class (eg, IgG, IgA, IgE) or subclass (eg, IgG, IgG2, IgG3, IgAl, IgGA2) An example of a natural Fc is a disulfide-bonded dimer derived from a papain digestor of IgG (see Ellison et al. (1982), Nucleic Acids Res_ 10: 4071-9). The term "natural Fc" as used herein is a generic term for monomeric, dimeric, and multimeric forms. The term "Fc variant," refers to a molecule or sequence that is modified from a native Fc but still includes a binding site for a salvage receptor (FcRn). The international application, WO 97/3463 1 (published on September 25, 1997) and WO 96/32478, describe exemplary Fc variants and interactions with salvage receptors, which is incorporated herein by reference in its entirety. Thus the term "Fc variant" includes molecules or sequences derived from non-human native FC. In addition, native Fc includes removable sites because the structural properties or biological activities provided by them are not the fusion molecules of the present invention 016638 -22-1316961 (18). Thus, the term nFc variant "includes a molecule or sequence lacking one or more native Fc sites or residues that affect or involve (1) the formation of a disulfide bond, (2) with the host of choice Cell incompatibility, (3) N-terminal heterogeneity after expression in the host cell of choice, (4) glycosylation, (5) interaction with complement, and (6) binding to Fc receptor Rather than salvage receptors, or (7) antibody-dependent cellular cytotoxicity (ADCC). The term "Fc functional site" π includes the native Fc and Fc variant molecules and sequences as defined above. With respect to F c variants and native F cs, the term "F c functional site includes monomers in the form of monomers or poly-forms, whether digested from whole antibodies or formed by other methods. "Polymer" as used in the Fc functional site or Fc-containing functional site molecule, refers to a molecule having a multi-peptide chain with two covalent bonds or non-covalent bonds. The term "polymer" is used for Fc function. A site or a molecule comprising an Fc functional site refers to a molecule having two or more multi-peptide chains that interact by a covalent bond, a non-covalent bond, or a covalent bond and a non-covalent bond. IgG molecules typically form dimers; IgM, pentamer; IgD, dimer; and IgA, monomeric, dimeric, trimer or tetramer. The formation of the multimer can be achieved by using the sequence of the Fc source of Fc and the resulting activity or by derivatization (as defined herein) such a native Fc. The term "peptide antibody" or "peptide antibody population" refers to a molecule comprising a Fc functional site of an antibody that binds to at least one peptide. Such a peptide antibody can be a multimer or a dimer or a fragment thereof. It can be derived. II. Structure of the compound-23- 1316961 Jane Mi (19) In summary, the present invention proposes a compound which can bind and/or modulate the biological activity of the mpl receptor. More specifically, the present invention proposes a group. a compound which binds to the mpl receptor and/or transmits, ie, activates the mpl receptor to trigger a transmembrane signal that is identical to a receptor for the activity of the endogenous thrombopoietin (τρ〇). Thus, the compound of the invention has Platelet-producing activity, 'the ability to stimulate platelet production in vivo and in vitro and/or have megakaryocyte-forming activity', is the ability to stimulate platelet precursors, including megakaryocytes, in vivo and in vitro. In short, this - The inventive compound comprises one or more peptides of the formula I: X: X1-X2-X3-X4-GPTL-X9-X10-WL-X13-X14-X15-X16-X17-X18; Χ4, Χ9-Χ10 and χΐ3-Χ18 each In the composition of matter prepared according to the present invention, the compounds may comprise one or more peptides having the sequence of formula I, which are linked to each other or otherwise linked, Such as a dimer or a multimer. In another composition of matter prepared according to the present invention, the compounds may comprise one or more peptides of the formula I, which are at the Ν-end of the peptide or The c_ terminus is attached or otherwise linked to a vehicle. Any of these peptides may be cascaded (ie, sequentially, N to C), or parallel (ie, N-terminal to N-terminal' Or C-terminus to C-terminus, with or without a linking agent. Peptides. The compounds of the invention include indole analog peptides, either alone or in combination with other TMPs, such as 'dimers or polymers. The tmps of the present invention include the following sequences: I : X1-X2-X3-X4-GPTL-X9-X10-WL-X13-X14-X15-X16-X17-X18; -24· 016640 1316961 (20) Occupation: Wherein X1-X4, X9-X10 and X13-X18 are each independently an amino acid. Preferred amino acid residues of the above sequence are further defined in Table 1 below. Preferred amino acid residues 1-

Position amino acid residue XI A'V'W'M'G'Y'C'Q'E'R'H X2 A, V, L, I, G, S, C X3 L, I, P, W , G, S, D, K, R X4 L, G, Q, D, E, Η X9 K, R. X10 Q, E X13 A, V, L, S, Q, E, R X14 A, W, T, Y, C, Q X15 V, L, G, Y, R X16 A, L, F, G, R X17 A, V, L, M, G, C, Q, N X18 A, V, P, M, F, G, C, Q, K and even better TMP sequences of the invention are peptides having the following sequence: I : X1-X2-X3-X4-GPTL-X9-X10-WL-X13-X14-X15 -X16-X17-X18; wherein X1-X4, X9-X10 and X13-X18 are each independently an amino acid, and wherein the peptide has binding affinity to the mpl receptor, and/or has the following sequence The same or greater biological activity: IEGPTLRQWLAARA [SEQ ID NO 1]. Binding affinity can be measured by any assay available or known to the skilled artisan, including but not limited to BIAcore measurement, ELISA assay, competition -25- 0'6641 1316961, invention, 'continued page, ^ __________ (21 ) Sexual tests, etc. Biological activity can be measured in vivo or in vitro by any assay available or known to the skilled artisan. Exemplary assays include, but are not limited to, cell-based assays, ie, megakaryocyte proliferation assays, 32D cell assays (IL-3 dependent selection of murine 32D cells transfected with human mpl receptors, as detailed in WO 95/26746) Medium), CD34+ assay, CD61 cell assay, etc. Biological activity can also be measured by various in vivo animal assays.

The preferred TMP sequences of the invention are identified in Table 2 below. Table 2 - Sequence preferred TMP TMP No. PEPTIDE SEQUENCE SEQ ID NO: TMP2 GAREGPTLRQWLEWVRVG 2 TMP3 RDLDGPTLRQWLPLPSVQ 3 TMP4 ALRDGPTLKQWLEYRRQA 4 TMP 5 ARQEGPTLKEWLFWVRMG 5 TMP6 EALLGPTLREWLAWRRAQ 6 TMP7 MARDGPTLREWLRTYRMM 7 TMP8 WMPEGPTLKQWLFHGRGQ 8 TMP9 HIREGPTLRQWLVALRMV 9 TMP 10 QLGHGPTLRQWLSWYRGM 10 TMP 11 ELRQGPTLHEWLQHLASK 11 TMP 12 VGIEGPTLRQWLAQRLNP 12 TMP 13 WSRDGPTLREWLAWRAVG 13 TMP 14 AVPQGPTLKQWLLWRRCA 14 TMP 15 RIREGPTLKEWLAQRRGF 15

-26- 016642 1316961 ΤΜΡ16 RFAEGPTLREWLEQRKLV 16 ΤΜΡ17 DRFQGPTLREWLAAIRSV 17 ΤΜΡ18 AGREGPTLREWLNMRVWQ 18 ΤΜΡ19 ALQEGPTLRQWLGWGQWG 19 ΤΜΡ20 YCDEGPTLKQWLVCLGLQ 20 ΤΜΡ21 WCKEGPTLREWLRWGFLC 21 ΤΜΡ22 CSSGGPTLREWLQCRRMQ 22 ΤΜΡ23 CSWGGPTLKQWLQCVRAK 23 ΤΜΡ24 CQLGGPTLREWLACRLGA 24 ΤΜΡ25 CWEGGPTLKEWLQCLVER 25 ΤΜΡ26 CRGGGPTLHQWLSCFRWQ 26 ΤΜΡ27 CRDGGPTLRQWLACLQQK 27 ΤΜΡ28 ELRSGPTLKEWLVWRLAQ 28 ΤΜΡ29 GCRSGPTLREWLACREVQ 29 ΤΜΡ30 TCEQGPTLRQWLLCRQGR 30 DESCRIPTION OF THE INVENTION The binding affinity and biological activity data of the contiguous peptide TMP2-TMP30 are further illustrated in the Examples. In order to better mimic the phage environment used to select the peptide, and to mask the charged amine and carboxy termini of the preferred 18 amino acid peptide, add two amines to each end of each peptide. Acidic ''cover'. In particular, glutamic acid (Q) and cysteine (C) were added to the amine end of each TMP2-TMP30. Similarly, two amino acids were added to the carboxy terminus of each peptide to cover π--histamine (H) and serine (S). It is understood by those skilled in the art that such caps are only used to mask the charged ends and are not intended to increase or decrease the binding affinity and/or biological activity of the preferred peptides. 016643 -27- 1316961 , _ (23) Description of the invention continued, page words., £Λκ k ^ιίλ 1&ι_λ<^Ι!1ι^·1ί{ίίΰτ·ΰ Since the affinity of the peptide is known to vary with the length of the peptide In addition, the reference bioactive peptide (SEQ ID NO 1) was increased from 14 amino acids to 22 amino acids to have the same length as the test peptide TMP2-TMP30. See Examples 6-11.于此 It is understood by those skilled in the art that the 14 amino acid sequences of the bioactive region of the peptide are compared, as identified in SEQ ID NO 1, and are also referred to as TMP1. Any peptide containing a cysteamine thiol residue can be cross-linked to another Cys-containing peptide, either or both of which can be linked to a vehicle. Any peptide containing more than one Cys residue can also form a disulfide bond within the peptide. Any of these peptides can be derived as described below. Other useful peptide sequences are derived from the conservative and/or non-conservative modification of the amino acid sequences of the TMPs disclosed herein. Conservative modifications result in peptides with functional and chemical characteristics similar to those of the peptides that perform such modifications. In contrast, substantial modification of the functional and/or chemical characteristics of the peptide can be accomplished by selective substitution in the amino acid sequence such that they differ significantly in their effectiveness in maintaining the following characteristics (a) The structure of the main molecular skeleton in the substitution zone, such as a sheet or helical configuration, (b) the charge or hydrophobicity of the molecule at the target site, or (c) the size of the molecule. For example, a "conservative amino acid substitution" can include the substitution of a natural amino acid residue with a non-native residue such that there is little or no effect on the polarity or charge of the amino acid residue at that position. Any natural residue in the multi-peptide may also be substituted by alanine as previously described in "Alanine Scanning Mutation Formation" (see, eg, MacLennan et al., 1998, Acta Physiol. Scand. Suppl. 643: 55- 67; Sasaki et al., 1998, Adv. Biophys. 35:1-24, which discusses the formation of alanine scanning mutations.) -28- 016644 1316961 (24) The invention describes the amino acid substitution required for the continuation of the page ( Whether conservative or non-conservative, it is up to the skilled person to decide when such substitutions are required. For example, amino acid substitutions can be used to identify important residues of the peptide sequence, or to increase or decrease the Affinity of the peptide or vehicle-peptide peptide (see the previous formula). Exemplary amino acid substitutions are listed in Table 3. Table 3 - Amino acid substitution of the original residue Example substitution of the preferred substitution Ala (A) Val, Leu, lie Val Arg (R) - Lys, Gin, Asn Ly s Asn (N) Gin Gin Asp (D) Glu Glu Cys (C) Ser, Ala Ser Gin (〇) Asn Asn Glu (E) Asp Asp Gly (G) Pro, Ala Ala His (H) Asn, Gin, Lys , Arg Arg lie(1) Leu, Val, Met, Ala, Phe, Leuk Leu (L), leucine, lie, Val, Met, Ala, Phe lie Lys (K) Arg, 1,4-diamine Butyric acid, Gin, Asn Arg Met (M) Leu, Phe, lie Leu Phe (F) Leu, Val, lie, Ala, Tyr Leu Pro (P) Ala Gly Ser (S) Thr, Ala, Cys Thr Thr ( T) Ser Ser -29- 016645 1316961

Trp(W) _ Tyr ' Phe Tyr(7) Trp, Phe, Thr, Ser Val (V) lie, Met, Leu, Phe, Ala, orthanoic acid

Leu In certain embodiments, conservative amino acid substitutions can also include >~1" encompasses non-naturally occurring amino acid residues, which typically pass through the chemical system, rather than through the biological system. The synthesis is incorporated. Naturally occurring residues can be classified into several classes based on common side chain properties that can be used for sequence eve & μ ^, 々 < For example, a non-conservative substitution may include the exchange of one of the members of this class with one of the other classes. Such substitution residues can lead to a peptide region homologous to a non-human ortho log' or to a non-homologous region of the molecule. In addition, P or G can also be used for modification to achieve the purpose of influencing the orientation of the bond. For such modifications, the hydropathic index of the amino acid can be considered. Each amino acid specifies a hydrophobic index based on its hydrophobicity and charge characteristics, which are: isoleucine (+4.5); proline (+4.2); leucine (+3.8); phenylalanine (+) 2.8); cysteine/cystine (+2.5); methionine (+1.9); alanine (+1.8); glycine (-0.4); threonine (_〇7); silk Aminic acid (-0.8); tryptophan (-0.9); tyrosine (-1.3); valine (4.6); histidine (-3.2); glutamic acid (-3.5); glutamine (-3.5); aspartic acid (-3.5); aspartame (-3.5); lysine (-3.9); and arginine (_45). The importance of hydrophobic amino acid indices in making proteins biologically interactive is known to those skilled in the art. Kyte et al., J. Mol. Biol., 1 57: 105- 13 1 (1982). It is known that certain amino acids can replace other amino acids having similar hydrophobic indices or values while still maintaining similar biological activities.于基-30-1316961 (26) Description of the Invention In the change in the hydrophobic index, the amino acid substitution between the hydrophobic index and the soil 2 is preferred, and between ±1 is preferred. The best between ±0.5 or even smaller. It is also understood in the art that substitutions similar to amino acids can be effectively accomplished based on hydrophilicity. The maximum local average hydrophilicity of a protein, as controlled by the hydrophilicity of its adjacent amino acids, is related to its immunogenicity and antigenicity, i.e., to the biological properties of the protein. The following hydrophilicity values are assigned to the amino acid residues: spermine (+ 3.0); lysine (+ 3_0); aspartic acid (+ 3.0 ± 1); glutamic acid (+ 3.0 ± 1) ; serine ( + 0.3); aspartame ( + 0.2); glutamic acid ( + 0.2); glycine (0); threonate (-0.4); proline (-0 · 5) ±1); alanine (-0.5); histidine (-0.5); cysteine (-1.0); methionine (-1.3); valine (-1.5); leucine (- 1.8);isoleucine (-1.8); tyrosine (-2.3); phenylalanine (-2.5); tryptoic acid (-3.4). Among the changes based on similar hydrophilicity values, it is preferred that the amino acid is substituted with a hydrophilicity value of between ± 2, preferably between ± 1 and between ± 0.5 or even more. Small is the best. The epitope can also be identified from the original amino acid sequence based on hydrophilicity. These regions are also referred to as "antigenic core regions". Those skilled in the art can use known techniques to determine appropriate variants. In order to identify suitable regions in the molecule that can be altered without disrupting activity, those skilled in the art can target regions that are believed to be unimportant to activity. For example, when a similar multi-peptide having the same activity from the same species or a different species is known, the skilled person can compare the amino acid sequence of the peptide with the similar peptide. In this comparison, residues that are conserved relative to similar polypeptides in the molecule can be identified. 016647 - 31 - 1316961 (27) Illustrated on page continuation. It will be appreciated that changes in the region of the peptide that are not conserved relative to such similar peptides are less likely to negatively affect the biological activity and/or structure of the peptide. It will also be appreciated by those skilled in the art that even within a relatively conserved region, it is possible to replace naturally occurring residues with chemically similar amino acids while retaining activity (conservative amino acid residue substitution). Thus, conserved amino acid substitutions can be made even for biologically active or structurally important regions without disrupting biological activity or negatively affecting the peptide structure. The amino acid has L or D stereochemistry (other than Gly, which is neither L nor D), and the TMPs of the present invention may comprise a stereochemical combination. However, L stereochemistry is preferred for all amino acids in the TMP chain. The invention also provides a reverse TMP molecule wherein the amino- to carboxy-terminal sequence of the amino acid is reversed. For example, a molecule with a normal X^Xz-Xs sequence will have the opposite. The present invention also proposes a reverse-reverse TMP molecule in which, like reverse TMP, the amino- to carboxy-terminal sequence of the amino acid is reversed, and the residue of the usual "L" enantiomer in TMP is changed to nDn stereoisomeric forms. Derivatives of TMP are also contemplated to be substituted for the above TMP. Such TMP derivatives include those in which one or more of the following modifications can be made: • One or more peptides [-C (0) NR-] linkage is substituted by non-peptidyl linkage, such as -CH2-carbamate linkage [-CH2-0C(0) NR-]; phosphate linkage; -CH2- Sulfonamide [-CH2-S(0)2NR-] linkage; urea [-NHC(0)NH-] linkage; -CH2-secondary amine linkage; or alkylation-peptidyl linkage [-C( 0) NR6- wherein R6 is lower alkyl]; • peptide, wherein N-terminal is derivatized to -NRR1 group; -NRC(0)R group; -32-016648 1316961 (28) Description of the Invention Continued - NRC (0)0R group; -NRS(0)2R group; -NHC(0)NHR group, wherein R and R1 are a hydrogen atom or a lower alkyl group, but the limitation is that R and R1 are not all hydrogen atoms; Butyl imino group; phenyl hydroxycarbonyl-NH-(CBZ-NH-) group; or phenyl hydroxycarbonyl-NH- group, having 1 to 3 options on the benzene ring The substituents in the following groups are included: a low carbon number base, a low number of alkoxy groups, chlorine and a desert; and a peptide, wherein the free C-terminus is derivatized to -C(0)R2, wherein R2 is as follows Selected from the group: a lower alkoxy group and -NR3R4, wherein R3 and R4 are each independently selected from the group consisting of a hydrogen atom and a lower alkyl group. "Low carbon number" means that the group has In addition, the modification of each amino acid can be conducted into the TMP molecule by reacting the target amino acid residue of the peptide with an organic derivatizing agent, and the derivatizing agent can be combined with the selected side chain. Or the terminal residue reaction. The following are examples: The amidoxime group and the amine terminal residue can be reacted with amber or other carboxylic anhydride. The derivatization of these agents has the effect of reversing the charge of the amine sulfhydryl residue. Suitable reactants for derivatization of the α-amino group-containing residue include sulfhydryl esters such as mercaptopyridinium imide; pyridoxal phosphate; pyridoxal; chloroborohydride; trinitrobenzenesulfonic acid; 0-methylisourea; 2,4-pentanedione; and transaminase catalyzed reaction with acetyl ester. Modification by reaction with one or more conventional reactants, including phenylglyoxal, 2,3-butanedione, 1,2-cyclohexanedione and ninhydrin. Derivatization of arginine residues The effect is due to the high pKa of the hydrazine functional group -33- 016649 1316961 (29) The reaction needs to be carried out under basic conditions. In addition, these reactants can also react with the amide group and the arginine group. A number of studies have been conducted on the specific modification of the ruthenium-based residue, with particular attention to the introduction of a spectrally labeled tyrosine-based residue by reaction with an aromatic diazonium compound or tetranitrononane. Most commonly, N-acetamimid and tetranitromethane can be used to form the 0-acetamidinoguanine species and the 3-nitro derivative, respectively. The pendant carboxyl group (aspartame or glutamine) can be selectively modified by reaction with carbodiimide (R'-N=C = NR·), such as 1-cyclohexyl-3- (2-morpholinyl-4-ethyl)carbodiimide or 1-ethyl-3-(4-azo-4,4-dimethylpentyl)carbodiimide. In addition, aspartame and glutamine residues can be converted to aspartame and glutamine residues by reaction with ammonium ions. The glutamine and aspartame amide groups typically remove the guanamine group to the corresponding glutamine S& base and aspartame stabilizing groups. Alternatively, these residues can be removed from the guanamine group under mildly acidic conditions. Any form of these residues is within the scope of the invention. Derivatization using a difunctional reactant can be used to crosslink the peptide or its functional derivative to a water insoluble support matrix or other macromolecular carrier. Commonly used crosslinking agents include, for example, 1,1-bis(disazoethyl)-2-phenylethane, glutaraldehyde, N-hydroxybutylimine, such as with 4-azido An ester of the salicylic acid, a homobifunctional quinone imide, including a diammonium imino ester such as 3,3'-dithiobis(succinimide propionate) and a bifunctional Malay Indole imines, such as bis-N-malekulimino-1,8-octane. Derivatizing agents such as methyl-3-[(p-azide-34-016650 1316961 (30) invention continuation-phenyl)dithio]propanoid imidate can form a cross in the presence of light A photoactivated intermediate bonded to a bond. In addition, reactive water-insoluble matrices such as cyanogen bromide-activated saccharides and reactive matrices described in the following documents can be used for protein immobilization: U.S. Patent Nos. 3,969,287; 3,691,016; 4,195,128; 4,247,642; 4,229,537 and 4,330,440 Other possible modifications include carboxylation of valine and lysine, phosphorylation of hydroxy groups in serine sulfhydryl or sulphalidyl residues, oxidation of sulfur atoms in cysteine, lysine, spermine Methylation of α-amino groups in acid and histidine side chains (Creighton, TE, Proteins: Structure and Molecule Properties, WH Freeman & Co_, San Francisco, pp. 79-86 (1983)), N-terminal The acetylation of the amine and, in some instances, the amide amination of the C-terminal carboxyl group. Such derived fractions preferably improve one or more of the properties of the compounds of the invention, including platelet production activity, solubility, absorbency, biolife, and the like. In addition, the derivatized portion may result in a compound having the same or substantially the same properties and/or properties as the underivatized compound. This moiety may additionally eliminate or impair the adverse effects of the compound and the like. The compounds of the invention may also be altered at the DNA level. The DNA sequence of any portion of the compound may be changed to a more compatible codon to the host cell of choice. For better host E. coli, the optimized codon system is known in the art. The codon can be substituted to eliminate restriction sites or include a silent restriction site that facilitates DNA processing within the selected host cell. The DNA sequences of the vehicle, the linker and the peptide can be modified to include any of the foregoing sequence changes. Therefore, the scope of the invention is discussed in the above-mentioned paragraph - 35- 016651 1316961 (31) Description of the invention; some modifications, substitutions, derivations, etc., apply to all aspects of the invention, including but not limited to peptides, peptide dimers And polymers, bonding agents and vehicles. In addition, the skilled artisan can review structure-function studies to identify residues that are important for activity and structure in similar peptides. In view of such information, the skilled person can predict the importance of amino acid residues in the peptide corresponding to amino acids which are important for activity or structure in similar peptides. Alternatively, a chemically similar amino acid may be substituted for such predicted important amino acid residues in the peptide. It is also possible for the skilled artisan to analyze the three-dimensional structure and the amino acid sequence in relation to the structure of a similar multi-peptide. With this information in mind, those skilled in the art may choose not to make fundamental changes to the amino acid residues expected to appear on the surface of the protein' because such residues may involve important interactions with other molecules. In addition, one skilled in the art can produce test variants which contain a single amino acid substitution in each of the desired amino acid residues. The variants are then screened using activity assays known to those skilled in the art. Such data can be used to collect shells for appropriate variants. For example, variants with such changes should be avoided if changes in one-to-one specific amino acid residues are found to result in disruption of activity, undesired reduction or inappropriate activity. In other words, based on the information gathered from such routine experiments, the skilled artisan can readily determine which amino acids should be avoided, either alone or in combination with other mutations. A large number of scientific articles are devoted to the prediction of secondary structure. See Moult J., Curr. Op. in Biotech., 7(4): 422-427 (1996), Chou et al., Biochemistry, 1 3(2): 222-245 (1 974); Chou et al. , Biochemistry, 113 (2): 211-222 (1974); Chou et al., Adv. Enzymol. Relat. Areas Mol. -36- 016652 1316961 (32)

Biol., 47* 45-148 (1978) j Chou et al. 3 Ann. Rev. Biochein., 47i 251-276 and Chou et al_, Biophys. J., 26:367-384 (1979). In addition, existing computer programs can help predict secondary structure. One method of predicting secondary structure is based on homology modeling. For example, two multi-peptides or proteins, which have a sequence identity of more than 30%, or a similarity of more than 40%, often have similar structural topologies. The recent growth of the Protein Structure Database (PDB) has led to the predictability of secondary structure enhancement, including the number of possible folds in multipeptide or protein structures. See Holm et al., Nucl. Acid. Res., 27(1): 244-247 (1999). Brenner et al., Curr· Op. Struct·Biol·, 7(3): 369-376 (1997) speculates that there is a finite number of folds in a given multi-peptide or protein, and once the critical number of structures is resolved , you can get very accurate structural predictions. Other methods for predicting secondary structure include "threading" (Jones, D., Curr. Opin. Struct. Biol., 7(3): 377-87 (1997); Sippl et al., Structure, 4(1): 15-9 (1996)), "profile analysis" (Bowie et al., Science, 253: 164-170 (1991); Gribskov et al., Meth. Enzym. , 183:146-159 (1990); Gribskov et al., Proc. Nat. Acad. Sci., 84(13): 4355-8 (1987)), and "Evolutionary Linkage,, (see Home, supra, And Brenner, supra) 0 The general formula of the preferred peptide and peptide-bonding agent molecules of the present invention is shown in FIG. In addition, physiologically acceptable salts of TMPs are also contemplated. Peptide Compounds In addition to novel peptides, the present invention provides novel peptide compounds in which one or more of the peptides of the present invention are followed by a linking agent (LN) and/or a vehicle (V) -37-016653 1316961 (33) Description of the invention contiguous; on, or otherwise linked to each other. The TMP can be cascaded (i.e., sequentially, N- to C-terminus) or parallelly linked (i.e., N- to N-terminus or C- to C-terminus). The TMP can be followed by other TMPs or the same TMP with or without a linker. The TMP can also be followed by other TMPs or the same TMP with or without a linking agent and with or without a vehicle. The peptide-bonding agent-vehicle compound of the present invention can be illustrated by the following formula;

II (Vl)v--(LNl)1--(TMPl)a--(LN2)m--(TMP2)b--(LN3)n--(TMP3)c- wherein VI and V2 are vehicles; LN1, LN2, LN3 and LN4 are each independently a linking agent; TMP1, TMP2, TMP3 and TMP4 are each independently a peptide sequence of formula I; a, b, c and d and 1, m, η And 〇 are each independently an integer from 0 to 20, and ν and w are each independently an integer from 0 to 1. The compounds of the present invention are, for example, represented by the following formula: TMP1-V1 TMP1-LN1-V1 TMP1-TMP2-V1 TMP1-LN1-TMP2-LN2-V1 and other multimers thereof, wherein VI is a vehicle (cf. The best is the Fc functional part), and the linkage is at the C-terminus of TMP, with or without a linking agent; V1-TMP1 V1-LN1-TMP1 V1-TMP1-TMP2 V1-LN1-TMP1-LN2-TMP2 and These other multimers, wherein VI is a vehicle (preferably an Fc functional site), and are linked at the N-terminus of TMP, with or without a linking agent. The general formula of the preferred peptide-vehicle and peptide-bonding agent-vehicle molecule of the present invention is shown in Fig. 38-016654 1316961 (34). Many of the preferred compounds of the invention are dimers or polymers in which they have two TMP moieties, or multimers, which have multiple TMP moieties. Each of TMP1 to TMP4 and the like may have the same or different structure. Preferably, the compounds of the invention have from 2 to 5 TMP moieties, particularly preferably from 2 to 3, and most preferably two. These compounds are preferably dimers which are bound directly or via a linkage (see below). The monomeric TMP moiety system is shown in a conventional orientation, read from left to right. Take the N-to-C-end solution. Accordingly, it can be seen that the compounds of the present invention can be oriented such that the C-terminus of TMP1 binds directly or through a linkage to the N-terminus of the TMP2 (cascade dimer). In addition, the compound of the present invention may be oriented such that the C-terminus of TMP1 is directly bonded to or bonded to the C-terminus of TMP2 via a linkage, or the N-terminus of TMP1 is directly bonded to or through a linkage to TMP2. N-terminal (parallel dimer). These compounds are called dimers, even though TMP1 and TMP2 are structurally distinct. That is, it can be constructed as a homodimer and a heterodimer. Bonding agent In another embodiment, the invention proposes that one or more TMPs are covalently bonded via a ''linking agent' group (LN1, LN2, etc.) or otherwise linked, or combined with another On TMP, any linkage group is optional. When it is present, its chemical structure is not important because it acts primarily as a spacer. The linkage should be chosen so that it does not affect the biological activity of the final compound. It is also possible to make the immunogenicity of the final compound not significantly improved. The bonding agent is preferably composed of an amino acid which is bonded through a peptide bond. Therefore, it is preferably in the form of -39-016655 1316961 (35) In the contingent embodiment, the linking agent consists of from 1 to 30 amino acids bonded by a peptide bond selected from the group consisting of 20 naturally occurring amino acids. Some of these amino acids Glycosylation, as is well understood by those skilled in the art. In a more preferred embodiment, from 1 to 20 amino acids are selected from the group consisting of glycine, alanine, proline, and aspartic acid. Amine, glutamine, and lysine. More preferably, the bonder is composed of a main body of an amino acid. It is not hindered, such as glycine and alanine. Therefore, the preferred linking agent is polyglycine (especially (Gly) 4, (Gly) 5), poly (Gly-Ala) and polypropylamine. Specific examples of other linkage agents are: (Gly)3Lys(Gly)4 (SEQ ID NO: 96); (Gly)3AsnGlySer(Gly)2 (SEQ ID NO: 97); (Gly)3Cys(Gly) 4 (SEQ ID NO: 98); and GlyProAsnGlyGly (SEQ ID NO: 99) To explain the above terms, such as (Gly)3Lys(Gly)4 represents Gly-Gly-Gly-Lys-Gly-Gly-Gly-Gly-.Gly It is also preferred to use it with Ala. The linking agents shown herein are exemplary; the linking agents within the scope of the invention may be longer and may include other residues. Non-peptide-bonding agents are also possible. For example, an alkyl linkage agent such as -NH-(CH2)sC(0)-, wherein s=2-20 can be used. These alkyl linkage agents can be further substituted by any non-steric hindrance group, such as a lower alkyl group. (eg C^-Ce) low sulfhydryl, halogen (such as chlorine, bromine), CN, NH2, phenyl, etc. An example of a non-peptide-bonding agent is a polyethylene glycol linkage, 016656 -40- Invention T-page · 1316961 (36)

Η wherein η is such that the bond has a molecular weight of from 100 to 5,000 kD, preferably from 100 to 500 kD. The peptide-bonding agent can be altered to form a derivative in the manner described above. In general, a linking agent having a length of about 0 to 14 units (e.g., an amino acid) has been found to be preferred for the platelet-forming compound of the present invention. The peptide-bonding agent can be modified in the same manner as described in the above TMP to form a derivative. Further, the compound of this specific example may further be linear or cyclic. "Circular" means that at least two separate, i.e., non-contiguous, portions of the molecule are bonded to each other. For example, the amino terminus and the carboxy terminus of the terminal of the molecule may be covalently bonded to form a cyclomeric molecule. Alternatively, the molecule may comprise two or more Cys residues (as in a linking agent) which are cyclizable by the formation of a disulfide bond. It is further contemplated that more than one cascade of peptide dimers can be linked to form a dimer dimer. Thus, for example, a cascaded dimer containing a Cys residue can form an intermolecular disulfide bond with the Cys of another such dimer. EXAMPLES The peptide-bonding compound of the present invention is as follows; CSSGGPTLREWLQCRRMQ--GGGGG--CSSGGPTLREWLQCRRMQ (SEQ ID NO 100); QLGHGPTLRQWLSWYRGM--(Gly)3Lys(Gly)4--ALRDGPTLKQWL EYRRQA (SEQ ID NO 101 );

RFAEGPTLREWLEQRKLV-GGG(PEG)GGG-RFAEGPTLREWLEQR -41 · 01G657 1316961 (37) KLV (SEQ ID NO 102). Accordingly, in a preferred embodiment, the linking agent comprises (LN1)n, wherein LN1 is a naturally occurring amino acid or a stereoisomer thereof, and "n" is any one of 1 to 20. The formula of the preferred peptide-linker molecule is shown in Figure 1. More preferred skin-linking agent molecules include: i) TMP1-LN1-TMP2-LN2 ii) LN1-TMP1-LN2-TMP2 iii) LN1-TMP1-LN2-TMP1 iv) TMP1-LN1-TMP1-LN1-TMP1 -LN1 v) LN1-TMP1-LN2-TMP2-LN3-TMP3-LN4-TMP4 wherein LN1-LN4 are each independently a linking agent. Vehicle In another embodiment, the peptide or peptide compound of the invention may be linked or subsequently attached to a vehicle (V). A vehicle generally refers to a molecule that prevents the degradation of therapeutic proteins and/or prolongs their half-life, reduces their toxicity, reduces immunogenicity, or increases biological activity. The vehicle (V) can be passed over the peptide via the N-terminus, the C-terminus, the peptide backbone or the side chain. The vehicle (V) can be a carrier molecule, such as a linear polymer (such as polyethylene glycol, polylysine, dextran, etc.), a branched polymer (see, for example, issued on September 15, 1981) U.S. Patent No. 4,289,872 to Denkenwalter et al.; U.S. Patent No. 5,229,490 issued to Tam, on July 20, 1993; WO 93/21259 to Frechet et al., issued Oct. 28, 1993; a base (such as a steroid); or a saccharide or oligosaccharide. Other possible vehicles include one or more water-soluble polymer addenda such as polyoxygen 016658-42- hair » Ming Xia: 1316961 (38) ethylene glycol, or polypropylene glycol, such as U.S. Patent No. 4,640,835 , 4,496,689, 4,301,144, 4,670,417, 4,791,192 and 4,179,337. Still other useful polymers known in the art include monomethoxy-polyethylene glycol, dextran, cellulose, or other sugar-based polymers, poly-(N-ethylheptyl p-pyrene Slave S)) - Polyethylene - drunk, propylene dihydrogen homopolymer, polyemulsified propionate / ethylene oxide copolymer, polyoxyethylated polyol (such as glycerol) and polyethylene glycol, and a mixture of these polymers . Exemplary vehicles also include: • an Fc functional site; • other proteins, polypeptides or peptides that bind to salvage receptors; • human serum globulin (HSA); • leucine zipper (LZ) domain; • Polyethylene glycol (PEG), including 5 kD, 20 kD and 30 kD PEG, and other polymers; • Glucan; and other molecules known in the art to provide extended half-life and/or avoidance Proteolytic degradation or clearance. An exemplary vehicle is polyethylene glycol (PEG). The PEG family can have any convenient molecular weight' and can be linear or branched. The average molecular weight of pEG is preferably from about 2 kDa to about 1 〇〇 kDa, more preferably from about 5 kDa to about 5 〇 kDa, and most preferably from about 5 kDa to about 10 kDa. The PEG family is typically passed through the following reactions followed by a compound of the invention: deuteration reductive alkylation, Micha^ii formation, thiol alkylation or other chemoselective conjugation/linkage, through a portion of the pEG moiety Reaction f-based group (such as aldehyde group, amine group, ester group, thiol, -haloethylidene group, Malay 醯 016659 1316961 (39) Description of the invention (Continuation 1 of the imine group or fluorenyl group) on the target compound The reaction group (e.g., aldehyde group, amine group, acetoxy group, thiol group, -haloethyl group, maleimide group or fluorenyl group) is reacted. The saccharide (oligosaccharide) can conveniently be attached to a glycosylation site known in the protein. In general, the 0-linked oligosaccharide is attached to a serine (Ser) or threonine (Thr) residue, and the N-linked oligosaccharide is followed by an aspartame. On the amine (Asn) residues, they are part of the Asn-X-Ser/Thr sequence, wherein X can be any amino acid other than proline. X is preferably one of the 19 naturally occurring amine base acids, but does not include proline. The N-linked and 0-linked oligosaccharides and the sugar residues found in each type are structurally different. One type of sugar that is commonly found in both is N-ethyl thio-neuraminic acid (referred to as sialic acid). The sialic acid is often the terminal residue of the N-linked and 0-linked oligosaccharides, and the glycosylated compound is made acidic by the negative charge it carries. Such a moiety may be added to the linking agent of the present invention, and is preferably glycosylated by the cell during recombinant formation of the multi-peptide compound (e.g., in mammalian cells such as CH0, BHK, COS). However, such sites can be further glycosylated by synthetic or semi-synthetic procedures known in the art. In a more preferred embodiment, the vehicle (V) may comprise one or more antibody Fc functional sites. Thus, the above peptide compounds can be further fused to one or more Fc functional sites, either directly or via a linkage agent. The Fc vehicle can be selected from the human immunoglobulin IgG-Ι heavy chain, see Ellison, JW et al., Nucleic Acids Res. 10: 4071-4079 (1982), or any other Fc sequence known in the art (eg other IgG family, including but not limited to IgG-2, IgG-3 and IgG-4, or other immunoglobulins). 016660 -44· 1316961 (40) Description of the Invention Continued! It is well known that the Fc functional part of an antibody is a monomeric multi-peptide synthesized by disulfide or non-covalent bond association into a dimeric or multimeric form. The segment is composed of segments. The number of intermolecular disulfide bonds between the haplotype subunits of the native Fc molecule ranges from 1 to 4, depending on the class of antibody (eg, IgG, IgA, IgE), or subclass (eg, IgGl, IgG2, IgG3, IgAl, IgGA2). The term "Fc" as used herein is generally referred to as the monomeric, dimeric, and multimeric forms of the Fc molecule. It should be noted that the Fc monomer can spontaneously dimerize when it contains a suitable Cys residue unless special conditions are present to prevent dimerization by the formation of a disulfide bond. Even if a Cys residue which normally forms a disulfide bond in the Fc-dimer is removed or substituted by another residue, the monomer chain usually dimerizes through non-covalent bond interaction. The term "Fc11 is used herein to mean any such form: natural monomers, natural dimers (disulfide bonds), modified dimers (disulfide and/or non-covalent bonds) And modified monomers (ie derivatives).

Variants, analogs or derivatives of the Fc moiety can be constructed by, for example, substituting various residues or sequences. Variant (or analog) polypeptides include intervening variants in which one or more amino acid residues complement the Fc amino acid sequence. The insertion can be at either or both ends of the protein or can be placed in the internal region of the Fc amino acid sequence. Insertion variants with added residues at either or both ends include, for example, fusion proteins and amino acid-containing labels or labeled proteins. For example, an Fc molecule may optionally comprise an N-terminal Met, particularly when the molecular recombination is manifested in a bacterial cell, such as E. coli. In the Fc deletion variant, one or more amino acid residues in the Fc polypeptide are deleted. Deletion can be performed on one or both ends of the Fc polypeptide, or deleted at -45-1316961 (41)

One or more residues within the Fc amino acid sequence. Therefore, the deletion variant includes all fragments of the F c multi-peptide sequence. In the Fc substitution variant, one or more amino acid residues of the Fc polypeptide are removed and replaced with an alternative residue. In one aspect, the substitution is conservative in nature, but the invention also encompasses non-conservative substitutions. For example, the 'half-transamine residue can be deleted or substituted with other amino acids to avoid some or all of the disulfide linkages that form the Fc sequence. Each of these semi-proline residues may be removed or substituted with one or more such cysteine residues with other amino acids such as Ala or Ser. As another example, modifications may be made to introduce an amino acid substitution to remove (受体) the receptor binding site; (2) to remove the complement (Clq) binding site; and/or (3) to remove antibody dependence Cytotoxicity (ADCC) site of cellular mediators. Such sites are known in the art, and any known substitutions are within the scope of the Fc used herein. See, for example, Molecular Immunology, Vol. 29, No. 5, 633-639 (1992) for the ADCC site in IgG1. Similarly, one or more tyrosine residues may also be substituted with amphetamine residues. In addition, the insertion, deletion (e.g., from 1-25 amino acid) and/or substitution of other variant amino acids are also contemplated and are within the scope of the invention. A conservative amino acid substitution is generally preferred. In addition, the modifications may be in the form of a modified amino acid, such as a peptide mimetic or a D-amino acid. The Fc sequences of the invention may also be derived, i.e., with modifications other than insertion, removal or substitution of amino acid residues. Preferably, the modifications are covalently bonded in nature and include, for example, chemical bonding to the polymer, other organic and inorganic moieties of the lipid. The derivatives of the present invention can be prepared to extend the cycle of -46-016662 1316961 (42), the half-life of the invention, or the ability to modify the multi-peptide target to target cells, tissues or organs. The salvage receptor binding domain in the entire Fc molecule can also be used as the Fc portion of the compounds of the invention, as in WO 96/32478, entitled "Altered Polypeptides with Increased Half-Life". The other members of the class referred to herein as Fc are WO 97/34631, entitled "Immunoglobulin-Like Domains with Increased Half-Lives". The two published PCT applications cited in this paragraph. Both are incorporated herein by reference.

The Fc fusion can be at the N- or C-terminus of TMP1 or TMP2, or at the N- and C-termini of TMP1 or TMP2. Similarly, Fc fusion can be at the N- or C-terminus of the Fc functional site. Preferred compounds of the invention include IgGl Fc fusion dimers which are linked or otherwise followed by a dimer or multimer of the TMPs disclosed herein. In such instances, each Fc functional site is linked to a TMP dimer or multimer, with or without a linking agent. A schematic example of such a compound is shown in Figure 2. Multiple vehicles can also be used; such as having several Fc at each end or one Fc at one end and one PEG group at the other or side chain. Exemplary peptide-vehicle compounds are set forth in Table 4 below. Π1 -47- 1316961 (43) Table 4 - Example peptide-vehicle compound amino acid sequence SEQ ID NO: HIREGPTLRQWLVALRMV-GGG(PEG)GGG-HIREGPTLRQWLVALRMV 103 Fc-TCEQGPTLRQWLLCRQGR-GGGKGGG-TCEQGPTLRQWLLCRqGR-Fc 104 Fc-QLGHGPTLRQWLSWYRGM- GPNG-ELRSGPTLKEWLVWRLAq 105 CSWGGPTLKQWLQCVRAK-Fc 1 1 SWGGPTLKQWLQCVRAK 106 Fc-GGGKGGG-AVPQGPTLKQWLLWRRCA 107 PEG-CSSGGPTLREWLQCRRMQ I 1 CSSGGPTLREWLQCRRMQ 108 Fc-GGGGG-YCDEGPTLKQWLVCLGLQ-GGGGG-YCDEGPTLKQWLVCLGLQ 109 CSWGGPTLKQWLQCVRAK-GGGAGGG-CSWGGPTLKQWLQCVRAK-GGGAGGG- CSWGGPTLKQWLQCVRAK-GGGAGGG-Fc 110 VGIEGPTLRQWLAQRLNP- GGGCGGG-VGIEGPTLRQWLAQRLNP-PEG 111 Fc-ELRSGPTLKEWLVWRLAq-GGGG-ELRSGPTLKEWLVWRLAQ 112 Fc-ALRDGPTLKQWLEYRRQA-GGGKGGG-ALRDGPTLKQWLEYRRQA-Fc 113 Further, Table 5 lists preferred embodiments of the present invention. Table 5 - Specific examples of preferred specific amino acid sequence .SEQ ID NO: ALRDGPTLKQWLEYRRQA-ALRDGPTLKQWLEYRRQA 114 EALLGPTLREWLAWRRAQ-EALLGPTLREWLAWRRAQ 115 AVPQGPTLKQWLLWRRCA-AVPQGPTLKQWLLWRRCA 116 YCDEGPTLKQWLVCLGLQ-YCDEGPTLKQWLVCLGLQ 117 CSSGGPTLREWLQCRRMQ-CSSGGPTLREWLQCRRMQ 118 CSWGGPTLKQWLQCVRAK-CSWGGPTLKQWLQCVRAK 119 ALRDGPTLKQWLEYRRQA-GGGGG-ALRDGPTLKQWLEYRRQA 120 EALLGPTLREWLAWRRAQ- GGGGG-EALLGPTLREWLAWRRAQ 121 AVPQGPTLKQWLLWRRCA-GGGGG-AVPQGPTLKQWLLWRRCA 122 YCDEGPTLKQWLVCLGLQ-GGGGG-YCDEGPTLKQWLVCLGLQ 123 CSSGGPTLREWLQCRRMQ-GGGGG-CSSGGPTLREWLQCRRMQ 124 CSWGGPTLKQWLQCVRAK-GGGGG-CSWGGPTLKQWLQCVRAK 125 Fc-GGGGG-ALRDGPTLKQWLEYRRQA 126 Fc-GGGGG-EALLGPTLREWLAWRRAQ 127 Fc-GGGGG-AVPQGPTLKQWLLWRRCA 128 Fc-GGGGG- Fc-GGGGG-CSSGGPTLREWLQCRRM130 Fc-GGGGG-CSWGGPTLKQWLQCVRAK 131 48·0163^4 说明明说明 1316961 AVPQGPTLKQWLLWRRCA-GGGGG-A VPQGPTLKQWLLWRRCA 134 Fc-GGGGG-YCDEGPTLKQWLVCLGLQ-GGGGG-YCDEGPTLKQWLVCLGLQ 135 Fc-GGGGG-CSSGGPTLREWLQCRRMQ-GGGGG-CSSGGPTLREWLQCRRMQ 136 Fc-GGGGG-CSWGGPTLKQWLQCVRAK-GGGGG-CSWGGPTLKQWLQCVRAK 137 ALRDGPTLKQWLEYRRQA-GGGGG-ALRDGPTLKQWLEYRRQA-GGGGG-Fc 138 EALLGPTLREWLAWRRAQ-GGGGG-EALLGPTLREWLAWRRAQ-GGGGG- Fc 139 AVPQGPTLKQWLLWRRCA-GGGGG-AVPQGPTLKQWLLWRRCA-GGGGG-Fc 140 YCDEGPTLKQWLVCLGLQ-GGGGG-YCDEGPTLKQWLVCLGLQ-GGGGG-Fc 141 CSSGGPTLREWLQCRRMQ-GGGGG-CSSGGPTLREWLQCRRMQ-GGGGG-Fc 142 CSWGGPTLKQWLQCVRAK-GGGGG-CSWGGPTLKQWLQCVRAK-GGGGG-Fc 143 ALRDGPTLKQWLEYRRQA-GGGGG-Fc 144 EALLGPTLREWLAWRRAQ- GGGGG-Fc 145 AVPQGPTLKQWLLWRRCA-GGGGG-Fc 146 YCDEGPTLKQWLVCLGLQ-GGGGG-Fc 14 7 CSSGGPTLREWLQCRRMQ-GGGGG-Fc 148 CSWGGPTLKQWLQCVRAK-GGGGG-Fc 149 III. Method of Manufacture The compound of the present invention can be produced by various methods. Since many such compounds are peptides, or include peptides, the method of synthesizing peptides has a particular relevance herein. Solid phase synthesis techniques can be used. Suitable techniques are well known in the art, including those described in the following documents: Merrifield, in Chem. Polypeptides, pp. 335-61 (Katsoyannis and Panayotis eds. 1973); Merrifield, J. Am. Chem. Soc. 85: 2149 (1963); Davis et al.,

Biochem. Inti. 10: 394-414 (1985); Stewart and Young, Solid Phase Peptide Synthesis (1969); U.S. Patent No. 3,941,763; Finn et al., The Proteins, 3rd ed., vol. 2, pp. 105 -253 (1976); and Erickson et al., The Proteins, 3rd ed., vol. 2, pp. 257-527 (1976). Solid phase synthesis is a preferred technique for making individual peptides because it is the most cost effective method for making small peptides. The peptide can also be produced using recombinant DNA techniques in a transgenic host cell. To carry out this, a recombinant DNA molecule encoding the peptide is prepared. Methods of preparing such DNA and/or RNA molecules are well known in the art. For example, the sequence encoding the peptide can be cleaved from the DNA using an appropriate restriction enzyme -49-016665 1316961 (45). The formation of related sequences can be made using polymerase chain reaction (PCR) and adding useful restriction sites for subsequent selection. Alternatively, DNA/RNA molecules can be synthesized using chemical synthesis techniques, such as the phosphoramidite process. Combinations of these or other techniques can also be used. The invention also encompasses a vector encoding a peptide in a suitable host. The vector comprises a DNA molecule encoding the peptide which is manipulated to bind to an appropriate expression control sequence. The method of performing this operative linkage is well known before or after the DNA molecule encoding the peptide is inserted into the vector. Expression control sequences include promoters, activators, enhancers, ribosome binding sites, initiation-signals, stop signals, cap signals, polyadenylation signals, and other signals associated with transcriptional or translational control. The resulting vector comprising a DNA molecule encoding a peptide is used to transform a suitable host. This transformation can be carried out using methods well known in the art. Any of a large number of existing and well-known host cells can be used to practice the invention. The choice of a particular host will depend on the various factors identified by the art. These factors include, for example, compatibility with the selected performance vector, toxicity of the peptide encoded by the DNA molecule to the host cell, rate of transformation, ease of peptide recovery, performance characteristics, biosafety, and cost. To understand that not all hosts have the same effect on the performance of a particular DNA sequence, a balance of these factors must be achieved. Under these general guidelines, available microbial hosts include bacteria (such as E. coli), yeasts (such as Saccharomyces and Pichia pastoris), and other cultured fungi, insects, and plants. • Mammalian (including human) cells, or other hosts known in the art. The transformed host line is cultured under conventional fermentation conditions to exhibit a desirable peptide. Such fermentation conditions are well known in the art. Then spontaneous 016666 -50- 1316961 (46) «巍明续:! The peptide is purified from the fermentation medium or from the host cell expressing the peptide. Purification methods are also well known in the art. The compound containing the derived peptide, or the compound containing the non-peptidyl group, can be synthesized by well-known organic chemical techniques. For example, solid phase synthesis techniques can be used. Suitable techniques are well known in the art, including those described in Merrifield (1973), Chem. Polypeptides, ρρ. 335-61 (Katsoyannis and Panayotis eds.); Merrifield (1963), J. Am. Chem. Soc. 85:2149; Davis et al. (1985), Biochem. Inti. 10:394-414; Stewart and Young (1969), Solid Phase Peptide Synthesis; US Patent 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: 25 7-527. Solid phase synthesis is a preferred technique for making individual peptides because it is the most cost effective method for making small peptides. IV. Uses of Compounds The compounds of the invention have the ability to bind to and/or activate mpl receptors, and/or stimulate the production of platelets (in vivo and in vitro) ("platelet production activity'') and production of platelet precursors ("megakaryocyte production activity"). To measure the activity of these compounds, standard assays can be used, as described in WO 95/26746 entitled "Compositions and Methods for Stimulating Megakaryocyte Growth and Differentiation". In vivo assays are further described in the Examples section of this document. The conditions treated by the methods and compounds of the invention generally relate to existing megakaryocyte/platelet deficiency or megakaryocyte/platelet deficiency that may or may be expected in the future (e.g., due to planned surgery or platelet donation). Such conditions can be the result of a lack of active mpl ligand (temporary or permanent) in vivo. The lack of platelets is known as thrombocytopenia. Therefore, the methods and compounds of the invention can be used to prevent or treat thrombocytopenia in patients with sputum, sputum, sputum, and sputum. . The World Health Organization has classified the extent of thrombocytopenia according to the number of platelets A, , and platelets (Miller, et, 'a1, Cancer 47:210-211 (19 8 1)). For example, an individual who does not show a reduction in blood chicks (grade) usually has at least 100,000 platelets/cubic parachute 1; (level 1) shows a platelet level of sputum recognition, sputum depletion/inflammation. By wearing;, the method of 丨, W from 79,00 〇 to 99,000 / cubic milligrams of moderate to moderate thrombocytopenia (level 2) shows ▲ 50,000 to 74,000 / cubic millimeter, the severity of severe blood is between 25, 〇 〇〇 to 49,000Y cubic millimeters. Opposite, degenerative features characterized by thrombocytopenia characterized by a small threat of blood opening or a weak cubic millimeter. h ^ cycle concentration less than 25,0_ thrombocytopenia (platelet deficiency) also chemotherapy and other treatments using various drugs, for a variety of reasons, including blood loss and other specific diseases ^,,, surgical surgery, accidental According to the present invention, it can treat primary or immune platelets, sputum, and force. Aplastic anemia; \ syndrome (iTp,, - primary thrombocytopenia purple Insane; &> includes detective thrombocytopenic purpura associated with breast cancer; private ITP and HIV-related blood spleen, systemic lupus erythematosus; tumor metastasis including neoskeletal reduction;兄 狼 狼 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒 雒'Acute and severe hemorrhagic nocturnal hematuria after treatment with thrombocytopenia (Abciximab); C7E3 Fab plate reduction, including maternal alloimmunization J plate reduction; allogeneic immune corpuscles and thrombosis Platelet thrombocytopenia; anti-phospholipid immune platelet hypothyroidism; & ^ 01666$ '52. Description of the invention continuation page 1336961 (.) Symptoms, drug-induced thrombocytopenia, including carboplatin induced Thrombocytopenia, heparin-induced thrombocytopenia; fetal thrombocytopenia; any stage of thrombocytopenia; Hughes syndrome; lupus-like thrombocytopenia; incidental and/or massive blood loss; myeloid hyperplasia; Human thrombocytopenia; thrombotic thrombocytopenic purpura, including thrombotic microangiopathy in patients with cancer who develop thrombotic thrombocytopenic purpura/hemolytic urinary toxicity; autoimmune hemolytic anemia Recessive jejunal perforation; simple red blood cell hypoplasia; autoimmune thrombocytopenia; nephropathy epidemic; acute renal failure associated with rifampicin; Paris-Trousseau thrombocytopenia; neonatal allotype Thrombocytopenia; paroxysmal nocturnal hematuria; blood changes in gastric cancer; early childhood hemolytic uremic syndrome Blood symptoms associated with viral infections, including hepatitis A virus and CMV-related thrombocytopenia. In addition, some treatments for AIDS can also cause thrombocytopenia (such as AZT). Some wound healing disorders can also be derived from platelets. Benefit from an increase in the number. For the expected platelet deficiency, such as due to future surgery, the compounds of the invention can be administered from a few days to several hours before the need for platelets. For acute conditions, such as accidental and large For loss of blood, the compounds of the invention can be administered with blood or purified platelets.The compounds of the invention can also be used to stimulate certain cell types other than megakaryocytes, as long as such cells are found to exhibit mpl receptors. It is also within the scope of the present invention to correlate with such cells expressing mpl receptors and to respond to stimulation of mpl ligands. Oima -53- 1316961 (49) Description of the Invention - The compound of the present invention can be used for any kind of disease requiring the production of platelets or platelet precursor cells, or in which stimulation of the mpl receptor is required. Thus, for example, the compounds of the invention may be used to treat any condition in which a mammal requires platelets, megakaryocytes, and the like. Such conditions are described in detail in the following example sources: WO 95/26746; WO 95/21919; W0 95/18858; W0 95/21920, each of which is incorporated herein by reference. The compounds of the invention may also be used to maintain the viability or lifespan of platelets and/or megakaryocytes and associated cells. Accordingly, it is useful to include an effective amount of one or more such compounds in a composition comprising such cells. "Mammal" means any mammal, including humans, livestock, including dogs and depictions; exotic and/or zoo animals including monkeys; laboratory animals including mice, rats, and guinea pigs; farm animals including horses, cattle, Sheep, goats and pigs; and the like. Preferred mammals are humans. V. Pharmaceutical Compositions The invention also provides pharmaceutical compositions and methods of using the pharmaceutical compositions of the compounds of the invention. Such pharmaceutical compositions can be administered for injection. Or oral, nasal spray, subcutaneous injection or other forms of administration, including, for example, by intravenous injection, intradermal injection, intramuscular injection, intramammary injection, intrapulmonary injection (eg, aerosol) or subcutaneous injection (including storage) For long-term release; for sublingual, anal, vaginal, or surgical transplantation, such as transplantation under the spleen sac, brain or cornea. Treatment can consist of a single agent or multiple agents over a period of time. Generally speaking, The pharmaceutical compositions encompassed by the present invention comprise an effective amount of a compound of the invention and a pharmaceutically acceptable diluent, preservative, solubilizer, emulsifier, Agent and / or carrier. Such compositions include various -54-0166 ^ 0 (50) 1316961

Buffer content (eg Tds-HC, acetate, sputum phosphate), pH and ionic diluents, additives such as detergents and solubilizers (eg Tween80 poly 2^_^^ ("Blood blood Acidic acid, hard acid, chlorophyll, anti-fouling agent (such as Thimersol, benzyl alcohol) and volume expansion materials (such as lactose, mannitol); granular preparations of polymer compounds such as polylactic acid, polyglycolic acid, etc. Adding substances to the inside, or to the vesicles. It can also be: osmotic acid, which can have the effect of promoting residence time in the circulation. Pharmaceutical composition: including other pharmaceutically acceptable liquids, semi-solids as appropriate Or a solid second release agent's as a pharmaceutical vehicle, excipient or medium, including but not limited to, polyoxyethylene sorbitan monolaurate, magnesium stearate, thiobenzoic acid methyl And propyl esters, starch, sucrose, dextrose, gum arabic, calcium phosphate, mineral oil, cocoa butter and cocoa butter. Such compositions can affect the physical state, stability and in vivo of the proteins and derivatives of the invention. Release rate, and body clearance rate. See, for example, Remingt On,s pharmaceutieal

Sciences, 18th Ed. (1990, Mack Publishing Co., Easton, PA 18042) 14 3 5 - 1712 pages, which is incorporated herein by reference. The composition may be prepared in liquid form, or in dry powder form, such as in the form of a bead. Transplantability sustained release formulations are also contemplated, such as transdermal formulations. It is contemplated for use in the present invention as an oral solid dosage form, which is generally described in the following documents: Remington's Pharmaceutical Sciences, 18th Ed. 1990 (Mack Publishing Co., Easton, PA 18042) Chapter 89, by way of citation Incorporated herein. Solid dosage forms include lozenges, capsules, pills, tablets or granules, cachets or granules. The vesicles or proteinaceous vesicles can also be used to formulate compositions of the invention (e.g., protein-like microspheres, available in U.S. Patent No. 01,667, filed on Jan. No. No. 4, 925, 673). Liposomes can be used, and the vesicles can be derivatized with various polymers (e.g., U.S. Patent No. 5,013,556). A description of possible solid dosage forms for treatment is given by Marshall, K., Modern Pharmaceutics, Edited by G. S. Banker and C. T. Rhodes Chapter 10, 1979, which is incorporated herein by reference. In general, the formulations comprise a compound of the invention, together with an inert ingredient which provides protection in the stomach environment and which releases the biologically active substance in the intestine. Oral dosage forms of the above compounds of the invention may also be specifically contemplated. If necessary, the compound can be chemically modified to render oral delivery effective. In general, a chemical modification that can be considered is the incorporation of at least a portion of the body to the compound molecule itself, wherein the portion can contribute to (a) inhibition of proteolysis; (b) uptake into the bloodstream from the stomach or intestine. It is also desirable to increase the overall stability of the compound and to extend its circulation time in the body. Examples of such partial bodies include: polyethylene glycol, copolymers of ethylene glycol and propylene glycol, carboxymethyl cellulose, dextran, polyethylene glycol, polyethyl hydrazine p than terpene ketone, and polyglycine ( 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 useful polymers are poly-1,3-dioxolane and poly-1;3,6-trioxolane. For medical use, as described above, it is a polyethylene glycol moiety. For oral delivery dosages, it is also possible to use modified aliphatic amine salts such as N-(8-[2-hydroxybenzylidenyl]amino)octanoate (SNAC),

As a carrier for increasing the absorption rate of the therapeutic compound of the present invention. The clinical effectiveness of the use of SNAC 016672 -56 - 1316961 (52) description of the continuation of the heparin formulation has been confirmed by Emisphere Technologies in the second phase of the trial. See U.S. Patent 5,792,451 ' " Oral drug delivery composition and methods" 0 The therapeutic agent can be included in the formulation as fine multiparticulates in the form of granules or pellets having a particle size of about 1 mm. The formulation of the substance to be used for capsule administration may also be a powder, a lightly compressible suppository or even a lozenge. The therapeutic agent can be prepared by compression. Both colorants and flavor enhancers can be included. For example, the protein (or derivative) can be formulated (e.g., trans-lipid or microsphere capsules) and then further included in an edible product, such as a frozen beverage containing a colorant and a flavor enhancer. It can be used as an inert substance to dilute or increase the volume of the therapeutic agent. These diluents may include: vinegars 'especially mannitol, lactose, anhydrous lactose, cellulose, inexpensive sugars' modified dextran and starch. Certain inorganic salts can also be used as fillers' including sodium trisodium sulphate, magnesium carbonate and sodium chloride. Some of the available dilutions are Fast Flo, Emdex, STA-Rx 1500, Emcompress and Avicell. The Φ disintegrating agent can be formulated into a solid dosage form in the formulation of the therapeutic agent. Substances useful as disintegrants include, but are not limited to, starch, including commercially available disintegrants based on decidation, 'Expl〇tab. Sodium starch glycolate, Amberlite, sodium methicillin, hyperamylopectin, sodium alginate, gelatin, orange peel, acid carboxymethylcellulose, natural sponge and bentonite can be used. Another form of disintegrant is an insoluble cation exchange resin. Powdered gums can be used as disintegrants and binders, and include powdered gums such as agar, paulownia gum or tragacanth. Alginic acid and its sodium salt can also act as disintegrants. -57- 016673 Description of the Invention Continued Page 1316961 (53) Adhesives may be used to bond the therapeutic agents together to form a hard lozenge, and include materials derived from natural products such as gum arabic, gums, temples, and gelatin. Others include mercapto cellulose (MC), ethyl cellulose (EC), and carboxymethyl cellulose (CMC). Both polyvinylpyrrolidone (PVP) and hydroxypropylmethylcellulose (HPMC) can be used in an ethanol solution to granulate a therapeutic agent. Anti-friction agents may be included in the formulation of the therapeutic agent to avoid sticking during formulation. Lubricants can be used as a barrier between the therapeutic agent and the mold wall. These can include, but are not limited to, stearic acid, including magnesium stearate and calcium stearate, polytetrafluoroethylene (PTFE), liquid paraffin, vegetable oils. And wax. Soluble lubricants such as sodium lauryl sulfate, magnesium lauryl sulfate, polyethylene glycols of various molecular weights, Carbowax 4000 and 6000 〇 can also be added to improve the fluidity of the drug during formulation and aid in the compression process. A regurgitant for reconstitution. Glidants include starch, talc, high-temperature Shishi and hydrated seconds. To aid in the dissolution of the therapeutic agent in an aqueous environment, a surfactant can be added as a wetting agent. Surfactants include anionic detergents such as sodium lauryl sulfate, sodium octyl sulfosuccinate and sodium dioctyl sulfonate. A cationic detergent can be used and can include decyl ammonium chloride or acetaminophen. A list of potential nonionic detergents that can be included as a surfactant in the formulation are lauryl polyethylene glycol 400, polyoxyl 40 vinegar, polyoxyethylidene castor oil 10, 50 and 60, glycerol alone. Stearate, polysorbate 40, 60, 65 and 80, sucrose fatty acid ester, methyl cellulose and carboxymethyl cellulose. These surfactants may be present in the formulation of the protein or derivative, either alone or in a mixture of different ratios. mMU -58- 1316961

(54) L5 Description L. Additives which may increase the uptake of the compound are, for example, lipoic acid, oleic acid, linoleic acid and linolenic acid. Controlling the release formulation is desirable. The drug can be incorporated into an inert matrix which can be released by diffusion or leaching mechanisms such as gums. Slowly metamorphic matrices can also be added to the formulation, such as sputum acid | S, sucrose. Another controlled release form of this therapeutic agent is through a method based on the 0r〇s treatment system (Alza Corp.), which also encapsulates the drug into the semipermeable membrane to allow water to enter and penetrate through a single small penetration. The opening drains the drug. Certain enteric coatings also have an extended release effect. Other coatings can be used in the formulation. These include various sugars which can be applied to the coated pan. The therapeutic agent can also be given in the form of a film-coated lozenge and the materials used in this example are classified into two classes. The first category is non-intestinal rolling material' and includes methyl cellulose, ethyl cellulose, hydroxyethyl cellulose, methyl ethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose 'Several methyl cellulose, providone and polyethylene glycol. The second type of enteric material is a commonly used phthalate. A mixture of materials can be used to provide an optimal film coating. Membrane coating may be in a disc coater or in a fluidized bed or through a compression coated form ##° The invention also encompasses pulmonary delivery of the protein of the invention (or a derivative thereof). The protein (or derivative) is delivered to the lungs of the mammal upon inhalation' and passes through the lung epithelial layer into the bloodstream. (Other reports on this include Adjei et aL, Pharmaceutical Research 7: 565-569 (1990); Adjei et al. 5

International Journal of Pharmaceutics 63: 135- 144 (1990) (leuprolide acetate); Braquet et al., Journal of Cardiovascular • 59-

1316961 (55)

Pharmacology 13 (suppl. 5); s. 143-146 (1989) (endothelin-1); Hubbard et al., Annals of Internal Medicine 3:206-212 (1989) (1-anti-pancreas Protease (i_antitrypsin)); Smith et al., J. CHn. Invest 84: 1145-1146 (1989) (1-proteinase); Oswein et al., "Aerosolization of Proteins ", Proceedings Of

Symposium on Respiratory Drug Delivery II, Keystone, Colorado,

March, 1990 (Recombinant Human Growth Hormone); Debs et al., The Journal of Immunology 140: 3482-3488 (1988) (Interferon and Tumor Necrosis Factor) and Platz et al., US Patent No. 5,284,656 (granular cells) Community stimulating factor). In the practice of the present invention, the use of mechanical devices designed to deliver therapeutic products to the lungs, including but not limited to nebulizers, graduated drug inhalers, and powder inhalers, is encompassed by all of these materials. Some of the examples of commercially available equipment for use in the practice of the present invention are Ultravent sprayers manufactured by Mallinckrodt, Inc., St. Louis, Missouri; Acorn II sprayers, by Marquest Medical Products, Englewood, Made in Colorado; Ventolin with graduated drug inhaler, manufactured by Glaxo Inc., Research Triangle Park, North Carolina; and Spinhaler powder inhaler, manufactured by Fisons Corp., Bedford, Massachusetts. 0 All of these devices require the use of compounds suitable for partitioning the present invention. Formulation. Typically each formulation will be specific to the type of device used and may include the use of a suitable aerosol matrix in addition to the diluent ' adjuvant and/or carrier available in the treatment. • 60· 016676 1316961 (56) Description of the Invention Continued Pages The compounds of the present invention are most advantageously prepared in a granular form having an average particle size of less than 10 microns, preferably 0.5 to 5 microns, for most efficient delivery to the lung distal end. . Carriers include cool materials such as trehalose, mannitol, xylitol, inexpensive sugar, lactose and sorbitol. Other ingredients that can be used for formulation include DPPC, DOPE, DSPC, and DOPC. Natural or synthetic surfactants can be used. Polyethylene glycol can be used (even in addition to its use in derivatized proteins or the like). Glucan can be used, such as cyclosaccharide. Bile salts and other related enhancers can be used. Cellulose and cellulose derivatives can be used. An amino acid can be used, for example, in a buffer to prepare a mist. In addition, it is also contemplated to use vesicles, microcapsules or microspheres, inclusion complexes, or other types of carriers. Formulations suitable for use with a spray or ultrasonic type nebulizer typically comprise a compound of the invention dissolved in water at a concentration of from about 0.1 to 25 mg of bioactive protein per ml of solution. Formulations may also include a buffer and a simple sugar (e.g., for protein stabilization and osmotic pressure adjustment). The nebulizer formulation may also contain a surfactant to reduce or avoid surface induced aggregation of the protein caused by the aerosol formation of the aerosol upon solution. Formulations for use with graduated medicinal inhaler devices typically comprise a finely divided powder comprising a compound of the invention which is suspended in an aerosol matrix by means of a surfactant. The aerosol matrix can be any conventional material for this purpose, such as chlorofluorocarbons, hydrochlorofluorocarbons, hydrofluorocarbons, or hydrocarbons, including trichlorofluoromethane, dichlorodifluorodecane, dichloro Tetrafluoroethylene-61 - 016677 1316961 (57) Description of the invention cont. j Alcohol and 1,1,1,2-tetrafluoroethane, or a combination thereof. Suitable surfactants include sorbitan trioleate and soy lecithin. Oleic acid can also be used as a surfactant. Formulations dispensed from a powder inhaler device include finely divided dry powders containing a compound of the invention, and may also include bulking agents such as lactose, sorbitol, sucrose, mannitol, trehalose or xylitol in amounts effective to aid The powder from the device is dispersed, such as from 50 to 90% by weight of the formulation. Nasal delivery of the compounds of the invention is also contemplated. Nasal delivery allows the protein to enter the bloodstream directly after nasal administration of the therapeutic product without the need to deposit the product in the lungs. Nasal delivery formulations include those containing dextran or cyclamate. Delivery using carriers that pass through other mucosa is also covered. Dosage The method of pharmacy included in the method of treating the above-mentioned diseases is determined by the doctor who visits, and will consider various factors that alter the action of the drug, such as the age, physical condition, weight, sex and diet of the patient, the severity of the infection, and the application. The time of the drug and other clinical factors. The administration of the compounds of the invention may be preceded by the use of a pill followed by continuous infusion to maintain a therapeutic circulating level of the drug. In another embodiment, the compounds of the invention may be administered in a single dose.谙 This technique can be easily optimized for effective medical procedures and clinical conditions of individual patients to optimize the effective dose and regimen. The frequency of administration depends on the pharmacokinetic parameters of the agent and the route of administration. The best pharmaceutical formulation can be determined by the skilled artisan according to the route of administration and the desired dosage. See, for example, Remingtoi^s Pharmaceutical Sciences, 18th Ed. (1990, Mack Publishing Co., Easton, PA 18042) - 62-016678, pp. 1316961 (58) pages 1435-1712, the disclosure of which is incorporated herein by reference. . Such formulations can affect the physical state, stability, rate of in vivo release, and rate of in vivo clearance of the agent being administered. Depending on the route of administration, the appropriate dose can be calculated based on body weight, body surface area or volume of the organ. Further precision in the calculations required to determine the appropriate agent for treatment with each of the above formulations may be routinely accomplished by the skilled artisan without undue experimentation, particularly in accordance with the dosage information and assays disclosed herein, and in Pharmacokinetic data observed in the above human clinical trials. The appropriate dose can be determined by the existing assay for determining the blood level dose together with the appropriate dose-response data. The final pharmacy will be determined by the attending physician, considering various factors that alter the action of the drug, such as the specific activity of the drug, the severity of the injury and the patient's responsiveness, the patient's age, physical condition, weight, sex and diet, any The severity of the infection, the time of administration and other clinical factors. As the study progresses, further information is available on appropriate dose levels and duration of treatment for various diseases and conditions. Therapeutic methods, compositions and compounds of the invention may also be used, either alone or in combination with other interleukins, soluble mpl receptors, hematopoietic factors, interleukins, growth factors or antibodies for the treatment of other symptoms and thrombocytopenia Characteristics of the disease state. It is expected that the compounds of the invention will prove useful in combination with common hematopoietic stimulators such as IL-3 or GM-CSF for the treatment of certain forms of platelet reduction. Other megakaryocyte stimulating factors such as meg-CSF, stem cell factor (SCF), leukemia inhibitory factor (LIF), oncostatin M (OSM), or other molecules with megakaryocyte stimulating activity may also be associated with mpl Use together. Other 016679 -63 - 1316961 (59) Description of the Invention for continued use of such interleukins or hematopoietic factors include IL-1 alpha, IL-1 /3, IL-2, IL-3, IL- 4, IL-5, IL-6, IL-U, community stimulating factor - l (CSF-l), M-CSF, SCF, GM-CSF, granulocyte community stimulating factor (G-CSF), EPO, interferon -a (IFN-α), consensus interferon, IFN-dots, IFN-r, IL-7, IL-8, IL-9, IL-10, IL-12, IL-13, IL-14, IL- 15, IL-16, IL-17, IL-18, thrombopoietin (TPO), angiopoietin such as Ang-1, Ang-2, Ang-4, Ang-Y, human angiopoietin peptide, Vascular endothelial growth factor (VEGF), angiogenin, bone type protein-1, bone form egg-white matter-2, bone form protein-3, bone form protein-4, bone Forming protein-5, bone form protein-6, bone form protein-7, bone form protein-8, bone form protein-9, bone form protein-10, bone type Protein-11, bone morphogenetic protein -12, bone type protein-13, bone form protein-14, bone form protein-15, bone form protein receptor IA, bone form protein receptor IB, brain-derived neurotrophic Factor, cilia trophic factor, cilia trophic factor receptor, interleukin-induced neutrophil chemokine 1, interleukin 谤 neutrophil chemokine 2, endothelial cell growth factor, endothelin 1, dermal growth Factor, epithelial-derived neutrophil jk ball attractant, fibroblast growth factor 4, fibroblast growth factor 5, fibroblast growth factor 6, fibroblast growth factor 7, fibroblast growth factor 8, fibroblast Cell growth factor 8b, fibroblast growth factor 8 c, fibroblast growth factor 9, fibroblast growth factor 10, fibroblast growth factor acidity, fibroblast growth factor test, God-64-016680 1316961 ( 60) Description of the invention Continued: Glial cell line-derived neurotrophin receptor 1, glial cell line-derived neurotrophin receptor 2, growth Proteins, growth-related proteins, growth-related proteins, growth-related proteins, heparin-linked dermal growth factor, hepatocyte growth factor, hepatocyte growth factor receptor, insulin-like growth factor I, insulin-like growth factor Body, insulin-like growth factor II, insulin-like growth factor binding protein, keratinocyte growth factor, leukemia inhibitory factor, leukemia inhibitory factor incineration, nerve growth factor, nerve growth factor receptor, neurotrophin-3, neurotrophin 4 'Placental growth factor, placental growth factor 2, platelet-derived endothelial cell growth factor, platelet-derived growth factor, platelet-derived growth factor Α bond, platelet-derived growth factor ΑΑ, platelet-derived growth factor AB, platelet-derived growth Factor B bond, platelet-derived growth factor BB, platelet-derived growth factor receptor, platelet-derived growth factor receptor, pre-B cell growth stimulating factor, stem cell factor receptor, TNF, including TNF0, TNF1, TNF2, transformation Growth factor, transforming growth factor, transforming growth Long factor 1, transforming growth factor 1.2, transforming growth factor 2, transforming growth factor 3, transforming growth factor 5, recessive transforming growth factor 1, transforming growth factor binding protein I, transforming growth factor binding Protein II, transforming growth factor binding protein III, tumor necrosis factor receptor type I, tumor necrosis factor receptor type II, urokinase plasminogen activator receptor, vascular endothelial growth factor, and inlay Synthetic proteins and their biological or immunologically active fragments. It is also useful to simultaneously or sequentially administer an effective amount of a soluble mammalian mpl receptor, which has the effect of causing megakaryocyte segmentation to form platelets when the megakaryocytes reach a mature state. Therefore, give this hair -65- 1316961

(61) Ming compounds (to increase the number of mature megakaryocytes) followed by the use of soluble mpl receptors (to de-ligand activity and to produce platelets from mature megakaryocytes) are expected to be a particularly effective way to stimulate platelet production. The above dosages can be adjusted to compensate for such additional ingredients in the therapeutic composition. The progress of the treated patient can be monitored by conventional methods. In the case where the compound of the present invention is to be added to a composition of platelets and/or megakaryocytes and related cells, the amount to be included is usually determined experimentally by techniques and assays known in the art. An example of this amount ranges from 0.1 microgram to 1 milligram of the compound of the invention per 1 〇 6 cells. It will be appreciated that the application of the present invention to a particular problem or situation is intended to be within the skill of the subject skilled in the art. Examples of the products of the invention and representative methods for their isolation, use and manufacture are as follows. EXAMPLES The following describes the exemplary manufacturing and identification methods of the compounds disclosed herein. Example 1 1. Composition of the secondary peptide library Α·Preparation of the competent colorectal-like cells: The isolated colonic bacterium (TG1 strain) Amersham Pharmacia Biotech, Piscataway, New Jersey) Cultures were prepared at 10 °C in 10 ml of 2xYT medium (1.6% Bacto Tryptone, 1% yeast extract, 85.5 mM NaCl). Use one milliliter of this overnight culture for inoculation! 2 χγτ medium containing 0.4% glucose and 1 mM MgCl2, and the one liter 016682 • 66 - (62) 1316961 nutrient grows in the spreader at 37C to 〇〇6 ==〇8 . The culture was chilled on ice for 15 minutes and detached from the knife at 4 rpm (Beckman JA_1 〇 rotor) for 2 minutes. Resuspend the bacterial pellets in 5 ml of ice 々10/. Glycerol was mixed in and the resulting mixture was at (10) rpm at 4. (: From the 'U knife away from the 2 knives 4 liters. The bacteria pellets are resuspended in 500 ml of ice-cold 10% gan' from the solution, and the resulting mixture is again centrifuged with 4 〇〇〇卬 claws at 41 #20刀I. The cell pellet was then resuspended in 25 ml of ice-cold 〇% glycerol solution. The concentrated bacterial sample was transferred to an ice-cold 5 〇 ml conical tube, and centrifuged on the table. Centrifuge in a Beckman CS-6R at 3 500 rpm for 15 minutes at 4 C. Resuspend the pellet in a small volume of ice-cold glycerol solution and add 1 〇〇 or 3 〇〇 microliters. The bacterial stock solution was immediately frozen in an ethanol/dry ice bath and stored in a freezer at -8 ° C. B. Improvement of the pCES1 vector

PCR was carried out using Extend Long Template PCR Systems (Roche Diagnostics Corp., Indianapolis, Indiana) with 1 μg of pCES1 vector (TargetQuest Inc.) as a template. The volume of the PCR mixture was 100 μl, including 1 X PCR buffer, two primers of 200 nM each 5'-CAAACGAATGGATCCTCATTAAAGCCAGA-3' ^ 5'-GGTGGTG CGGCCGCACTCGAGACTGTTGAAAGTTGTTTAGCA-3', 200 nM dNTP, 3 Units of Tag DNA Polymerase. The following procedure was run using the TRIO-Thermoblock (Biometra) PCR system: 5 minutes at 94 ° C; 30 cycles (30 seconds at 94 ° C, 30 seconds at 50 ° C, 45 seconds at 72 ° C); 72 ° C for 10 minutes; cool to 4 °C. The PCR product was run on a 1% agarose gel and purified using a QIAGEN Spin Column according to the manufacturer's instructions -67· 016683 1316961 (63) Description of the Invention Continued on: (QIAGEN Inc., Valencia, California). The second PCR reaction was carried out under the same PCR conditions as described on port i using 5 μl of the PCR product and 2 n of each of the 200 nM primers 5'-CAAACGAATGGATC CTCATTAAAGCCAGA-3' and 5'-AACAAAAAGTGCACAGGGTGGA GGTGGTGGTGCGGCCGCACT-31. The PCR product and the original pCES1 vector were separately in 1 〇〇 microliter of reaction solution containing 'xx NEB2 buffer, 60 units of ApaLI (New England Biolabs, Beverly, Massachusetts), 60 units of BamHI (New England) Biolabs), digestion at 37 ° C for 1 hour. The two digested DNAs were purified using QIAGEN Spin Column and ligated overnight in 40 μl of reaction mixture containing lx ligation buffer and 40 units of T4 DNA ligase (New England Biolabs). ). The vector was transfected into E. coli and incubated overnight at 37 °C. The single drop was chosen as the exit and the plastid was purified using the QIAGEN Spin Column. The correct insertion is determined by the DNA sequence. C. The rate of vector DNA,

One microgram of the modified pCES1 vector DNA (paragraph 1B) was transformed into 100 microliters of electricity competent TG1 E. coli (ία segment) using Gene Pulser II (BIO-RAD, Hercules, California), set to 2500 Volt (V), 25T, and 200 ohms. The transformed bacterial sample was then immediately transferred to 900 μl of SOC (2% pancreas, 0.5% yeast extract, 1 mM sodium chloride, 2.5 mM potassium chloride, 20 mM glucose, 1 In a test tube of mM mM magnesium sulfate, 10 mM cesium chloride), the culture was grown under shaking at 37 for 1 hour. The cells were then dispersed into 2XYTAG (2xYT, containing 100 micro-68-1316961 _ (64) invention storage kg / ml ampicillin and 2% glucose) agar plates, and incubated at 37 ° C night. One liter of 2XYTAG medium was inoculated overnight at 37 ° C using a single flora. The plastid vector DNA was purified using QIAGEN Plasmid Maxi Kit according to the manufacturer's instructions. β D. Digestion of vector DNA 50 μg of vector DNA (Phase 1C) in 400 μl of 2,200 units of ApaLI and 200 units containing lx NEB buffer In the Xhol reaction solution, it was digested overnight at 37 °C. The restriction digestion reaction solution was incubated at 37. (: All night and analyzed in a prefabricated 1% agarose gel (Embi Tec, San Diego, California). The linearized vector DNA was cleaved from the gel and used with QIAquick Gel Extraction Kit (QIAGEN Inc.) Extraction according to the manufacturer's instructions E. Preparation of the oligomeric nucleoside acid library Design of two oligonucleotide libraries (immobilized and additive-added). Synthesis of immobilized library oligonucleoside 5' -CACAGTGCACAGGGTNNKNNKNNKNNKGGTCCTACTCTGMR KSARTGGCTGNNKNNKNNKNNKNNKNNKCATTCTCTCGAGATC G-3', and the additive-incorporated oligo-nucleotide 5'-CACAGTGCAC-AGGGTNNKNNKNNKNNKggKcc-KacKctKNN KNNKtgKNNKNNKNNKNNKNNKNNKNNKCATTCTCTCGAGATC G-3H lowercase letters represent one of the indicated bases and one of the other three nucleotides a mixture of components). These oligonucleotides were used as templates in the polymerase chain reaction, respectively. The Expand High Fidelity PCR System kit (Roche Diagnos sties Corp.) was used in the PCR reaction. Each PCR reaction is in a volume of 100 μl, and 016685 -69 - Illustrated in the Continuation "ΙΊ 1316961 (65) oligonucleotide library containing 10 ηΜ, IX PCR buffer, each primer of each 300 ηΜ, -CACAGTGCACAGGGT-3' and 5'-TGATCTCGAGAGAATG -3', 200 nM dNTP, 2 mM calcium chloride and 5 units of Expand polymerase. A thermal cycler (GeneAmp PCR System 9700, Applied Biosystem) was used to perform the following procedure: 5 minutes at 94 ° C; 30 cycles (30 seconds at 94 ° C, 30 seconds at 55 ° C, 45 seconds at 72 ° C) ); 7 minutes at 72 ° C; cooled to 4 ° C. Use the QIAquick Nucleotide Removal Kit (QIAGEN Inc.) to remove free nucleotides according to the manufacturer's instructions. F. Digestion of the oligonucleotide library 5 micrograms of each PCR product (paragraph 1E) in 400 microliters of reaction solution containing lx NEB buffer, 2,200 units of ApaLI and 200 units of Xhol, at 37 Digested at °C overnight. The digested DNA was separated on a 3% agarose gel (Embi Tec). The target DNA bands obtained from each reaction solution were excised from the gel and extracted with a QIAquick Gel Extraction Kit. G. Ligation of the vector linearization vector (1D segment, 25 μg) and each digested PCR product (1F segment, 5 μg) in 400 μl of lx NEB ligation buffer and 80 units using an oligonucleotide library The reaction solution of T4 DNA ligase was ligated overnight at 16 °C. The ligation product was incubated at 65 °C for 20 minutes to inhibit activation of the DNA ligase, and then incubated with 8 units of Notl at 37 °C for 2 hours to minimize vector autologous ligation. The ligation product was then purified using standard phenol/chloroform extract (Molecular Cloning, Maniatis et al 3Td Edition) and resuspended in 30 microliters of water. H. Electrical Penetration -70- 01668 Liao 1316961 (66) Description of the Invention Continued _ Ten electrical penetration reactions were performed for each library. In each transformation, 3 microliters of ligation vector DNA (1G segment) and 300 microliters of TG1 cells (1A segment) were mixed in a 0.2-centimeter tube (BIO-RAD). The resulting mixture was pulsed with a Gene Pulser II set at 2500 volts, 25 uF and 200 ohms. The transformed bacterial samples obtained from the ten electroporation reactions were combined and transferred to a flask containing 27 ml of SOC and incubated at 37 ° C for 1 hour. The cells were then added to 170 ml of 2xYTAG and grown for 3 hours at 37 °C with shaking. The cells were centrifuged at 5000 rpm for 10 minutes at 4 °C. The pellet was then resuspended in 10 ml of 15% glycerol/2xYT and stored at -80 °C. This is the initial stock of the library. The titer showed that the library size was 1.0 X 1〇9 independent transform strain and 2.4 X 109 independent transform strain in the immobilized library and the additive library. 2. Amplification of the library A. Preparation of the secondary reservoir of the library The initial library cell stock (1H segment) was used to inoculate 1300 ml (for immobilization) and 2600 ml (for dosing) 2xYTAG medium. The starting OD6 〇 Q = 0.1. The culture was allowed to grow at 37 ° C for several hours until OD600 = 0.5. A 120 ml portion of the immobilized reservoir and a 240 ml portion of the pooled solution were taken and regenerated in a separate flask for 2 hours at 37 °C. These subcultures were centrifuged at 5000 rpm (Beckman JA-14 rotor) for 10 minutes at 4 ° C, and the pellets were resuspended in 10 ml (for each pool) of 15% glycerol/2xYT, and Store at -80 °C. B. Seedling induction M13K07 helper bacterium (Amersham Pharmacia Biotech) -71 - 016G87 1316961 vine (67) was added to the remaining bacterial culture (stage 2A) with OD6 〇 Q = 0.5 to a final concentration of 3 X 109 pfu/ml. The helper phage was infected with bacteria for 30 minutes at 37 ° C without shaking and slowly infected for 30 minutes. The infected cells were centrifuged at 5 ° rpm for 1 min at 4 °C. The cell pellet was resuspended in 1300 ml (immobilized pool) and 2600 ml (dosing pool) of 2xYTAK (2YT and 100 μg/ml ampicillin and 40 μg/ml kanamycin). At 37C, it was shaken overnight to produce phagemid. C. Phage harvesting The bacterial culture (Section 2B) was centrifuged at 5000 rpm for 10 minutes at 4 °C. The supernatant was transferred to a new bottle, 0.2 volume of 20% PEG/2.5 Μ sodium chloride was added, and incubated on ice for 1 hour to precipitate the phage. The precipitated phage was centrifuged at 8000 rpm at 4 ° C for 20 minutes and carefully resuspended in 1 mL of cold PBS. The phage plastid solution was separated by centrifugation at 8000 rpm for 10 minutes at 4 ° C, and 0.2 vol of 20% PEG / 2.5 Μ sodium chloride was added to precipitate the phage for further purification. The phage was centrifuged at 8000 rpm for 4 minutes at 4 ° C, and the phage was resuspended in 12 ml of cold PBS. 4 ml of a 60% glycerol solution was added to the phage plastid solution and stored at -80 °C. The titer of the phage plastid was determined by standard procedures (Molecular Cloning, Maniatis et al. 3rd Edition). 3. Selection of human MPL-binding phage A. Biotinylation of human MPL 1 mg of recombinant human MPL using EZ-LinkSulfo-NHS-LC-Biotinylation Kit (PIERCE, Rockford, Illinois) according to manufacturer's instructions -72- 016688 1316961 (68)

DESCRIPTION OF THE INVENTION The performance page J is biotinylated. B. MPL Stabilized biotinylated MPL on the magnetic sphere (3A segment) was immobilized on Dynabead M-280 Streptavidin (DYNAL, Lake Success, New York) at a concentration of 1 μg MPL/100 micro l from the manufacturer's pellets. After the beads were guided to one side of the tube with a magnet and the liquid was aspirated, the magnetic beads were washed twice with phosphate buffered saline (PBS) and resuspended in sulphate buffered saline. The biotinylated MPL protein was added to the washed magnetic beads at the same concentration as above and incubated at room temperature for 1 hour. BSA was then added to a 2% final concentration to block the MPL coated magnetic beads and incubated overnight at 4 °C. The resulting MPL coated magnetic beads were then washed twice with PBST (phosphate buffered saline and 〇.〇5 % Tween-20) and subjected to a selection step. C. Using MPL coated magnetic rosette for cleavage with 1 ml of 2% BSA in PBS blocked about 1 〇〇 of the library equivalent phage (2C segment 'lxlOncfu for immobilization library, 2.4xlOncfu for doping加库) 1 hour. A negative selection step was performed on the blocked phage sample, which was added to the empty beads (the same beads as the 3B segment but not coated with MPL)' and the mixture was rotated at room temperature Cultivate for 1 hour. The supernatant containing the sputum plastids was extracted by a magnet to a new tube containing MPL coated magnetic beads (3 B) and the mixture was incubated at room temperature for 1 hour. After the supernatant was removed, the phage-bound magnetic beads were washed 10 times with PBST and washed 1 time with PBS. The phages were then solubilized in 1 ml of 100 mM triethylamine solution (Sigma, St. Louis, Missouri) in a rotator for 10 minutes. The pH of the solution containing the sputum sputum is neutralized by adding 5·5 ml of 1 -73 - 016689 1316961 (69) Illustrated in the continuation of the elbow 1^-11 (:1 (?117.5). The plastids were infected with 5 ml of newly grown TG1 bacteria (0D6Q() for about 0.5) for 30 minutes without shaking at 37 °, and then slowly shaken for 30 minutes. All infected TG1 cells were plated on a large 2xYTAG plate. The plates were incubated and incubated overnight at 30 ° C. D. Induction and harvesting of sputum bacteria 10 ml of 2xYTAG medium was added to the plates (3C) to resuspend TG1 cells. All TG1 cells were collected. In a test tube, 250 μl of these cells were added to 25 ml of 2xYTAG and grown at 37 ° C until OD6()() = 0.5. The M13K07 helper phage was added to a final concentration of 3 X 109 Cfu/ml and incubate for 30 minutes without shaking at 37 ° C, then incubate for 30 minutes with gentle shaking. The cells were centrifuged at 5000 rpm for 10 min at 4 ° C and resuspended in 25 ml of 2x YTAK. The cells were shaken and grown overnight at 30 ° C. The phagosomes guided were harvested and purified as described in Section 2C. The second round selects the second round of selection as described in paragraphs 3B to 3C, with only the following differences. About 0.5 ml of the phage solution obtained from the 3D section is used as the feed phage Only 0.1 micrograms of biotinylated MPL (3A segment) was used to coat Dynabead M-280 Streptavidin. The p stem-host-magnetic beads were washed 16 times with PBST, where the final wash included at room temperature Incubate for 30 minutes in PBST. Wash the magnetic beads 10 times with PBS. 4. Analysis of the colonies A. Preparation of the main board Take out the single colony selected from the second round and inoculate the 96-well plate -74-016690 1316961 (70 In the continuation of the invention, each well contains 120 μl of 2xYTAG. The 96-well plate is incubated overnight at 30 ° C in a shaker. Add 40 ml of 60% glycerol to each well and - Store at 80 ° C. B. Phage ELISA assay Approximately 3 μl of cells (4A) from the native plate were seeded in a new 96-well plate containing 120 μl of 2xYTAG per well. , the new cell plate was grown at 37 ° C until OD6QQ was about 0.5. 40 ml containing M13K07 helper phage (1.2 X 101G cfu / ml Add 2xYTAG to each well and incubate the 96-well plate for 30 minutes at 37 °C without shaking, then incubate for 30 minutes with slow shaking. The plate is at 2000 rpm (Beckman CS-6R tabletop centrifuge) at 4 Centrifuge for 10 minutes at °C. Remove the supernatant from the hole and resuspend each cell pellet with 160 μl of 2 χΥΤΑΚ per well. The plates were incubated overnight at 30 ° C to allow phage to be expressed. Recombinant human MPL at a concentration of 5 μg/ml in 0.1 M carbonate buffer and pH 9.6 was applied to 96-well Maxisorp plates (NUNC) overnight at 4 °C. As a control, BSA (Sigma) was coated on a separate Maxisorp plate at a concentration of 5 μg/ml. On the next day, overnight cell cultures were centrifuged at 2000 rpm for 10 minutes at 4 °C. Transfer 20 ml of the supernatant from each well to a new 96-well plate containing 180 μl of 2% BSA/PBS solution per well. The resulting mixture was incubated at room temperature for 1 hour with shaking to block phage. At the same time, the MPL coated plates were occluded for 1 hour at room temperature using 200 μl of 2% BSA/PBS solution/well. The BSA solution was removed and each well was washed three times with PBST solution. After the last washing step, take 50 μl of the lysosome-75-016691 1316961 (71) Continuation of the invention; —J, 1» X. . Add the solution to each MPL coated plate and control In the hole of the plate, incubate for 1 hour at room temperature. The liquid was removed and each well was washed three times with PBST solution. Fifty microliters of HRP-light anti-Ml3 mAb (Amersham Pharmacia Biotech) was added to the hole of each MPL coated plate and the hole of the control plate at a dilution of 1: 15,000. The plates were incubated at room temperature. Hours. Remove the liquid again. Each well was washed three times with PBST solution. Fifty microliters of LumiGLO chemiluminescent matrix (Kirkegaard & Perry Laboratories, Gaithersburg, Maryland) was added to the caves' each well was read with a Luminoskan Ascent DLRearly machine (Labsystems, Franklin, Massachusetts). C. Sequencing of phage populations PCR was carried out using 1 μl of bacteria (4A) of each well of the autonomous plate as a template. The volume of each PCR mixture is 20 μl, which contains lx PCR buffer, two primers of 300 nM each, 5'-GTTAGCTCACTCATTAGG CAC-3, year 5'-GTACCGTAACACTGAGTTTCG-3', 200 nM dNTP, 2 mM Calcium chloride and 5 units of tag DNA polymerase (Roche Molecular Biochemicals). The following program was run using The GeneAmp PCR System 9700 (Applied Biosystem): 94. . The next 5 minutes; 40 cycles [45 seconds at 94 ° C, 45 seconds at 55 ° C, 90 seconds at 72 ° C]; 10 minutes at 72 ° C; cooled to 4 ° C. Use PCR products for QIAquick 96 PCR Purification

Kit (QIAGEN Inc.) was purified according to the manufacturer's instructions. All purified PCR products were sequenced using the primer 5'-CGGATAACAATTTCACACAGG-3 with the ABI 3770 Sequencer (Perkin Elmer) according to the manufacturer's instructions. 5. Sequence Rating 016692 -76- 1316961 r--r- (72) Description of the continuation page, correlating the peptide sequences translated from the above nucleotide sequences with ELISA data. High 〇D readings were shown in MPL coated wells, and colonies showing low 0D readings in BSA coated wells were considered candidates for further studies. Sequences that occur multiple times are also considered candidates for further research. The phage population selected based on these criteria was further identified in the ELIS A titration experiment. See Figure 9 (ELISA dose-response curve for selected phage colonies). Example 2 Preparation of peptides All peptides were prepared by existing stepwise solid phase synthesis. Merrifield (1963), J. Am. Chem. Soc. 85:2149. Steward and Young (1969), Solid Phase Peptide Synthesis. Using Fmoc-protected amino acids as constituent elements and using ABI or Symphony peptide synthesizers Construct a peptide chain. Typically, peptide synthesis begins with the use of pre-filled Wang resin to produce a peptide having a free carboxylic acid at the C-terminus (or Rink resin can be used to produce a peptide having a C-terminal guanamine function). Fmoc removal was performed using the standard piperidine procedure. A chemical reaction of a urea cation (such as HBTU) or a carbodiimide (such as DCC/HOBt) is used to facilitate coupling. The side chain protecting groups are: Glu(Ot-Bu), Asp(Ot-Bu), Ser(t-Bu), Thr(t-Bu), Arg(Pbf), Asn(Trt), Gln(Trt), His (Trt), Lys(t-Boc), Trp(t-Boc) and

Cys (Trt). The final deprotection and cleavage of all peptide-based resins was performed at room temperature using 2.5% water, 5% phenol, 2.5% triisopropyl decane and 5.2 % thiopropionate or peony Diethanol acetic acid (TFA) was carried out for 4 hours. After removal of trifluoroacetic acid, the cut peptide was lysed with cold anhydrous ether. For peptides containing disulfide bonds, use directly on crude materials, using water 016693 -77 - 1316961 -: -

(73) Description of the Invention Continued page J 15% DMSO (pH 7.5) was used to form a cyclic product. All crude peptides were purified by reverse phase HPLC and analyzed by ESI-MS and amino acid to determine the structure of the purified peptide. Example 3 Preparation of TMP-Fc Peptidomimetic Compounds Several peptides were selected to be expressed as a peptide-Fc fusion (i.e., Fc followed by the C-terminus of the peptide) (C-terminal fusion). The DNA sequence encoding the Fc region contained in human IgG1 fused to each of the TPO mimetic peptides was placed under the control of the luxPR promoter gene contained in the plastid expression vector PAMG21, as follows.

The plastid encoding the TMP1-Fc (Amgen strain, #3788) was altered to include an ApaLI site and a Xhol site for the selection of the short peptide obtained from the paired oligonucleotide. Use the primer 2396-69 to add ApaLI and XhoI to limit the S| position. The PCR system was carried out on a 3788 DNA template using Expand Long polymerase at 2396-69 and the universal 3' primer 191-24. The primer sequence was as follows: 2396-69 ACAAACAAACATATGGGTGCACAGAAAGCGGCCGCAAAAAAACT CGAGGGTGGAGGCGGTGGGGACA 191-24 GGTCATTACTGGACCGGATC The resulting PCR fragment was digested with Ndel and BsrGI, gel purified, and used as an insert. The plastid obtained from #3788 strain was also digested with Ndel and BsrGI, gel purified, and used as a carrier. The vector and insert were ligated together and the resulting ligation mixture was electroporated into GM221 cells (see below). A single colony is taken, the plastid DN A is prepared, and the DN A is sequenced. One of the resulting plasmids, 200003180, was shown to have the correct DNA sequence and was used as a vector for constructing the TMP-Fc fusion. This carrier is shown in Figure 6. -78- 016694 1316961 (74) 13⁄43⁄43⁄4 Will the plastid 200003180 be used? & 1^1 and 乂11〇1 are digested and used as a carrier. Each pair of oligonucleotides (see Figure 7) was paired to form a double strand with ApaLI and Xhol sticky ends. These molecules are ligated into a vector to produce the target fusion protein. The ApaLI and Xhol fragments of each corresponding oligonucleotide acid pair are presented in Figure 7. TMPs 1-23, 25, 26 and 28 are all expressed as C-terminal fusions. Example 4

Preparation of Fc-TMP Peptidomimetic Compounds Some peptides behave as Fc peptide fusions (ie, Fc followed by the N-terminus of the peptide) (N-terminal fusion)' will encode the plastid of Fc-TMPl (Amgen strain, #3728) Change to include an ApaLI site and an Xhol site to select for the short peptide obtained from the cooled oligonucleotide. Primers 2396-70 were designed to incorporate ApaLI and Xhol restriction enzyme sites. The PCR system was carried out on a 3728 DNA template using Expand Long polymerase at 2396-70 and the universal 5' primer 1209-85. The primer sequence was as follows: 1209-85 CGTACAGGTTTACGCAAGAAAATGG 2396-70 TTTGTTGGATCCATTACTCGAGTTTTTTTGCGGCC GCTTTCTGTGCACCACCACCTCCACCTTTAC The resulting PCR fragment was digested with BsrGI and BamHI, gel purified, and used as an insert. The plastid obtained from #3728 strain was also digested with BsrGI and BamHI, gel purified, and used as a vector. The vector and the insert were ligated together, and the resulting ligation mixture was electroporated into GM221 cells. Remove a single colony' to prepare plastid DNA and sequence the DNA. One resulting plastid, 200003182 (Figure 8), was shown to have the correct DNA sequence and was used as a vector for constructing the Fc-TMP fusion. -79- 016695 Description of Invention Continued Page 1316961 (75) Is the plastid 200003182 inserted? &!^1 and 11〇1 digested and served as a carrier. A cooled, coupled oligomer having ApaLI and Xhol's dead ends is attached to the vector to produce the target fusion. TMP20, TMP24, TMP27, TMP29 and TMP30 were produced in this manner as N-terminal fusions. Transformation Each of the above-described conjugates was transformed into host species GM22 1 by electroporation as described below. The selection is screened for the ability to produce recombinant protein products and possess the ability to fuse with the correct nucleotide sequence. The PAMG21 performance plastid PAMG21 is available from ATCC under accession number 98113 and has a retention period of July 24, 1996. GM221 (Amgen host strain #2596)

Amgen host strain #2596 is the E. coli K-12 strain which has been modified to include the temperature-sensitive lambda inhibitor cI857s7 in the early ebg region and the lacIQ inhibitor in the late ebg region (68 minutes). The presence of this rain suppressor gene allows the host to be used with a variety of expression systems, although the two inhibitors are independent of luxPj^^ performance. Untransformed hosts do not have antibiotic resistance. The ribosome binding site of the cI857s7 gene has been modified to include an enhanced RBS. It is inserted into the ebg operon, which is between the nucleotide positions 1170 and 1411, and is numbered 64441〇1)_ Ba in the 〇61^&1^ accession number, in which the intervention is deleted. Ebg sequence. The construct was delivered to the chromosome using recombinant phage, which was MMebg-cI857s7 enhanced RBS #4 into F'tet/393. After recombination and resolution, only the above chromosomal insert remains in the cell. It was renamed to F'tet/GMIOl. The F'tet/GMIOl is then modified into the ebg operon between the nucleotide positions 2493 and 293 7 to be modified into the 'Genΐ' ank registration number numbered M64441Gb_Ba, in which the intervening ebg is deleted. sequence. This construct was delivered to the chromosome using a recombinant phage of AGebg-LacIQ#5 into F'tet/GMIO1. After recombination and resolution, only the above-described dye-chromosome insert remains in the cell. Rename it to F'tet/GM221. The F'tet gene epithelium was titered from the strain at a concentration of 25 μg/ml in LB using t»丫 precipitated orange. The recovered strain was identified as a tetracycline-sensitive strain and stored as GM221. Performance A culture of GM221 that exhibited each fusion was placed at 37. (: Growth in Luria Broth medium. The step of inducing gene product expression from iuxPR promoter is through the addition of the synthetic autoinducer N_(3_ketohexylidene)_DL-homserine to the final concentration in the medium. 2 〇 nanograms/ml' and incubated for another 3 hours at 371 and ligated. After 3 hours, the bacterial culture was observed by microscopy for the presence of inclusion bodies, and then centrifuged to collect them. The refractive inclusion bodies were induced. It was observed in the culture that this indicated that the fused protein was most likely to be insoluble in E. coli. The cells were lysed and resuspended in Laemmli sample buffer containing 10% cold-mercaptoethanol. SDS-PAGE analysis. Observed for each protein 13 - a strong Coomassie staining band of appropriate size (approximately 30 -81-016697 (77) 1316961 kDa). Example 5 Purification of the peptide compound in water It was ruptured by high pressure homogenization (2-way at 14,000 PSI) (1/10) and the inclusion bodies were harvested by centrifugation (4, 2 rpm in J-6B '1 hour). The body was dissolved in 6 μ of pulse, 50 mM Tris, 8 mM DTT at pH 8.7 ' for 1 hour. The dissolved mixture was diluted 20-fold to 2 脲 urea, 50 mM Tris, 160 mM arginine, 3 mM cysteine' pH was 8.5. The mixture was stirred overnight at low temperature. The mixture was then concentrated approximately 10 times by ultrafiltration. Then 10 mM Tris, 1.5 Μ was used. The urea is diluted 3 times and the pH is 9. The pH of the mixture is then adjusted to pH 5 with acetic acid. The precipitate is removed by centrifugation and loaded onto the SP-Sepharose Fast Flow column. The column was equilibrated with 20 mM NaAc, 100 mM sodium chloride, pH 5 (10 mg/ml protein loading, room temperature). Using 20 column volumes, 1 mM HCl in the same buffer. The protein was eluted by sodium to 500 mM sodium chloride gradient. The combined fraction obtained from the tube was diluted 3 times and loaded onto SP-Sepharose HP column '20 mM NaAc, 150 mM cesium chloride, pjj 5 in (10 mg/ml protein loading, at room temperature). Use 20 tubes to get the main volume in the same buffer The protein was eluted with a gradient of 150 mM sodium chloride to 400 mM chloride. The peaks were combined and filtered. Example 6 Peptide affinity binding study Using BIACORE 3000, experiments were performed at room temperature to determine several Tlvlp-82-1316961 ( 78) Cross-Ming Description Continued Page 1 The peptide's binding affinity (ΤΜΡ1-ΤΜΡ23). Hu-mpl was immobilized on the surface of the sensor wafer (CM5) using an amine coupling procedure (activated with NHS/EDC and blocked with ethanolamine). A 0.78 nM to 100 nM TMP peptide was injected onto the Hu-mpl surface. The BIACORE working buffer was PBS plus 0.005% surfactant P20. The sample was also injected onto a blank surface as a control. The experimental data was analyzed using the BIAEVALUATION 3.1 suite of software. As discussed above, in order to better mimic the phage environment in which the peptide is selected, and in order to eliminate the charged amine and carboxyl-ends of the 18 amino acid preferred peptides (TMP2-TMP30) from the receptor, Two amino acids "caps are added to each carboxy terminus and amine terminus of each peptide: glutamine-cysteine (QC) to the amine end, and histidine-silyl group (HS) to the carboxy terminus, changing the length of each peptide to 22 amino acids. Since the affinity of the peptide is known to increase with the length of the peptide, it will also be labeled with 14 amino groups. The acid sequence (SEQ ID NO 1) was increased to a total of 22 amino acids. However, the bioactive region of each peptide remained unchanged as shown in bold in the table below. TMP No. peptide sequence KD(nM) relative 1 TMP in affinity TMP1 SAQGIEGPTLRQWLAARALETV 5.40 _ TMP2 QGGAREGPTLRQWLEWVRVGHS 1.60 3.38 TMP3 QGRDLDGPTLRQWLPLPSVQHS 45.00 0.12 TMP4 QGALRDGPTLKQWLEYRRQAHS 0.86 6.28 TMP5 QGARQEGPTLKEWLFWVRMGHS 6.66 0.81 TMP6 QGEALLGPTLREWLAWRRAQHS 0.37 14.59 TMP7 QGMARDGPTLREWLRTYRMMHS 1.20 4.50 TMP8 QGWMPEGPTLKQWLFHGRGQH S 23.20 0.23 016699 1316961 invention is described $ p _ TMP9 QGHIREGPTLRQWLVALRMVHS 1.67 3.23 TMP10 QGQLGHGPTLRQWLSWYRGMHS 1.22 4.43 TMP11 QGELRQGPTLHEWLQHLASKHS 35.90 0.15 TMP12 QGVGIEGPTLRQWLAQRLNPHS 5.20 1.04 TMP13 QGWSRDGPTLREWLAWRAYGHS 4.44 1.22 TMP14 QGAVPQGPTLKQWLLWRRCAHS 0.88 6.14 TMP15 QGRIREGPTLKEWLAQRRGFHS 1.03 5.24 TMP16 QGRFAEGPTLREWLEQRKLVHS 6.58 0.82 TMP17 QGDRFQGPTLREWLAAIRSVHS 12.90 0.42 TMP18 QGAGREGPTLREWLNMRVWQHS 12.80 0.42 TMP19 QGALQEGPTLRQWLGWGQWGHS 78.50 0.07 TMP20 QGYCDEGPTLKQWLVCLGLQHS 0.56 9.64 TMP21 QGWCKEGPTLREWLRWGFLCHS 1.53 3.53 TMP22 QGCSSGGPTLREWLQCRRMQHS <0.1 >54 TMP23 QGCSWGGPTLKQWLQCVRAKHS <0.1 >54 Example 7 Peptide Bioactivity Studies Cell-based assays were used to determine the bioactivity of peptide TMP1-TMP23 . Murine 32D cell proliferation assays include murine 32D cells transfected with human mpl receptor. The following results are reported relative to TMP1. The CD61 cell assay involves the use of pro-human CD34+ cells cultured for several days in the presence of the peptide TMP1-TMP23. These cells are then classified to determine the percentage of cells that can exhibit a megakaryocyte-specific marker (CD61) on the cell surface. Stimulation of 016700-84-1316961 (80) in a dose-dependent manner using active compounds. [Inventive Notes] These platelet precursor cells are present when red blood cell precursor (CD36+) and neutrophil precursor (CD 15 + ) The marker is still at baseline. The qualitative results of the CD61 cell assay are shown below, which present an average of three different concentrations. Peptide mouse 32D cell proliferation assay (relative to TMP1) CD61 cell assay (relative to TMP1) TMP01 100% -/+ TMP02 290% + TMP03 39% ++ TMP04 42% TMP05 85% ++ TMP06 569% ++ TMP07 289 % ++ TMP08 39% + TMP09 2% TMP10 12% . TMP11 21% . TMP12 10% TMP13 328% ++ TMP14 635% +++ TMP15 35% TMP16 32% + TMP17 21% . TMP18 337% ++ TMP19 27 % + -85- 016701 1316961 (81) TMP20 can't detect _/+ TMP21 312% -/+ TMP22 can't detect TMP23 can't detect +++ Example 8 Peptide binding study on BIAcore with direct binding analysis, The binding activity of several TMP peptides to hu-MPL was tested. The experiment was carried out at 25 ° C using BIAcore 2000 (BIACORE Inc.). The running buffer was 5% plus surfactant surfactant P20. Recombinant protein G (Pierce 21193ZZ) was immobilized on the surface of a CM5 wafer using an amine coupling procedure (activated with NHS/EDC and blocked with ethanolamine) to capture the TMP peptide to about 400 RU. Recombinant hu-MPL (Lot 273 15-53) was serially diluted from 1 #Μ to 0.15 nM in sample buffer (PBS plus 0.005% surfactant P20 and 100 μg/ml BSA), then 50 microliters per minute was injected on the surface of the captured peptide body for 3 minutes. The rhu-MPL sample was also injected onto the blank protein G surface to subtract any non-specific binding background. The protein G surface was sequentially injected with 100 μl of ImmunoPure IgG Lysis Buffer (Pierce 21009ZZ, pH 2) and 100 μl of 8 mM Glycine, pH 1.5, 1 at 50 μL/min between two cycles. The gasified sodium of Μ is regenerated. The binding affinity (KD) of the peptide to rhu-MPL was determined by nonlinear regression analysis of the data using BIAevaluation 3.1 software package (BIACORE Inc.). The results are summarized as follows: -86- 01Θ702 1316961 Issue b month description Continued page 1 Multi-peptide (TMP-Fc) ka(l/Ms) kd(l/s) Kd(M) TMP20-FC 5.06 x 104 7.34 χ ΙΟ' 3 1.45 χ ΙΟ'7 Fc-TMP24 4.01 χ 104 8_75 χ 10-3 2.18 χ ΙΟ'7 TMP25-Fc 2.35 χ 104 1.40 χ ΚΤ3 5.97 χ ΙΟ'8 TMP26-Fc 2.58 χ ΙΟ4 5.72 χ ΙΟ'3 2.22 χ ΙΟ '7 Fc-TMP27 1.3 χ ΙΟ5 8.42 χ ΙΟ*3 6.49 χ 8'8 TMP28-FC 6.78 χ ΙΟ4 2.52 χ 1 (Τ2 3.71 χ ΙΟ'7 Example 9 Peptide activity assay. Put human CD34+ cells in number Cultured in the presence of a TMP-Fc fusion protein for several days. These cells were then sorted to determine the percentage of cells that exhibited a megakaryocyte-specific marker (CD61) on the cell surface. These compounds were stimulated in a dose-dependent manner using the active compound. At the onset of platelet precursor cells, markers of red blood cell precursor (CD36 + ) (not shown) and neutrophil precursor (CD 15 + ) (not shown) are still at baseline. See Figures 10, 11 and 12 ( CD61 cell assay). Example 10 In vivo activity Using normal female BDF1 mice, approximately 10-12 weeks old, live In the internal activity study, mice were injected simultaneously for bolus treatment. The subcutaneous injection was delivered in a volume of 0.2 ml. The compound was diluted in PBS plus 0.1% BSA. All experiments included a control group. Diluent-treated label "carrier π. -87- 016703 1316961 (83) For each group of ten mice on day 0, the start time of the two groups was 4 days, and the total number was 20 mice per group. The rats were bled at each time point and the mice were bled at least three times per week. The mice were anesthetized with isoflurane and the total volume of blood was 140-160 microliters by burrs of the eyelids. The blood was placed at Technicon H1E. The mouse blood analysis software is run on the blood analyzer for counting. The measured parameters are white blood cells, red blood cells, hematocrit, heme, platelets, neutrophils. See Figures 13 and 14. The present invention has now been fully described, It will be apparent to those skilled in the art that various changes and modifications can be made thereto without departing from the spirit and scope of the invention as described herein. 88-016704 1316961 Sequence Listing <11〇><120>Peptide-producing peptide and related compound <130>A-750 <140> TW 091123464 <141> 2002-10-11 <150> US 60/328,666 <151> 2001-10- 11 <160> 199 <170> Paten tin version 3.1 <210> 1 <211> 18 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400 > 1

Gin Gly lie Glu Gly Pro Thr Leu Arg Gin Trp Leu Ala Ala Arg Ala 1 5 10 15

Leu Glu <210> 2 <2il> 18 <212> PRT <213> artificial sequence <220><223> synthetic peptide sequence <400> 2 .

Gly Ala Arg Glu Gly Pro Thr Leu Arg Gin Trp Leu Glu Trp Val Arg '1 5 10 15

Val Gly <210> 3 016705 1316961 <211> 18 <212> PRT <213> artificial sequence <220><223> synthetic winning sequence <400> 3

Arg Asp Leu Asp Gly Pro Thr Leu Arg Gin Trp Leu Pro Leu Pro Ser 15 10 15

Val Gin <210> 4 <211> 18 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Ala Leu Arg Asp Gly Pro Thr Leu Lys Gin Trp Leu Glu Tyr Arg Arg 15 10 15

Gin Ala <210> 5 <211> 18 <212> PRT <213> Artificial sequence <220><223> Synthetic peptide sequence <400>

Ala Arg Gin Glu Gly Pro Thr Leu Lys Glu Trp Leu Phe Trp Val Arg 15 10 15 .Met Gly <210〉 6

01670S 1316961 <211> 18 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Glu Ala Leu Leu Gly Pro Thr Leu Arg Glu Trp Leu Ala Trp Arg Arg 15 10 15

Ala Gin <210> 7 <211> 18 <212> PRT <213> artificial sequence <220><223> synthetic winning sequence <400>

Met Ala Arg Asp Gly Pro Thr Leu Arg Glu Trp Leu Arg Thr Tyr Arg 15 10 15

Met Met <210> 8 <211> 18 <212> PRT <2i3> artificial sequence <220><223> synthetic peptide sequence <400>

Trp Met Pro Glu Gly Pro Thr Leu Lys Gin Trp Leu Phe His Gly Arg 15 10 15

Gly Gin <210> 9 -3- 016707 1316961 <211> 18 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400> 9

His lie Arg Glu Gly Pro Thr Leu Arg Gin Trp Leu Val Ala Leu Arg 15 10 15

Met Val <210> 10 <211> 18 <212> PRT <213>Artificial sequence <220><223> Synthesis peptide sequence <400>

Gin Leu Gly His Gly Pro Thr Leu Arg Gin Trp Leu Ser Trp Tyr Arg 15 10 15

Gly Met <210> 11 <211> 18 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Glu Leu Arg Gin Gly Pro Thr Leu His Glu Trp Leu Gin His Leu Ala 15 10 15

Ser Lys <210> 12 016708 1316961 <211> 18 <212> PRT <213>Artificial sequence <220><223>Synthesis peptide sequence <400>

Val Gly lie Glu Gly Pro Thr Leu Axg Gin Trp Leu Ala Gin Arg Leu 15 10 15

Asn Pro <210> 13 <211> 18 <212> PRT <213> Artificial sequence <220><223> Synthetic peptide sequence <400>

Trp Ser Arg Asp Gly Pro Thr Leu Arg Glu Trp Leu Ala Trp Arg Ala 15 10 15

Val Gly <210> 14 <211> 18 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400> 14

Ala Val Pro Gin Gly Pro Thr Leu Lys Gin Trp Leu Leu Trp Arg Arg 15 10 15

Cys Ala <210> 15 Π1β7Πύ 1316961 <211> 18 <212> PRT <213> Artificial sequence <220><223> Synthetic peptide sequence <400>

Arg lie Arg Glu Gly Pro Thr Leu Lys Glu Trp Leu Ala Gin Arg Arg 15 10 15

Gly Phe <210> 16 <211> 18 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Arg Phe Ala Glu Gly Pro Thr Leu Arg Glu Trp Leu Glu Gin Arg Lys 15 10 15

Leu Val <210> 17 <211> 18 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Asp Arg Phe Gin Gly Pro Thr Leu Arg Glu Trp Leu Ala Ala lie Arg 15 10 15

Ser Val <210> 18 016710 -6- 1316961 <211> 18 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Ala Gly Arg Glu Gly Pro Thi* Leu Arg Glu Trp Leu Asn Met Arg Val 15 10 15

Trp Gin <210> 19 <211> 18 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Ala Leu Gin Glu Gly Pro Thr Leu Arg Gin Trp Leu Gly Trp Gly Gin 15 10 15

Trp Gly <210> 20 <211> 18 <212> PRT <213>Artificial sequence <220><223> Synthetic skin sequence <400> 20

Tyr Cys Asp Glu Gly Pro Thr Leu Lys Gin Trp Leu Val Cys Leu Gly 15 10 15

Leu Gin <210> 21 016711 1316961 <211> 18 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Trp Cys Lys Glu Gly Pro Thr Leu Arg Glu Trp Leu Axg Trp Gly Phe 15 10 15

Leu Cys <210> 22 <211> 18 <212> PRT <213> artificial sequence <220><223> synthetic peptide sequence <400>

Cys Ser Ser Gly Gly Pro Thr Leu Arg Glu Trp Leu Gin Cys Arg Arg 15 10 15

Met Gin <210> 23 <211> 18 <212>PRT <2i3> artificial sequence <220><223> synthetic peptide sequence <400>

Cys Ser Trp Gly Gly Pro Thr Leu Lys Gin Trp Leu Gin Cys Val Arg 1 5 10 15

Ala Lys <210> 24 016712 1316961 <211> 18 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Cys Gin Leu Gly Gly Pro Thr Leu Arg Glu Trp Leu Ala Cys Arg Leu 15 10 15

Gly Ala <210> 25 <211> 18 <212>PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Cys Trp Glu Gly Gly Pro Thr Leu Lys Glu Trp Leu Gin Cys Leu Val 15 10 15

Glu Arg <210> 26 <211> 18 <212> PRT <213>Artificial sequence<220><223> Synthetic peptide sequence <400>

Cys Arg Gly Gly Gly Pro Thr Leu His Gin Trp Leu Ser Cys Phe Arg 15 10 15

Trp Gin <210> 27 016713 9- 1316961 <211> 18 <212> PRT <213>Artificial sequence <220><223><223> Synthetic peptide sequence <400>

Cys Arg Asp Gly Gly Pro Thr Leu Arg Gin Trp Leu Ala Cys Leu Gin 15 10 15

Gin Lys <210> 28 <211> 18 <212> PRT <2i3> artificial sequence <220><223> synthetic peptide sequence <400>

Glu Leu Arg Ser Gly Pro Thr Leu Lys Glu Trp Leu Val Trp Arg Leu 15 10 15

Ala Gin <210> 29 <211> 18 <212> PRT <213> artificial sequence <220><223> synthetic peptide sequence <400>

Gly Cys Arg Ser Gly Pro Thr Leu Arg Glu Trp Leu Ala Cys Arg Glu 15 10 15

Val Gin <210> 30 0.^6711 1316961 <211> 18 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400> 30

Thr Cys Glu Gin Gly Pro Thr Leu Arg Gin Trp Leu Leu Cys Arg Gin 15 10 15

Gly Arg <210> 31 <211> 684 <212> DNA <213> Artificial sequence - <220><223> Synthetic peptide sequence <220><221> CDS <222> 1)..(684) <223><400> 31 atg gac aaa act cac aca tgt cca cct tgt cca get cog gaa etc ctg 48 Met Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 15 10 15 ggg gga ccg tea gtc ttc etc ttc ccc cca aaa ccc aag gac acc etc 96 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 20 25 30 atg ate tcc egg acc cct gag gtc aca tgc gtg gtg Gtg gac gtg age 144 Met He Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 35 40 45 cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg gag 192 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 50 55 60 gtg cat aat gcc aag aca aag ccg egg gag gag cag tac aac age aeg 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr 65 70 75 80 tac cgt gtg Gtc age gtc etc acc gtc ctg cac cag gac tgg ctg aat 288 Tyr Arg Val Val Ser Val Le u Thr Val Leu His Gin Asp Trp Leu Asn 85 90 95 016715 -11 - 1316961 ggc aag gag tac aag tgc aag gtc tcc aac aaa gcc etc cca gee ccc 336 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asa Lys Ala Leu Pro Ala Pro 100 105 110 ate gag aaa acc ate tec aaa gcc aaa ggg cag ccc ega gaa cca cag 384 lie Glu Lys Thr lie Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin 115 120 125 gtg tac acc ctg ccc cca tec egg gat gag Ctg acc aag aac cag gtc 432 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gin Val 130 135 140 age ctg acc tgc ctg gtc aaa ggc ttc tat ccc age gac ate gcc gtg 480 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val 145 150 155 160 gag tgg gag age aat ggg cag ccg gag aac aac tac aag acc aeg cot 528 Glu Trp Glu Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro 165 170 175 ccc Gg tec gac tec gac ggc tec ttc ttc etc tac. age aag etc acc 576 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 180 185 190 gtg gac aag age agg tgg cag cag ggg aac gtc ttc tea tgc Tec gtg 624 Val Asp Lys Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val 195 200 205 atg cat gag get ctg cac aac cac tac aeg cag aag age etc tec ctg 672 Met His Glu Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu 210 215 220 tet ccg ggt aaa 684

Ser Pro Gly Lys 225 <210> 32 <211> 228 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Met Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 15 10 15

Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 20 25 30 016716 12- 1316961

Met lie Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 35 40 45

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 50 55 60

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr 65 70 75 80

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn 85 90 95

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 100 105 _ 110 lie Glu Lys Thr lie Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin 115 120 125

Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gin Val 130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val 145 150 155 160

Glu Trp Glu Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro 165 170 175 'Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 180 185 190

Val Asp Lys Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val 195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu 210 215 220

Ser Pro Gly Lys 225 01671? -13 - 1316961 <210> 33 <211> 835 <212> DNA <213>Artificial sequence <220><223> Synthetic peptide sequence <220>221> CDS <222>(60),.(791) <223><400> 33 tagtcgatta atcgatttga ttctagattt gttttaacta attaaaggag gaataacat 59 atg ggt gca cag aaa gcg gcc gca aaa aaa etc gag ggt gga ggc ggt 107 Met Gly Ala Gin Lys Ala Ala Ala Lys Lys Leu Glu Gly Gly Gly Gly 15 10 15 ggg gac aaa act cac aca tgt cca cct tgc cca gca cct gaa etc ctg 155 Gly Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 20 25 30 ggg gga ccg tea gtt ttc etc ttc ccc cca aaa ccc aag gac acc etc 203 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 35 40 45 atg ate tee egg acc cct gag gtc aca tgc gtg Gtg gtg gac gtg age 251 Met lie Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 50 55 60 cac gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg gag 299 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 65 70 75 80 gtg cat aat gcc aag aca aag ccg egg gag gag cag tac aac age aeg 347 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Tyr Asn Ser Thr 85 90 95 tac cgt gtg gtc age gtc etc acc gtc ctg cac Cg gac tgg ctg aat 395 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn 100 105 110 ggc aag gag tac aag tgc aag gtc tee aac aaa gcc etc cca gcc ccc 443 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 115 120 125 ate gag aaa acc ate tee aaa gcc aaa ggg cag ccc ega gaa cca cag 491 lie Glu Lys Thr lie Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin 130 135 140 gtg tac acc Ctg ccc cca tee egg gat gag ctg acc aag aac cag gtc 539 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gin Val 016718 -14- 1316961 145 150 155 160 age ctg acc tgc ctg gtc aaa ggc ttc tat ccc Age gac ate gee gtg 587 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val 165 170 175 gag tgg gag age aat ggg cag ccg gag aac aac tac aag acc aeg cct 635 Glu Trp Glu Ser Asn Gly Gin Pro Glu As n Asn Tyr Lys Thr Thr Pro 180 185 190 ccc gtg ctg gac tee gac ggc tee ttc ttc etc tac age aag etc acc 683 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 195 200 205 gtg gac aag age Agg tgg cag cag ggg aac gtc ttc tea tgc tee gtg 731 Val Asp Lys Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val 210 215 220 atg cat gag get ctg cac aac cac tac aeg cag aag age etc tee ctg 779 Met His Glu Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu 225 230 ' 235 240 tet ccg ggt aaa taatggatcc gcggaaagaa gaagaagaag aagaaagccc gaaa 835 Ser Pro Gly Lys <210> 34 <211> 244 <212> PRT <;213>Artificial sequence <220> <223> Synthetic peptide sequence <400> 34

Met Gly Ala Gin Lys Ala Ala Ala Lys Lys Leu Glu Gly Gly Gly Gly 15 10 15

Gly Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 20 30

Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 35 40 45

Met lie Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 50 55 60

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 016719 -15 - 1316961 65 70 75 80

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr 85 90 95

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn 100 105 110

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 115 120 125 lie Glu Lys Thr lie Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin 130. 135 140

Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gin Val 145 150 _ 155 160

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val 165 170 175

Glu Trp Glu Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro 180 185 190

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 195 200 205

Val Asp Lys Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val 210 215 220

Met His Glu Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu 225 230 235 240

Ser Pro Gly Lys <210> 35 <211> 66 <212> DNA <213>Artificial sequence <220><223> Synthetic peptide sequence 016720 -16 - 1316961 <220> <221> CDS <222> (4)..(66) <223><400> 35 cat atg ggt gca cag ggt ate gaa ggt ccg act ctg cgt cag tgg ctg 48 Met Gly Ala Gin Gly lie Glu Gly Pro Thr Leu Arg Gin Trp Leu 15 10 15 get get cgt get etc gag 66

Ala Ala Arg Ala Leu Glu 20 <210> 36 <211> 21 <212>PRT <213> artificial sequence <220>, <223> synthetic peptide sequence <400>

Met Gly Ala Gin Gly lie Glu Gly Pro Thr Leu Arg Gin Trp Leu Ala 15 10 15

Ala Arg Ala Leu Glu 20 <210> 37 <211> 71 <212>PRT <213> Artificial shore column <220> . <223> Synthetic peptide sequence <400>

Thr Gly Cys Ala Cys Ala Ala Gly Gly Thr Gly Gly Ala Gly Cys Ala 1 5 10 15

Cys Gly Thr Gly Ala Ala Gly Gly Ala Cys Cys Ala Ala Cys Thr Cys 20 25 30

Thr Thr Cys Gly Thr Cys Ala Ala Thr Gly Gly Cys Thr Thr Gly Ala 35 40 45 -17- 1316961

Ala Thr Gly Gly Gly Thr Thr Cys Gly Thr Gly Thr Thr Gly Gly Thr 50 55 60

Cys Ala Thr Thr Cys Thr Cys 65 70 <210> 38 <211> 71 <212> PRT <213> Artificial sequence <220> 1 <223> Synthetic peptide sequence <400>

Thr Cys Gly Ala Gly Ala Gty Ala Ala Thr Gly Ala Cys Cys Ala Ala 15 10 15

Cys Ala Cys Gly Ala Ala Cys Cys Cys Ala Thr Thr Cys Ala Ala Gly 20 25 30

Cys Cys Ala Thr Thr Gly Ala Cys Gly Ala Ala Gly Ala Gly Thr Thr 35 40 45

Gly Gly Thr Cys Cys Thr Thr Cys Ala Cys Gly Thr Gly Cys Thr Cys 50 55 60

Cys Ala Cys Cys Thr Thr Gly 65 70 <210> 39 <211> 77 <212> DNA <213>Artificial Sequence <220><223> Synthetic Peptide Sequence <220><221> CDS <222> (3)..(77) <223><400> 39 016722 18- 1316961 gt gca caa ggt gga gca cgt gaa gga cca act ctt cgt caa tgg ctt 47 Ala Gin Gly Gly Ala Arg Glu Gly Pro Thr Leu Arg Gin Trp Leu 15 10 15 gaa tgg gtt cgt gtt ggt cat tct etc gag ΊΊ

Glu Trp Val Arg Val Gly His Ser Leu Glu 20 25 <210> 40 <211> 25 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400> 40

Ala Gin Gly Gly Ala Arg Glu Gly Pro Thr Leu Arg Gin Trp Leu Glu 1 5 10. 15

Trp Val Arg Val Gly His Ser Leu Glu 20 25 <210> 41 <211> 71 <212> PRT <213>Artificial Sequence <220><223> Synthetic Skin Sequence <400>

Thr Gly Cys Ala Cys Ala Ala Gly Gly Ala Cys Gly Thr Gly Ala Thr 15 10 15

Cys Thr Thr Gly Ala Thr Gly Gly Thr Cys Cys Ala Ala Cys Thr Cys 20 25 30

Thr Thr Cys Gly Thr Cys Ala Ala Thr Gly Gly Cys Thr Thr Cys Cys 35 40 45

Ala Cys Thr Thr Cys Cys Ala Thr Cys Thr Gly Thr Thr Cys Ala Ala 50 55 60

Cys Ala Thr Thr Cys Thr Cys 016723 19- 1316961 65 70 <210> 42 <211> 71 <212> PRT <213>Artificial Sequence<220><223> Synthetic peptide sequence <400> 42

Thr Cys Gly Ala Gly Ala Gly Ala Ala Thr Gly Thr Thr Gly Ala Ala 15 10 15

Cys Ala Gly Ala Thr Gly Gly Ala Ala Gly Thr Gly Gly Ala Ala Gly 20 25 30

Cys Cys Ala Thr Thr Gly Ala Cys Gly Ala Ala Gly Ala Gly Thr Thr 35 40 45

Gly Gly Ala Cys Cys Ala Thr Cys Ala Ala Gly Ala Thr Cys Ala Cys 50 55 60

Gly Thr Cys Cys Thr Thr Gly 65 70 <210> 43 <211> 77 <212> DNA <213>Artificial Sequence <220><223> Synthetic Peptide Sequence <220><221> CDS <222> (3)..(77) <223><400> 43 gt gca caa gga cgt gat ctt gat ggt cca act ctt cgt caa tgg ctt 47 Ala Gin Gly Arg Asp Leu Asp Gly Pi*o Thr Leu Arg Gin Trp Leu 1 5 10 15 cca ctt cca tct gtt caa cat tct etc gag 77

Pro Leu Pro Ser Val Gin His Ser Leu Glu 20 25 016724 20- 1316961 <210> 44 <211> 25 <212> PRT <213>Artificial Sequence<220><223> Synthetic peptide sequence<223>;400> 44

Ala Gin Gly Arg Asp Leu Asp Gly Pro Thr Leu Arg Gin Trp Leu Pro 15 10 15

Leu Pro Ser Val Gin His Ser Leu Glu 20 25 <210> 45 <211> 71 <212> PRT <213>Artificial Sequence <220><223> Synthetic Skin Sequence <400> 45

Thr Gly Cys Ala Cys Ala Ala Gly Gly Ala Gly Cys Thr Thr Thr Ala 15 10 15

Cys Gly Thr Gly Ala Thr Gly Gly Thr Cys Cys Ala Ala Cys Thr Cys 20 25 30

Thr Thr Ala Ala Ala Cys Ala Ala Thr Gly Gly Thr Thr Ala Gly Ala 35 40 45

Ala Thr Ala Thr Cys Gly Thr Cys Gly Thr Cys Ala Ala Gly Cys Thr 50 55 60

Cys Ala Thr Thr Cys Ala Cys 65 70 <210> 46 <211> 71 <212> PRT <213> Artificial sequence 016725 1316961 <220><223> Synthetic peptide sequence <400>

Thr Cys Gly Ala Gly Thr Gly Ala Ala Thr Gly Ala Gly Cys Thr Thr 15 10 15

Gly Ala Cys Gly Ala Cys Gly Ala Thr Ala Thr Thr Cys Thr Ala Ala 20 25 30

Cys Cys Ala Thr Thr Gly Thr Thr Thr Ala Ala Gly Ala Gly Thr Thr 35 40 45

Gly Gly Ala Cys Cys Ala Thr Cys Ala Cys Gly Thr Ala Ala Ala Gly 50 55 60

Cys Thr Cys Cys Thr Thr Gly 65 70 <210> 47 <211> 77 <212> DNA <213>Artificial Sequence <220><223> Synthetic Peptide Sequence <220><221> CDS <222>(3)., (77) <223><400> 47 gt gca caa gga get tta cgt gat ggt cca act ett aaa caa tgg tta 47 Ala Gin Gly Ala Leu Arg Asp Gly Pro Thr Leu Lys Gin Trp Leu 15 10 15 gaa tat cgt cgt caa get cat tea etc gag 77

Glu Tyr Arg Arg Gin Ala His Ser Leu Glu 20 25 <210> 48 <211> 25 <212> PRT <213>Artificial Sequence 016726 -22 - 1316961 <220><223> Synthetic peptide sequence <400> 48

Ala Gin Gly Ala Leu Arg Asp Gly Pro Thr Leu Lys Gin Trp Leu Glu 15 10 15

Tyr Arg Arg Gin Ala His Ser Leu Glu 20 25 <210> 49 <211> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Thr Gly Cys Ala Cys Ala Ala Gly Gly Ala Gly Cys Ala Cys Gly Thr 15 10 15

Cys Ala Ala Gly Ala Ala Gly Gly Ala Cys Cys Ala Ala Cys Thr Cys 20 25 30

Thr Thr Ala Ala Ala Gly Ala Ala Thr Gly Gly Thr Thr Ala Thr Thr 35 40 45

Thr Thr Gly Gly Gly Thr Thr Cys Gly Thr Ala Thr Gly Gly Gly Thr 50 55 60

Cys Ala Thr Thr Cys Ala Cys 65 70 <210> 50 <211> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Thr Cys Gly Ala Gly Thr Gly Ala Ala Thr Gly Ala Cys Cys Cys Ala 018727 -23- 1316961 15 10 15

Thr Ala Cys Gly Ala Ala Cys Cys Cys Ala xA.la Ala Ala Thr Ala Ala 20 25 30

Cys Cys Ala Thr Thr Cys Thr Thr Thr Ala Ala Gly Ala Gly Thr Thr 35 40 45

Gly Gly Thr Cys Cys Thr Thr Cys Thr Thr Gly Ala Cys Gly Thr Gly 50 55 60

Cys Thr Cys Cys Thr Thr Gly 65 70 <210> 51 <211> 77 <212> DNA <213>Artificial Sequence <220><223> Synthetic Peptide Sequence <220><221> CDS <222> 0)..(77) <223><400> 51 gt gca caa gga gca cgt caa gaa gga cca act ctt aaa gaa tgg tta 47 Ala Gin Gly Ala Arg Gin Glu Gly Pro Thr Leu Lys Glu Trp Leu 1 5 10 15 ttt tgg gtt cgt atg ggt cat tea etc gag 77

"health sequence <220><223&gt

Ala Gin Gly Ala Arg Gin Glu Gly Pro Thr Leu Lys Glu Trp Leu Phe 15 10 15 m β7〇β -24- 1316961

Trp Val Arg Met Gly His Ser Leu Glu 20 25

<210> 53 <211> 71 <212> PRT <213> Artificial sequence <220><223> Synthetic peptide sequence <400>

Thr Gly Cys Ala Cys Ala Ala Gly Gly Ala Gly Ala Ala Gly Cys Thr 15 10 15

Thr Thr Ala Thr Thr Ala Gly Gly Thr Cys Cys Ala Ala Cys Thr Thr 20 25 30

Thr Ala Cys Gly Thr Gly Ala Ala Thr Gly Gly Cys Thr Thr Gly Cys 35 40 45

Thr Thr Gly Gly Cys Gly Thr Cys Gly Thr Gly Cys Ala Cys Ala Ala 50 55 60

Cys Ala Thr Thr Cys Thr Cys 65 70 <210> 54 <211> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Thr Cys Gly Ala Gly Ala Gly Ala Ala Thr Gly Thr Thr Gly Thr Gly 15 10 15

Cys Ala Cys Gly Ala Cys Gly Cys Cys Ala Ala Gly Cys Ala Ala Gly 20 25 30 r\ λ η Λ -25 - 1316961

Cys Cys Ala Thr Thr Cys Ala Cys Gly Thr Ala Ala Ala Gly Thr Thr 35 40 45

Gly Gly Ala Cys Cys Thr Ala Ala Thr Ala Ala Ala Gly Cys Thr Thr 50 55 60

Cys Thr Cys Cys Thr Thr Gly 65 70 <210> 55 <211> 77 <212> DNA <213>Artificial Sequence <220><223> Synthetic peptide sequence <220><221> CDS <222> (3)..(77) <223><400> 55 gt gca caa gga gaa get tta tta ggt cca act tta cgt gaa tgg ett 47 Ala Gin Gly Glu Ala Leu Leu Gly Pro Thr Leu Arg Glu Trp Leu 15 10 15 get tgg cgt cgt gca caa cat tet etc gag 77

Ala Trp Arg Arg Ala Gin His Ser Leu Glu 20 25 <210> 56 <211> 25 <212> PRT <213> Artificial sequence <220><223> Synthetic peptide sequence <400>

Ala Gin Gly Glu Ala Leu Leu Gly Pro Thr Leu Arg Glu Trp Leu Ala 15 10 15

Trp Arg Arg Ala Gin His Ser Leu Glu 20 25 <210> 57 016730 -26- 1316961 <211> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <223>;400> 57

Thr Gly Cys Ala Cys Ala Ala Gly Gly Thr Ala Thr Gly Gly Cys Ala 15 10 15

Cys Gly Thr Gly Ala Thr Gly Gly Thr Cys Cys Ala Ala Cys Thr Cys 20 25 30

Thr Thr Cys Gly Thr Gly Ala Ala Thr Gly Gly Cys Thr Thr Cys Gly 35 40 45

Thr Ala Cys Thr Thr Ala Thr Cys Gly Thr Ala Thr Gly Ala Thr Gly 50 55 60

Cys Ala Thr Thr Cys Thr Cys 65 70 <210> 58 <211> 71 <212> PRT <213> Artificial sequence <220><223> Synthetic peptide sequence <400>

Thr Cys Gly Ala Gly Ala Gly Ala Ala Thr Gly Cys Ala Thr Cys Ala 15 10 15

Thr Ala Cys Gly Ala Thr Ala Ala Gly Thr Ala Cys Gly Ala Ala Gly 20 25 30

Cys Cys Ala Thr Thr Cys Ala Cys Gly Ala Ala Gly Ala Gly Thr Thr 35 40 45

Gly Gly Ala Cys Cys Ala Thr Cys Ala Cys Gly Thr Gly Cys Cys Ala 50 55 60 -27- 1316961

Thr Ala Cys Cys Thr Thr Gly 65 70 <210> 59 <211> 77 <212> DNA <213>Artificial Sequence <220><223> Synthetic Peptide Sequence <220><221> CDS <222> (3)..(77) <223><400> 59 gt gca caa ggt atg gca cgt gat ggt cca act ctt cgt gaa tgg ctt 47 Ala Gin Gly Met Ala Arg Asp Gly Pro Thr Leu Arg Glu Trp Leu 15 10 15 cgt act tat cgt atg atg cat tct etc gag 77

Arg Thr Tyr Arg Met Met His Ser Leu Glu 20 25 <210> 60 <211> 25 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400> 60

Ala Gin Gly Met Ala Arg Asp Gly Pro Thr Leu Arg Glu Trp Leu Arg 15 10 15

Thr Tyr Arg Met Met His Ser Leu Glu 20 25 <210> 61 <211> 71 <212> PRT <213>Artificial Sequence<220><223> Synthetic peptide sequence-28 - 016732 1316961 <lt;;400> 61

Thr Gly Cys Ala Cys Ala Ala Gly Gly Ala Thr Gly Gly Ala Thr Gly 15 10 15

Cys Cys Ala Gly Ala Ala Gly Gly Ala Cys Cys Ala Ala Cys Ala Thr 20 25 30

Thr Ala Ala Ala Ala Cys Ala Ala Thr Gly Gly Cys Thr Thr Thr Thr 35 40 45

Thr Cys Ala Thr Gly Gly Thr Cys Gly Thr Gly Gly Thr Cys Ala Ala 50 55 60

Cys Ala Thr Thr Cys Thr Cys 65 70 <210> 62 <211> 71 <212> PRT <213> Artificial sequence <220><223> Synthetic peptide sequence <400>

Thr Cys Gly Ala Gly Ala Gly Ala Ala Thr Gly Thr Thr Gly Ala Cys 15 10 15

Cys Ala Cys Gly Ala Cys Cys Ala Thr Gly Ala Ala Ala Ala Ala Gly 20 25 30

Cys Cys Ala Thr Thr Gly Thr Thr Thr Thr Ala Ala Thr Gly Thr Thr 35 40 45

Gly Gly Thr Cys Cys Thr Thr Cys Thr Gly Gly Cys Ala Thr Cys Cys 50 55 60

Ala Thr Cys Cys Thr Thr Gly 65 70 <210> 63 <21i> 77 016733 29· 1316961 <212> DNA <213>Artificial Sequence <220><223> Synthetic Peptide Sequence <220><221> CDS <222> (3)..(77) <223><400> 63 gt gca caa gga tgg atg cca gaa gga cca aca tta aaa caa tgg ctt 47 Ala Gin Gly Trp Met Pro Glu Gly Pro Thr Leu Lys Gin Trp Leu 15 10 15 ttt cat ggt cgt ggt caa cat tct etc gag 77

"health sequence" <220> artificial sequence &lt

Ala Gin Gly Trp Met Pro Glu Gly Pro Thr Leu Lys Gin Trp Leu Phe 15 10 15

His Gly Arg Gly Gin His Ser Leu Glu 20 '25 <210> 65 <211> 71 <212> PRT .<213>Artificial Sequence <220><223> Synthetic Peptide Sequence <400> 65

Thr Gly Cys Ala Cys Ala Ala Gly Gly Ala Cys Ala Thr Ala Thr Thr 15 10 15

Cys Gly Thr Gly Ala Ala Gly Gly Thr Cys Cys Ala Ala Cys Ala Thr ΟΙΡΠ^Α -30- 1316961 20 25 30

Thr Ala Cys Gly Thr Cys Ala Ala Thr Gly Gly Cys Thr Thr Gly Thr 35 40 45

Thr Gly Cys Thr Cys Thr Thr Cys Gly Thr Ala Thr Gly Gly Thr Thr 50 55 60

Cys Ala Thr Thr Cys Thr Cys 65 70 <210> 66 <211> 71 <212> PRT <213> Artificial Sequence <220> -<223> Synthetic Peptide Sequence <400>

Thr Cys Gly Ala Gly Ala Gly Ala Ala Thr Gly Ala Ala Cys Cys Ala 15 10 15

Thr Ala Cys Gly Ala Ala Gly Ala Gly Cys Ala Ala Cys Ala Ala Gly 20 25 30

Cys Cys Ala Thr Thr Gly Ala Cys Gly Thr Ala Ala Thr Gly Thr Thr 35 40 45

Gly Gly Ala Cys Cys Thr Thr Cys Ala Cys Gly Ala Ala Thr Ala Thr 50 55 60

Gly Thr Cys Cys Thr Thr Gly 65 70 <210> 67 <211> 77 <212> DNA <213>Artificial Sequence <220><223> Synthetic peptide sequence <220> 016735 -31 - 1316961 <221> CDS <222>(3)„(77) <223><400> 67 gt gca caa gga cat att cgt gaa ggt cca aca tta cgt caa tgg ctt 47 Ala Gin Gly His lie Arg Glu Gly Pro Thr Leu Arg Gin Trp Leu 15 10 15 gtt get ctt cgt atg gtt cat tet etc gag 77

Val Ala Leu Arg Met Val His Ser Leu Glu 20 25 <210> 68 <211> 25 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence. <400> 68

Ala Gin Gly His lie Arg Glu Gly Pro Thr Leu Arg Gin Trp Leu Val 15 10 15

Ala Leu Arg Met Val His Ser Leu Glu 20 25 <210> 69 <211> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400> 69

Thr Gly Cys Ala Cys Ala Ala Gly Gly Thr Cys Ala Ala Thr Thr Ala 15 10 15

Gly Gly Ala Cys Ala Thr Gly Gly Thr Cys Cys Ala Ala Cys Thr Cys 20 25 30

Thr Thr Cys Gly Thr Cys Ala Ala Thr Gly Gly Cys Thr Thr Thr Cys 35 40 45 -32· 016736 1316961

Thr Thr Gly Gly Thr Ala Thr Cys Gly Thr Gly Gly Thr Ala Thr Gly 50 55 60

70, <210> 70 &lt

Thr Cys Gly Ala Gly Ala Gly Ala Ala Thr Gly Cys Ala Thr Ala Cys 15 10 15

Cys Ala Cys Gly Ala Thr Ala Cys Cys Ala Ala Gly Ala Ala Ala Gly 20 25 30

Cys Cys Ala Thr Thr Gly Ala Cys Gly Ala Ala Gly Ala GLy Thr Thr 35 40 45

Gly Gly Ala Cys Cys Ala Thr Gly Thr Cys Cys Thr Ala Ala Thr Thr 50 55 60

Gly Ala Cys Cys Thr Thr Gly 65 70 <210> 71 <211> 77 <212> DNA <213>Artificial Sequence <220><223> Synthetic Peptide Sequence <220><221> CDS <222> (3)..(77) <223><400> 71 gt gca caa ggt caa tta gga cat ggt cca act ctt cgt caa tgg ctt 47 Ala Gin Gly Gin Leu Gly His Gly Pro Thr Leu Axg Gin Trp Leu -33- 016737 1316961 15 10 15 tct tgg tat cgt ggt atg cat tct etc gag 71

Ser Trp Tyr Arg Gly Met His Ser Leu Glu 20 25 <210> 72 <211> 25 <212>PRT <2I3> Artificial Sequence <220><223> Synthetic Peptide Sequence <400>

Ala Gin Gly Gin Leu Gly His Gly Pro Thr Leu Arg Gin Trp Leu Ser 1 5 10 15

Trp Tyr Arg Gly Met His Ser Leu Glu 20 25

<210> 73 <211> 71 <212> PRT <213> Artificial sequence <220><223> Synthetic peptide sequence <400> 73

Thr Gly Cys Ala Cys Ala Ala Gly Gly Ala Gly Ala Ala Thr Thr Ala 15 10 15

Cys Giy Thr Cys Ala Ala Gly Gly Ala Cys Cys Ala Ala Cys Thr Cys 20 25 30

Thr Thr Cys Ala Thr Gly Ala Ala Thr Gly Gly Cys Thr Thr Cys Ala 35 40 45

Ala Cys Ala Thr Thr Thr Ala Gly Cys Ala Ala Gly Cys Ala Ala Ala 50 55 60

Cys Ala Thr Thr Cys Thr Cys 65 70 01G738 -34- 1316961 <210> 74 <211> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>; 74

Thr Cys Gly Ala Gly Ala Gly Ala Ala Thr Gly Thr Thr Thr Gly Cys 15 10 15

Thr Thr Gly Cys Thr Ala Ala Ala Thr Gly Thr Thr Gly Ala Ala Gly 20 25 30

Cys Cys Ala Thr Thr Cys Ala Thr Gly Ala Ala Gly Ala Gly Thr Thr 35 40 - 45

Gly Gly Thr Cys Cys Thr Thr Gly Ala Cys Gly Thr Ala Ala Thr Thr 50 55 60

Cys Thr Cys Cys Thr Thr Gly 65 70 <210> 75 <211> 77 <212> DNA <213>Artificial Sequence <220><223> Synthetic Peptide Sequence <220><221> CDS <222> (3),.(77) <223><400> 75 gt gca caa gga gaa tta cgt caa gga cca act ctt cat gaa tgg ctt 47 Ala Gin Gly Glu Leu Arg Gin Gly Pro Thr Leu His Glu Trp Leu 15 10 15 caa cat tta gca age aaa cat tet etc gag Gin His Leu Ala Ser Lys His Ser Leu Glu 20 25 -35 · 016739 77 1316961 <210> 76 <211> 25 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Ala Gin Gly Glu Leu Axg Gin Gly Pro Thr Leu His Glu Trp Leu Gin 15 10 15

His Leu Ala Ser Lys His Ser Leu Glu 20 25 <210> 77 <211> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400> 77 '

Thr Gly Cys Ala Cys Ala Ala Gly Gly Ala Gly Thr Ala Gly Gly Thr 15 10 15

Ala Thr Thr Gly Ala Ala Gly Gly Thr Cys Cys Ala Ala Cys Ala Thr 20 25 30

Thr Ala Cys Gly Thr Cys Ala Ala Thr Gly Gly Thr Thr Ala Gly Cys 35 40 45

Thr Cys Ala Ala Cys Gly Thr Cys Thr Thr Ala Ala Thr Cys Cys Ala 50 55 60

Cys Ala Thr Thr Cys Thr Cys 65 70 <210> 78 <211> 71 <212> PRT <213>Artificial Sequence<220> 016740 36-1316961 <223>Synthetic peptide sequence <400> 78

Thr Cys Gly Ala Gly Ala Gly Ala Ala Thr Gly Thr Gly Gly Ala Thr 15 10 15

Thr Ala Ala Gly Ala Cys Gly Thr Thr Gly Ala Gly Cys Thr Ala Ala 20 25 30

Cys Cys Ala Thr Thr Gly Ala Cys Gly Thr Ala Ala Thr Gly Thr Thr 35 40 45

Gly Gly Ala Cys Cys Thr Thr Cys Ala Ala Thr Ala Cys Cys Thr Ala 50 55 60

Cys Thr Cys Cys Thr Thi* Gly 65 70 <210> 79 <211> 77 <212> DNA <213>Artificial sequence <220><223> Synthetic peptide sequence <220> 〉CDS <222>(3)..(77) <223><400> 79 gt gca caa gga gta ggt att gaa ggt cca aca tta cgt caa tgg tta 47 Ala Gin Gly Val Gly lie Glu Gly Pro Thr Leu Arg Gin Trp Leu 15 10 15 get caa cgt ett aat cca cat tet etc gag 77

Ala Gin Arg Leu Asn Pro His Ser Leu Glu 20 25 <2i0> 80 <211> 25 <212> PRT <213>Artificial Sequence<220><223> Synthetic peptide sequence 37-016741 1316961 <;400> 80

Ala Gin Gly Val Gly lie Glu Gly Pro Thr Leu Arg Gin Trp Leu Ala 15 10 15

Gin Arg Leu Asn Pro His Ser Leu Glu 20 25 <210> 81 <211> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Thr Gly Cys Ala Cys Ala Ala Gly Gly Ala Thr Gly Gly Thr Cys Ala 15 10 15

Cys Gly Thr Gly Ala Thr Gly Gly Thr Cys Cys Ala Ala Cys Ala Cys 20 25 30

Thr Thr Cys Gly Thr Gly Ala Ala Thr Gly Gly Cys Thr Thr Gly Cys 35 40 45

Thr Thr Gly Gly Cys Gly Thr Gly Cys Thr Gly Thr Thr Gly Gly Ala 50 55 60

Cys Ala Thr Ala Gly Thr Cys 65 70 <210> 82 <211> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400> 82

Thr Cys Gly Ala Gly Ala Cys Thr Ala Thr Gly Thr Cys Cys Ala Ala 15 10 15 -38- 016742 1316961

Cys Ala Gly Cys Ala Cys Gly Cys Cys Ala Ala Gly Cys Ala Ala Gly 20 25 30

Cys Cys Ala Thr Thr Cys Ala Cys Gly Ala Ala Gly Thr Gly Thr Thr 35 40 45

Gly Gly Ala Cys Cys Ala Thr Cys Ala Cys Gly Thr Gly Ala Cys Cys 50 55 60

Ala Thr Cys Cys Thr Thr Gly 65 70 <210> S3 <211> 77 <212> DNA <213>Artificial Sequence <220><223> Synthetic Peptide Sequence <220><221> CDS <222> (3)..(77) <223><400> 83 gt gca caa gga tgg tea cgt gat ggt cca aca ett cgt gaa tgg ett 47 Ala Gin Gly Trp Ser Arg Asp Gly Pro Thr Leu Arg Glu Trp Leu 15 10 15 get tgg cgt get gtt gga cat agt etc gag 77

Ala Trp Arg Ala Val Gly His Ser Leu Glu 20 25 <210> 84 <211> 25 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Ala Gin Gly Trp Ser Arg Asp Gly Pro Thr Leu Arg Glu Trp Leu Ala 15 10 15 -39- 016743 1316961

Trp Arg Ala Val Gly His Ser Leu Glu 20 25 <210> 85 <2il> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Thr Gly Cys Ala Cys Ala Ala Gly Gly Ala Gly Cys Ala Gly Thr Thr 15 10 15

Cys Cys Ala Cys Ala Ala Gly Gly Ala Cys Cys Ala Ala Cys Thr Cys 20 25 30

Thr Thr Ala Ala Ala Cys Ala Gly Thr Gly Gly Thr Thr Ala Thr Thr 35 40 45

Ala Thr Gly Gly Cys Gly Thr Cys Gly Thr Thr Gly Thr Gly Cys Ala 50 55 60

Cys Ala Thr Thr Cys Thr Cys 65 70 <210> 86 <211> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Thr Cys Gly Ala Gly Ala Gly Ala Ala Thr Gly Thr Gly Cys Ala Cys 15 10 15

Ala Aia Cys Gly Ala Cys Gly Cys Cys Ala Thr Ala Ala Thr Ala Ala 20 25 30

Cys Cys Ala Cys Thr Gly Thr Thr Thr Ala Ala Gly Ala Gly Thr Thr 35 40 45 016744 -40- 1316961

Gly Gly Thr Cys Cys Thi· Thr Gly Thr Gly Gly Ala Ala Cys Thr Gly 50 55 60

Cys Thr Cys Cys Thr Thr Gly 65 70 <210> 87 <211> 77 <212> DNA <213>Artificial sequence <220><223> Synthetic peptide sequence <220>

<221> CDS <222> (3)..(77) ' <223><400> 87 gt gca caa gga gca gtt cca caa gga cca act ctt aaa cag tgg tta 47 Ala Gin Gly Ala Val Pro Gin Gly Pro Thr Leu Lys Gin Trp Leu 15 10 15 tta tgg cgt cgt tgt gca cat tct etc gag 77

Leu Trp Arg Arg Cys Ala His Ser Leu Glu 20 25 <210> 88 <211> 25 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Ala Gin Gly Ala Val Pro Gin Gly Pro Thr Leu Lys Gin Trp Leu Leu 15 10 15

Trp Arg Arg Cys Ala His Ser Leu Glu 20 25

<210> 89 <2ll> 71 <212> PRT -41 - 016745 1316961 <213>Artificial sequence <220><223> Synthetic surname peptide sequence <400>

Thr Gly Cys Ala Cys Ala Ala Gly Gly Thr Cys Gly Thr Ala Thr Thr 15 10 15

Cys Gly Thr Gly Ala Ala Gly Gly Thr Cys Cys Ala Ala Cys Thr Cys 20 25 30

Thr Thr Ala Ala Ala Gly Ala Ala Thr Gly Gly Cys Thr Thr Gly Cys 35 40 45

Thr Cys Ala Ala Cys Gly Thr Cys Gly Thr Gly Gly Thr Thr Thr Thr 50 55 - 60

Cys Ala Thr Ala Gly Thr Cys 65 70 <210> 90 <211> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Thr Cys Gly Ala Gly Ala Cys Thr Ala Thr Gly Ala Ala Ala Ala Cys 15 10 15

Cys Ala Cys Gly Ala Cys Gly Thr Thr Gly Ala Gly Cys Ala Ala Gly 20 25 30

Cys Cys Ala Thr Thr Cys Thr Thr Thr Ala Ala Gly Ala Gly Thr Thr 35 40 45

Gly Gly Ala Cys Cys Thr Thr Cys Ala Cys Gly Ala Ala Thr Ala Cys 50 55 60

Gly Ala Cys Cys Thr Thr Gly 016746 42- 1316961 65 70 <210> 91 <211> 77 <212> DNA <213>Artificial sequence <220><223> Synthetic peptide sequence <220><221> CDS <222>(3)..(77) <223><400> 91 gt gca caa ggt cgt att cgt gaa ggt cca act ctt aaa gaa tgg ctt 47 Ala Gin Gly Arg lie Arg Glu Gly Pro Thr Leu Lys Glu Trp Leu 15 10 15 get caa cgt cgt ggt ttt cat agt etc gag 77

Ala Gin Arg Arg Gly Phe His Ser Leu Glu 20 25 <210> 92 <211> 25 <212> PRT <213>Artificial Sequence<220><223> Synthetic Victory Too Sequence<400> 92

Ala Gin Gly Arg lie Arg Glu Gly Pro Thr Leu Lys Glu Trp Leu Ala 15 10 15

Gin Arg Arg Gly Phe His Ser Leu Glu 20 25 <210> 93 <211> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400> 93 - 43 - 016747 1316961

Thr Gly Cys Ala Cys Ala Ala Gly Gly Thr Cys Gly Thr Thr Thr Cys 15 10 15

Gly Cys Thr Gly Ala Ala Gly Gly Thr Cys Cys Ala Ala Cys Ala Cys 20 25 30

Thr Thr Cys Gly Thr Gly Ala Ala Thr Gly Gly Thr Thr Ala Gly Ala 35 40 45

Ala Cys Ala Ala Cys Gly Thr Ala Ala Ala Cys Thr Thr Gly Thr Thr 50 55 60

Cys Ala Thr Ala Gly Thr Cys 65 70 <210> 94 <211> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400> 94

Thr Cys Gly Ala Gly Ala Cys Thr Ala Thr Gly Ala Ala Cys Ala Ala 15 10 15

Gly Thr Thr Thr Ala Cys Gly Thr Thr Gly Thr Thr Cys Thr Ala Ala 20 25 30

Cys Cys Ala Thr Thr Cys Ala Cys Gly Ala Ala Gly Thr Gly Thr Thr 35 40 45

Gly Gly Ala Cys Cys Thr Thr Cys Ala Gly Cys Gly Ala Ala Ala Cys 50 55 60

Gly Ala Cys Cys Thr Thr Gly 65 70 <210> 95 <211> 77 <212> DNA <213>Artificial sequence 016748 -44 - 1316961 <220><223> Synthetic peptide sequence <220>;<221> CDS <222> (3)..(77) <223><400> 95 gt gca caa ggt cgt ttc get gaa ggt cca aca ett cgt gaa tgg tta 47 Ala Gin Gly Arg Phe Ala Glu Gly Pro Thr Leu Arg Glu Trp Leu 15 10 15 gaa caa cgt aaa ett gtt cat agt etc gag 77

Glu Gin Arg Lys Leu Val His Ser Leu Glu 20 25 <210> 96 <211> 25 * <212> PRT <213>Artificial Sequence <220><223> Synthetic Peptide Sequence <400> 96

Ala Gin Gly Arg Phe Ala Glu Gly Pro Thr Leu Arg Glu Tip Leu Glu 15 10 15

Gin Arg Lys Leu Val His Ser Leu Glu 20 25 <210> 97 <211> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Thr Gly Cys Ala Cys Ala Ala Gly Gly Thr Gly Ala Thr Cys Gly Thr 15 10 15

Thr Thr Cys Cys Ala Ala Gly Gly Thr Cys Cys Ala Ala Cys Thr Cys 20 25 30 016749 -45 - 1316961

Thr Thr Cys Gly Thr Gly Ala Ala Thr Gly Gly Cys Thr Thr Gly Cys 35 40 45

Thr Gly Cys Ala Ala Thr Cys Cys Gly Thr Ala Gly Cys Gly Thr Ala 50 55 60

Cys Ala Tlu Ala Gly Thr Cys 65 70 <210> 98 <211> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400> 98

Thr Cys Gly Ala Gly Ala Cys Thr Ala Thr Gly Thr Ala Cys Gly Cys 15 10 15

Thr Ala Cys Gly Gly Ala Thr Thr Gly Cys Ala Gly Cys Ala Ala Gly 20 25 30

Cys Cys Ala Thr Thr Cys Ala Cys Gly Ala Ala Gly Ala Gly Thr Thr 35 40 45

Gly Gly Ala Cys Cys Thr Thr Gly Gly Ala Ala Ala Cys Gly Ala Thr 50 55 60

Cys Ala Cys Cys Thr Thr Gly 65 70 <210> 99 <211> 77 <212> DNA <213>Artificial Sequence <220><223> Synthetic Peptide Sequence <220><221> CDS <222>(3)., (77) 016750 -46- 1316961 <223><400> 99 gt gca caa ggt gat cgt ttc caa ggt cca act ctt cgt gaa tgg ctt 47 Ala Gin Gly Asp Arg Phe Gin Gly Pro Thr Leu Arg Glu Trp Leu 15 10 15 get gca ate cgt age gta cat agt etc gag 77

Ala Ala lie Arg Ser Val His Ser Leu Glu 20 25 <210> 100 <211> 25 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400> -

Ala Gin Gly Asp Arg Phe Gin Gly Pro Thr Leu Arg Glu Trp Leu Ala 15 10 15

Ala lie Arg Ser Val His Ser Leu Glu 20 25 <210> 101 <211> 71 <212> PRT <213>Artificial sequence <220><223>Synthesis peptide sequence <400>

Thr Gly Cys Ala Cys Ala Ala Gly Gly Thr Gly Cys Thr Gly Gly Thr 15 10 15

Cys Gly Thr Gly Ala Ala Gly Gly Thr Cys Cys Ala Ala Cys Thr Cys 20 25 30

Thr Ala Cys Gly Thr Gly Ala Ala Thr Gly Gly Cys Thr Thr Ala Ala 35 40 45

Thr Ala Thr Gly Cys Gly Thr Gly Thr Thr Thr Gly Gly Cys Ala Ala 50 55 60 -47- 016751 1316961

Cys Ala Thr Thr Cys Thr Cys 65 70 <210> 102 <211> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400> 102

Thr Cys Gly Ala Gly Ala Gly Ala Ala Thr Gly Thr Thr Gly Cys Cys 15 10 15

Ala Ala Ala Cys Ala Cys Gly Cys Ala Thr Ala Thr Thr Ala Ala Gly 20 25 30

Cys Cys Ala Thr Thr Cys Ala Cys Gly Thr Ala Gly Ala Gly Thr Thr 35 40 45

Gly Gly Ala Cys Cys Thr Thr Cys Ala Cys Gly Ala Cys Cys Ala Gly 50 55 60

Cys Ala Cys Cys Thr Thr Gly 65 70 <210> 103 <211> 77 <212> DNA <213>Artificial sequence <220><223> Synthetic peptide sequence <220><221> CDS <222> 0)..(77) <223><400> 103 gt gca caa ggt get ggt cgt gaa ggt cca act eta cgt gaa tgg ett 47 Ala Gin Gly Ala Gly Arg Glu Gly Pro Thr Leu Arg Glu Trp Leu 15 10 15 -48- 016752 1316961 aat atg cgt gtt tgg caa cat tct etc gag 77

Asn Met Arg Val Trp Gin His Ser Leu Glu 20 25 <210> 104 <211> 25 <212> PRT <213>Artificial sequence <220><223> Synthetic skin sequence <400>

Ala Gin Gly Ala Gly Arg Glu Gly Pro Thr Leu Arg Glu Trp Leu Asn 1 5 10 15

Met Arg Val Trp Gin His Ser Leu Glu 20 25 - <210> 105 <211> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Thr Gly Cys Ala Cys Ala Ala Gly Gly Ala Gly Cys Thr Thr Thr Ala 15 10 15

Cys Ala Ala Gly Ala Ala Gly Gly Ala Cys Cys Ala Ala Cys Ala Thr 20 25 30

Thr Ala Cys Gly Thr Cys Ala Ala Thr Gly Gly Thr Thr Ala Gly Gly 35 40 45

Ala Thr Gly Gly Gly Gly Thr Cys Ala Ala Thr Gly Gly Gly Gly Ala 50 55 60

Cys Ala Cys Thr Cys Thr Cys 65 70 <210> 106 016753 49-1316961 <211> 71 <212> PRT <213>Artificial Sequence<220><223> Synthetic peptide sequence <400> 106

Thr Cys Gly Ala Gly Ala Gly Ala Gly Thr Gly Thr Cys Cys Cys Cys 15 10 15

Ala Thr Thr Gly Ala Cys Cys Cys Cys Ala Thr Cys Cys Thr Ala Ala 20 25 30

Cys Cys Ala Thr Thr Gly Ala Cys Gly Thr Ala Ala Thr Gly Thr Thr 35 40 45

Gly Gly Thr Cys Cys Tta Thr Cys Thr Thr Gly Thr Ala Ala Ala Gly 50 55 60

Cys Thr Cys Cys Thr Thr Gly 65 70 <210> 107 <211> 77 <212> DNA <213>Artificial Sequence <220><223> Synthetic Peptide Sequence <220><221> CDS <222>(3)..(77) <223><400> 107 gt gca caa gga get tta caa £aa gga cca aca tta cgt caa tgg tta 47 Ala Gin Gly Ala Leu Gin Glu Gly Pro Thr Leu Arg Gin Trp Leu 1 5 10 15 gga tgg ggt caa tgg gga cac tet etc gag Gly Trp Gly Gin Trp Gly His Ser Leu Glu 20 25 <210> 108 <211> 25

广,6W -50- 77 1316961 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Ala Gin Gly Ala Leu Gin Glu Gly Pro Thr Leu Arg Gin Trp Leu Gly 15 10 15

Trp Gly Gin Trp Gly His Ser Leu Glu 20 25 <210> 109 <211> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Thr Gly Cys Ala Cys Ala Ala Gly Gly Ala Thr Ala Cys Thr Gly Thr 15 10 15

Gly Ala Thr Gly Ala Ala Gly Gly Thr Cys Cys Ala Ala Cys Thr Cys 20 25 30

Thr Thr Ala Ala Ala Cys Ala Ala Thr Gly Gly Thr Thr Ala Gly Thr 35 40 45

Ala Thr Gly Thr Cys Thr Thr Gly Gly Thr Thr Thr Ala Cys Ala Ala 50 55 60

Cys Ala Thr Ala Gly Thr Cys 65 70 <210> 110 <211> 71 <212> PRT <213>Artificial Sequence<220><223> Synthetic peptide sequence -51 - 016755 1316961 <400&gt ; 110

Thr Cys Gly Ala Gly Ala Cys Thr Ala Thr Gly Thr Thr Gly Thr Ala 15 10 15

Ala Ala Cys Cys Ala Ala Gly Ala Cys Ala Thr Ala Cys Thr Ala Ala 20 25 30

Cys Cys Ala Thr Thr Gly Thr Thr Thr Ala Ala Gly Ala Gly Thr Thr 35 40 45

Gly Gly Ala Cys Cys Thr Thr Cys Ala Thr Cys Ala Cys Ala Gly Thr 50 55 60

Ala Thr Cys Cys Thr Thr Gly 65 70 <210> 111 <211> 77 <212> DNA <213>Artificial Sequence <220><223> Synthetic Peptide Sequence <220><221> CDS <222> (3)..(77) <223><400> 111 gt gca caa gga tac tgt gat gaa ggt cca act ctt aaa caa tgg tta 47 Ala Gin Gly Tyr Cys Asp Glu Gly Pro Thr Leu Lys Gin Trp Leu 15 10 15 gta tgt ctt ggt tta caa cat agt etc gag 77

Val Cys Leu Gly Leu Gin His Ser Leu Glu 20 25 <210> 112 <211> 25 <212> PRT < 213 > Artificial Sequence <220><223> Synthetic Peptide Sequence <400> -52· 016756 1316961

Ala Gin Gly Tyr Cys Asp Glu Gly Pro Thr Leu Lys Gin Trp Leu Val 15 10 15

Cys Leu Gly Leu Gin His Ser Leu Glu 20 25 <210> 113 <2U> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Thr Gly Cys Ala Cys Ala Ala Gly Gly Ala Thr Gly Thr Ala Gly Thr 15 10 15

Thr Cys Ala Gly Gly Ala Gly Gly Thr Cys Cys Ala Ala Cys Thr Thr 20 25 30

Thr Ala Cys Gly Thr Gly Ala Ala Thr Gly Gly Thr Thr Ala Cys Ala 35 40 45

Ala Thr Gly Thr Cys Gly Thr Cys Gly Thr Ala Thr Gly Cys Ala Ala 50 55 60

Cys Ala Thr Thr Cys Thr Cys 65 70 <210> 114 <211> 71 <212>PRT <213> Artificial sequence <220><223> Synthetic peptide sequence <400>

Thr Cys Gly Ala Gly Ala Gly Ala Ala Thr Gly Thr Thr Gly Cys Ala 15 10 15

Thr Ala Cys Gly Ala Cys Gly Ala Cys Ala Thr Thr Gly Thr Ala Ala -53- 016757 1316961 20 25 30

Cys Cys Ala Thr Thr Cys Ala Cys Gly Thr Ala Ala Ala Gly Thr Thr 35 40 45

Gly Gly Ala Cys Cys Thr Cys Cys Thr Gly Aia Ala Cys Thr Ala Cys 50 55 60

Ala Thr Cys Cys Thr Thr Gly 65 70 <210> 115 <211> 77 <212> DNA <213>Artificial Sequence <220> - <223> Synthetic Peptide Sequence <220><221> CDS <222> (3)..(77) <223><400> 115 gt gca caa gga tgt agt tea gga ggt cca act tta cgt gaa tgg tta 47 Ala Gin Gly Cys Ser Ser Gly Gly Pro Thr Leu Arg Glu Trp Leu 15 10 15 caa tgt cgt cgt atg caa cat tet etc gag 77

Gin Cys Arg Arg Met Gin His Ser Leu Glu 20 25 <210> 116 <211> 25 <212> PRT <213> Artificial sequence <220><223> Synthetic peptide sequence <400>

Ala Gin Gly Cys Ser Ser Gly Gly Pro Thr Leu Arg Glu Trp Leu Gin 15 10 15

Cys Arg Arg Met Gin His Ser Leu Glu 20 25 -54- 016758 1316961 <210> 117 <211> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <223>;400> 117

Thr Gly Cys Ala Cys Ala Ala Gly Gly Ala Thr Gly Thr Thr Cys Ala 15 10 15

Thr Gly Gly Gly Gly Thr Gly Gly Thr Cys Cys Ala Ala Cys Thr Cys 20 25 30

Thr Thr Ala Ala Ala Cys Ala Ala Thr Gly Gly Thr Thr Ala Cys Ala 35 40 " 45

Ala Thr Gly Thr Gly Thr Thr Cys Gly Thr Gly Cys Thr Ala Ala Ala 50 55 60

Cys Ala Thr Thr Cys Thr Cys 65 70 <210> 118 <211> 71 <212> PRT <213> Artificial sequence <220><223> Synthetic peptide sequence _ <400>

Thr Cys Gly Ala Gly Ala Gly Ala Ala Thr Gly Thr Thr Thr Ala Gly 15 10 15

Cys Ala Cys Gly Ala Ala Cys Ala Cys Ala Thr Thr Gly Thr Ala Ala 20 25 30

Cys Cys Ala Thr Thr Gly Thr Thr Thr Ala Ala Gly Ala Gly Thr Thr 35 40 45 016759 -55 - 1316961

Gly Gly Ala Cys Cys Ala Cys Cys Cys Cys Ala Thr Gly Ala Ala Cys 50 55 60

Ala Thr Cys Cys Thr Thr Gly 65 70 <210> 119 <211> 77 <212> DNA <213>Artificial Sequence <220><223> Synthetic Peptide Sequence <220><221> CDS <222> (3)..(77) <223><400> 119 gt gca caa gga tgt tea tgg ggt ggt cca act ett aaa caa tgg tta 47 Ala Gin Gly Cys Ser Trp Gly Gly Pro Thr Leu Lys Gin Trp Leu 15 10 15 caa tgt gtt cgt get aaa cat tet etc gag 77

Gin Cys Val Arg Ala Lys His Ser Leu Glu 20 25 <210> 120 <211> 25 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Ala Gin Gly Cys Ser Trp Gly Gly Pro Thr Leu Lys Gin Trp Leu Gin 15 10 15

Cys Val Arg Ala Lys His Ser Leu Glu 20 25 <210> 121 <211> 71 <212> PRT <213> Artificial sequence 56-016760 1316961 <220><223> Synthetic peptide sequence< 400> 121

Thr Gly Cys Ala Cys Ala Ala Gly Gly Ala Thr Gly Thr Cys Ala Ala 15 10 15

Thr Thr Ala Gly Gly Thr Gly Gly Thr Cys Cys Gly Ala Cys Thr Cys 20 25 30

Thr Thr Cys Gly Thr Gly Ala Ala Thr Gly Gly Cys Thr Thr Gly Cys 35 40 45

Thr Thr Gly Thr Cys Gly Thr Cys Thr Thr Gly Gly Thr Gly Cys Thr 50 55 60

Cys Ala Thr Thr Cys Ala Cys 65 70 <210> 122 <211> 71 <212>PRT <213>Artificial sequence <220><223>Synthesis peptide sequence <400>

Thr Cys Gly Ala Gly Thr Gly Ala Ala Thr Gly Ala Gly Cys Ala Cys 15 10 15

Cys Ala Ala Gly Ala Cys Gly Ala Cys Ala Ala Gly Cys Ala Ala Gly 20 25 30

Cys Cys Ala Thr Thr Cys Ala Cys Gly Ala Ala Gly Ala Gly Thr Cys 35 40 45

Gly Gly Ala Cys Cys Ala Cys Cys Thr Ala Ala Thr Thr Gly Ala Cys 50 55 60

Ala Thr Cys Cys Thr Thr Gly 65 70 016761 -57- 1316961 <210> 123 <211> 77 <212> DNA <2}3>Artificial Sequence<220><223> Synthetic peptide sequence<;220><221> CDS <222> (3)..(77) <223><400> 123 gt gca caa gga tgt caa tta ggt ggt ccg act ctt cgt gaa tgg ctt 47 Ala Gin Gly Cys Gin Leu Gly Gly Pro Thr Leu Arg Glu Trp Leu 15 10 15 get tgt cgt ctt ggt get cat tea etc gag 77

Ala Cys Arg Leu Gly Ala His Ser Leu Glu 20 25 <210> 124 <211> 25 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Ala Gin Gly Cys Gin Leu Gly Gly Pro Thr Leu Arg Glu Trp Leu Ala 1 5 10 15

Cys Arg Leu Gly Ala His Ser Leu Glu 20 25 <210> 125 <211> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400> 125

Thr Gly Cys Ala Cys Ala Ala Gly Gly Ala Thr Gly Thr Thr Gly Gly 15 10 15 016762 • 58- 1316961

Gly Ala Ala Gly Gly Tl\r Gly Gly Thr Cys Cys Thr Ala Cys Ala Cys 20 25 30

Thr Thr Ala Ala Ala Gly Ala Ala Thr Gly Gly Cys Thr Thr Cys Ala 35 40 45

Ala Thr Gly Thr Cys Thr Thr Gly Thr Ala Gly Ala Ala Cys Gly Thr 50 55 60

Cys Ala Thr Thr Cys Ala Cys 65 70 <210> 126 <211> 71 <212> PRT . <213> Artificial sequence <220><223> Synthetic peptide sequence <400>

Thr Cys Gly Ala Gly Thr Gly Ala Ala Thr Gly Ala Cys Gly Thr Thr 15 10 15

Cys Thr Ala Cys Ala Ala Gly Ala Cys Ala Thr Thr Gly Ala Ala Gly 20 25 30

Cys Cys Ala Thr Thr Cys Thr Thr Thr Ala Ala Gly Thr Gly Thr Ala 35 40 45

Gly Gly Ala Cys Cys Ala Cys Cys Thr Thr Cys Cys Cys Ala Ala Cys 50 55 60

Ala Thr Cys Cys Thr Thr Gly 65 70 <210> 127 <211> 77 <212> DNA <213>Artificial Sequence<220> 016763 59-1316961 <223> Synthetic peptide sequence <220><221> CDS <222> (3)., (77) <223><400> 127 gt gca caa gga tgt tgg gaa ggt ggt cct aca ctt aaa gaa tgg ctt 47 Ala Gin Gly Cys Trp Glu Gly Gly Pro Thr Leu Lys Glu Trp Leu 15 10 15 caa tgt ctt gta gaa cgt cat tea etc gag 77

Gin Cys Leu Val Glu Arg His Ser Leu Glu 20 25

<210> 128 <211> 25 <212> PRT <213> Artificial sequence - <220><223> Synthetic peptide sequence <400>

Ala Gin Gly Cys Trp Glu Gly Gly Pro Thr Leu Lys Glu Trp Leu Gin 1 5 10 15

Cys Leu Val Glu Axg His Ser Leu Glu 20 25 <210> 129 <211> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Thr Gly Cys Ala Cys Ala Ala Gly Gly Thr Thr Gly Thr Cys Gly Tlor 15 10 15

Gly Gly Thr Gly Gly Thr Gly Gly Thr Cys Cys Ala Ala Cys Thr Cys 20 25 30

Thr Thr Cys Ala Thr Cys Ala Ala Thr Gly Gly Cys Thr Thr Thr Cys 016765 1316961 35 40 45

Thr Thr Gly Thr Thr Thr Thr Cys Gly Thr Thr Gly Gly Cys Ala Ala 50 55 60

Cys Ala Thr Thr Cys Ala Cys 65 70 <210> 130 <211> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Thr Cys Gly Ala Gly Thr Gly Ala Ala Thr Gly Thr Thr Gly Cys Cys 15 10 15

Ala Ala Cys Gly Ala Ala Ala Ala Cys Ala Ala Gly Ala Ala Ala Gly 20 25 30

Cys Cys Ala Thr Thr Gly Ala Thr Gly Ala Ala Gly Ala Gly Thr Thr 35 40 45

Gly Gly Ala Cys Cys Ala Cys Cys Ala Cys Cys Ala Cys Gly Ala Cys 50 55 60

Ala Ala Cys Cys Thr Thr Gly 65 70 <210> 131 <211> 77 <212> DNA <213>Artificial sequence <220><223> Synthetic peptide sequence <220><221> CDS <222> (3)..(77) <223> 016766 -61 - 1316961 <400> 131 gt gca caa ggt tgt cgt ggt ggt ggt cca act ctt cat caa tgg ctt 47 Ala Gin Gly Cys Arg Gly Gly Gly Pro Thr Leu His Gin Tip Leu 15 10 15 tct tgt ttt cgt tgg caa cat tea etc gag 77

Ser Cys Phe Arg Trp Gin His Ser Leu Glu 20 25 <210> 132 <211> 25 <212>PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400> 132

Ala Gin Gly Cys Arg Gly Gly Gly Pro Thr Leu His Gin Trp Leu Ser 15 10 15

Cys Phe Arg Trp Gin His Ser Leu Glu 20 25 <210> 133 <211> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Thr Gly Cys Ala Cys Ala Ala Gly Gly Ala Thr Gly Thr Cys Gly Thr 15 10 15

Gly Ala Thr Gly Gly Thr Gly Gly Thr Cys Cys Ala Ala Cys Thr Cys 20 25 30

Thr Thr Ala Gly Ala Cys Ala Ala Thr Gly Gly Cys Thr Thr Gly Cys 35 40 45

Thr Thr Gly Thr Cys Thr Thr Cys Ala Ala Cys Ala Ala Ala Ala Ala 50 55 60 -62- 016767 1316961

Cys Ala Thr Thr Cys Ala Cys 65 70 <210> 134 <211> 71 , <212> PRT < 213 > artificial sequence <220 < 223 > synthetic peptide sequence <400>

Thr Cys Gly Ala Gly Thr Gly Ala Ala Thr Gly Thr Thr Thr Thr Thr 1 5 10 15

Gly Thr Thr Gly Ala Ala Gly Ala Cys Ala Ala Gly Cys Ala Ala Gly 20 25 30

Cys Cys Ala Thr Thr Gly Thr Cys Thr Ala Ala Gly Ala Gly Thr Thr 35 40 45

Gly Gly Ala Cys Cys Ala Cys Cys Ala Thr Cys Ala Cys Gly Ala Cys 50 55 60

Ala Thr Cys Cys Thr Thr Gly 65 70 <210> 135 <211> 77 <212> DNA <213>Artificial Sequence <220><223> Synthetic B-peptide sequence <220><221> CDS <222> (3)..(77) <223><400> 135 gt gca caa gga tgt cgt gat ggt ggt cca act ctt aga caa tgg ctt 47 Ala Gin Gly Cys Arg Asp Gly Gly Pro Thr Leu Arg Gin Trp Leu 1 5 10 15 77 get tgt ctt caa caa aaa cat tea etc gag Ala Cys Leu Gin Gin Lys His Ser Leu Glu niR7i: 〇-63- 1316961 20 25 <210> 136 <211> 25 &lt ;212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Ala Gin Gly Cys Arg Asp Gly Gly Pro Thr Leu Arg Gin Trp Leu Ala 15 10 15

Cys Leu Gin Gin Lys His Ser Leu Glu 20 25 <210> 137 <211> 71 <212> PRT <213>Artificial Sequence <220><223> Synthetic Skin Sequence <400> 137

Thr Cys Gly Ala Gly Thr Gly Ala Ala Thr Gly Thr Thr Gly Ala Gly 15 10 15

Cys Ala Ala Gly Ala Cys Gly Cys Cys Ala Ala Ala Cys Ala Ala Gly 20 25 30

Cys Cys Ala Thr Thr Cys Thr Thr Thr Thr Ala Ala Ala Gly Thr Thr 35 40 45

Gly Gly Ala Cys Cys Ala Gly Ala Thr Cys Thr Thr Ala Ala Thr Thr 50 55 60

Cys Thr Cys Cys Thr Thr Gly 65 . 70

<210> 138 <211> 71 <212> PRT 016769 64· 1316961 <213> Artificial sequence <220><223> Synthetic peptide sequence <400>

Thr Gly Cys Ala Cys Ala Ala Gly Gly Ala Gly Ala Ala Thr Thr Ala 15 10 15

Ala Gly Ala Thr Cys Thr Gly Gly Thr Cys Cys Ala Ala Cys Thr Thr 20 25 30

Thr Ala Ala Ala Ala Gly Ala Ala Thr Gly Gly Cys Thr Thr Gly Thr 35 40 45

Thr Thr Gly Gly Cys Gly Thr Cys Thr Thr Gly Cys Thr Cys Ala Ala 50 55 - 60

Cys Ala Thr Thr Cys Ala Cys 65 70 <210> 139 <211> 77 <212> DNA <213>Artificial sequence <220><223> Synthetic peptide sequence <220><221> CDS <222>(3)..(77) <223><400> 139 gt gca caa gga gaa tta aga tct ggt cca act tta aaa gaa tgg ctt 47 Ala Gin Gly Glu Leu Arg Ser Gly Pro Thr Leu Lys Glu Trp Leu 15 10 15 gtt tgg cgt ctt get caa cat tea etc gag 77

Val Trp Arg Leu Ala Gin His Ser Leu Glu 20 25 <210> 140 <211> 25 <212> PRT <213>Artificial Sequence 016770 -65 - 1316961 <220><223> Synthetic peptide sequence <400> 140

Ala Gin Gly Glu Leu Arg Ser Gly Pro Thr Leu Lys Glu Trp Leu Val 15 10 15

Trp Arg Leu Ala Gin His Ser Leu Glu 20 25 <210> 141 <211> 71 <2i2> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence - <400>

Thr Gly Cys Ala Cys Ala Ala Gly Gly Ala Gly Gly Ala Thr Gly Thr 15 10 15

Ala Gly Ala Thr Cys Thr Gly Gly Thr Cys Cys Ala Ala Cys Ala Cys 20 25 30

Thr Thr Cys Gly Thr Gly Ala Ala Thr Gly Gly Thr Thr Ala Gly Cys 35 40 45

Thr Thr Gly Thr Ala Gly Ala Gly Ala Gly Gly Thr Thr Cys Ala Ala 50 55 60

Cys Ala Cys Thr Cys Thr Cys 65 70 <210> 142 <211> 71 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400> 142 016771 -66 - 1316961

Thr Cys Gly Ala Gly Ala Gly Ala Gly Thr Gly Thi Thr Gly Ala Ala 15 10 15

Cys Cys Thr Cys Thr Cys Thr Ala Cys Ala Ala Gly Cys Thr Ala Ala 20 25 30

Cys Cys Ala Thr Thr Cys Ala Cys Gly Ala Ala Gly Thr Gly Thr Thr 35 40 45

Gly Gly Ala Cys Cys Ala Gly Ala Thr Cys Thr Ala Cys Ala Thr Cys 50 55 60

Cys Thr Cys Cys Thr Thr Gly 65 70 <210> 143 <211> 77 <212> DNA <213>Artificial Sequence <220><223> Synthetic Peptide Sequence <220><221> CDS <222> (3)..(77) <223><400> 143 gt gca caa gga gga tgt aga tct ggt cca aca ctt cgt gaa tg£ tta 47 Ala Gin Gly Gly Cys Arg Ser Gly Pro Thr Leu Arg Glu Trp Leu 15 10 15 get tgt aga gag gtt caa cac tct etc gag 77

Ala Cys Arg Glu Val Gin His Ser Leu Glu 20 25 <210> 144 <211> 25 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Ala Gin Gly Gly Cys Arg Ser Gly Pro Thr Leu Arg Glu Trp Leu Ala 016772 -67- 1316961 15 10 15

Cys Arg Glu Val Gin His Ser Leu Glu 20 25 <210> 145 <211> 71 <212> PRT <213:-Artificial sequence <220><223> Synthetic peptide sequence <400>

Thr Gly Cys Ala Cys Ala Ala Gly Gly Thr Ala Cys Ala Thr Gly Cys 15 10 15

Gly Ala Ala Cys Ala Ala Gly Gly Ala Cys Cys Ala Ala Cys Thr Cys 20 25 30

Thr Ala Ala Gly Ala Cys Ala Ala Thr Gly Gly Cys Thr Ala Cys Thr 35 40 45

Ala Thr Gly Thr Ala Gly Ala Cys Ala Ala Giy Gly Ala Ala Gly Ala 50 55 60

Cys Ala Cys Thr Cys Ala Cys 65 70 <210> 146 <211> 71 <212> PRT < 213: artificial sequence <220><223> synthetic peptide sequence <400>

Thr Cys Gly Ala Gly Thr Gly Ala Gly Thr Gly Thr Cys Thr Thr Cys 15 10 15

Cys Thr Thr Gly Thr Cys Thr Ala Cys Ala Thr Ala Gly Thr Ala Gly 20 25 30 016773 -68 - 1316961

Cys Cys Ala Thr Thr Gly Thr Cys Thr Thr Ala Gly Ala Gly Thr Thr 35 40 45

Gly Gly Thr Cys Cys Thr Thr Gly Thr Thr Cys Gly Cys Ala Thr Gly 50 55 60

Thr Ala Cys Cys Thr Thr Gly 65 70 <210> 147 <211> 77 <212>DNA <213>Artificial Sequence <220><223> Synthetic Peptide Sequence <220><221> CDS <222> (3)..(77) <223><400> 147 gt gca caa ggt aca tgc gaa caa gga cca act eta aga caa tgg eta 47 Ala Gin Gly Thr Cys Glu Gin Gly Pro Thr Leu Arg Gin Trp Leu 15 10 15 eta tgt aga caa gga aga cac tea etc gag 77

Leu Cys Arg Gin Gly Arg His Ser Leu Glu 20 25 <210> 148' <211> 25 <212>PRT <213>Artificial Sequence<220><223> Synthetic Victory Sequence <40ϋ> 148

Ala Gin Gly Thr Cys Glu Gin Gly Pro Thr Leu Arg Gin Trp Leu Leu 15 10 15

Cys Arg Gin Gly Arg His Ser Leu Glu 20 25 016774 -69- 1316961 <210> 149 <211> 77 <212> DMA <213>Artificial Sequence <220><223> Synthetic Victory Sequence <223>;220><221> CDS <222> (3)..(77) <223><400> 149 gt gca cag ggt tgg tgt aag gag ggt cct act ctg cgt gag tgg ctg 47 Ala Gin Gly Trp Cys Lys Glu Gly Pro Thr Leu Arg Glu Trp Leu 15 10 15 egg tgg ggt ttt ctg tgt cat tet etc gag 77

Arg Trp Gly Phe Leu Cys His Ser Leu Glu 20 25- <210> 150 <211> 25 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400> 150

Ala Gin Gly Trp Cys Lys Glu Gly Pro Thr Leu Arg Glu Trp Leu Arg 15 10 15

Trp Gly Phe Leu Cys His Ser Leu Glu 20 25 <210> 151 <211> 837 <212> DNA <213>Artificial sequence <220><223> Synthetic peptide sequence <220>221> CDS <222> (57)..(785) <223> 70· 016775 1316961 <400> 151 tcgattaatc gatttgattc tagatttgtt ttaactaatt aaaggaggaa taacat atg 59

Met gac aaa act cac aca tgt cca cct tgt cca get ccg gaa etc ctg ggg 107 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 5 10 15 gga ccg tea gtc ttc etc ttc ccc cca aaa ccc aag gac Acc etc atg 155 Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 20 25 30 ate tee egg acc cct gag gtc aca tgc gtg gtg gig gac gtg age cac 203 lie Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 35 40 45 gaa gac cct gag gtc aag ttc aac tgg tac gtg gac ggc gtg gag gtg 251 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 50 55 60 65 cat aat gee aag Aca aag ccg egg gag gag cag tac aac age aeg tac 299 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr 70 75 80 cgt gtg gtc age gtc etc acc gtc ctg cac cag gac tgg ctg aat ggc 347 Arg Val Val Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn Gly 85 90 95 aag gag tac aag tgc aag gtc tee aac aaa gee etc cca gee ccc ate 395 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Lie 100 105 110 gag Aaa acc ate tee aaa gee aaa ggg cag ccc ega gaa cca cag gtg 443 Glu Lys Thr lie Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin Val 115 120 125 tac acc 'ctg ccc cca tee egg gat gag ctg acc aag aac cag Gtc age 491 Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gin Val Ser 130 135 140 145 ctg acc tgc ctg gtc aaa ggc ttc tat ccc age gac ate gee gtg gag 539 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp He Ala Val Glu 150 155 160 tgg gag age aat ggg cag ccg gag aac aac tac aag acc aeg cct ccc 587 Trp Glu Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 165 170 175 gtg ctg gac tee gac Ggc tee ttc ttc etc tac age aag etc acc gtg 635 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 180 185 190 gac aag age agg tgg cag cag ggg aac gtc ttc tea tgc tee gtg atg 683 016776 - 71 - 1316961

Asp Lys Ser Arg Trp Gin Gin Gly Asn Val Phe Ser Cys Ser Val Met 195 200 205 cat gag get ctg cac aac cac tac aeg cag aag age etc tee ctg tet 731 His Glu Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu Ser 210 215 220 225 ccg ggt aaa ggt gga ggt ggt ggt gca cag aaa geg gee gca aaa aaa 779 Pro Gly Lys Gly Gly Gly Gly Gly Ala Gin Lys Ala Ala Ala Lys Lys 230 235 240 etc gag taatggatcc gcggaaagaa gaagaagaag aagaaagccc gaaaggaagc tg 837 Leu Glu

<210> 152 <211> 243 <212> PRT <213> artificial sequence <220><223> synthetic peptide sequence <400>

Met Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 15 10 15

Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 20 25 30

Met lie Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 35 40 45

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 50 55 60

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr 65 70 75 80

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gin Asp Trp Leu Asn 85 90 95

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 100 105 110 -72- 1316961

He Glu Lys Thr lie Ser Lys Ala Lys Gly Gin Pro Arg Glu Pro Gin 115 120 125

Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gin Val 130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp lie Ala Val 145 150 155 160

Glu Trp Glu Ser Asn Gly Gin Pro Glu Asn Asn Tyr Lys Thr Thr Pro 165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 180 185 190

Val Asp Lys Ser Arg Τφ Gin Gin Gly Asn Val Phe Ser Cys Ser Val 195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gin Lys Ser Leu Ser Leu 210 215 220

Ser Pro Gly Lys Gly Gly Gly Gly Gly Ala Gin Lys Ala Ala Ala Lys 225 230 235 240

Lys Leu Glu <210> 153 <211> 45 <212>PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <220><221> miscjeature <222> 22)..(22) <223> At Position 22, PEG linker <400>(153

His lie Arg Glu Gly Pro Thr Leu Arg Gin Trp Leu Val Ala Leu Arg 15 10 15 -73 - 016778 1316961

Met Val Gly Gly Gly Pro Glu Gly Gly Gly Gly His lie Arg Glu Gly 20 25 30

Pro Thr Leu Arg Gin Trp Leu Val Ala Leu Arg Met Val 35 40 45 <210> 154 <211> 43 <212> PRT <213>Artificial Sequence <220><223> Synthetic Peptide Sequence <;220><221> misc_feature <222> (1)..(1) . <223> At Position 1, Fc at N-terminus <220><221>misc_feature<222> (43) ..(43) <223> At position 43, Fc on C-terminus <400> 154

Thr Cys Glu Gin Gly Pro Thr Leu Arg Gin Trp Leu Leu Cys Arg Gin 15 10 15

Gly Arg Gly Gly Gly Lys Gly Gly Gly Thr Cys Glu Gin Gly Pro Thr 20 25 30

Leu Arg Gin Trp Leu Leu Cys Arg Gin Gly Arg 35 40 <210> 155 <211> 40 <212> PRT <213>Artificial sequence<220>. <223> ritual peptide sequence <220 〉 <221> misc_feature -74- ·· 01f^77C) 1316961 <222> (1)..(1) <223> At position 1, Fc at N-terminus <400> 155

Gin Leu Gly His Gly Pro Thr Leu Arg Gin Trp Leu Ser Trp Tyr Arg 15 10 15

Gly Met Gly Pro Asn Gly Glu Leu Arg Ser Gly Pro Thr Leu Lys Glu 20 25 30

Trp Leu Val Trp Arg Leu Ala Gin 35 40 <210> 156 <211> 18 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <220><221>; misc-feature <222> (19)..(19) <223> At position 19, Fc at C-terminus <400>

Cys Ser Trp Gly Gly Pro Thr Leu Lys Gin Trp Leu Gin Cys Val Arg 15 10 15

Ala Lys <210> 157 <211> 25 <212> PRT < 213 > artificial sequence <220><223> synthetic winning sequence parameter. <220><221> miscjeature <222> (1), (1) <223> At position 1, Fc at N-terminus 016780 -75 - 1316961 <400> 157

Gly Gly Gly Lys Gly Gly Gly Ala Val Pro Gin Gly Pro Thr Leu Lys 15 10 15

Gin Trp Leu Leu Trp Arg Arg Cys Ala 20 25 <210> 158 <211> 18 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <220>;221> miscwfeature <222> (1)..(1) <223> At position 1, PEG group attached <400> 158

Cys Ser Ser Gly Gly Pro Thr Leu Arg Glu Trp Leu Gin Cys Arg Arg 15 10 15

Met Gin <210> 159 <211> 46 <212: · PRT < 213 > artificial sequence <220><223> synthetic peptide sequence <220><221> misc_feature <222> 1)..(1) <223> At position 1, Fc at N-terminus <400> 159

Gly Gly Gly Gly Gly Tyr Cys Asp Glu Gly Pro Thr Leu Lys Gin Trp 15 10 15 016781 -76- 1316961

Leu Val Cys Leu Gly Leu Gin Gly Gly Gly Gly Gly Tyr Cys Asp Glu 20 25 30

Gly Pro Thr Leu Lys Gin Trp Leu Val Cys Leu Gly Leu Gin 35 40 45 <210> 160 <211> 75 <212> PRT <213>Artificial Sequence<220><223> Synthetic peptide sequence <220> <221> misc_feature <222> (76)..(76) - <223> At position 76, Fc at C-terminus <400>

Cys Ser Trp Gly Gly Pro Thr Leu Lys Gin Trp Leu Gin Cys Val Arg 15 10 15

Ala Lys Gly Gly Gly Ala Gly Gly Gly Cys Ser Trp Gly Gly Pro Thr 20 25 30

Leu Lys Gin Trp Leu Gin Cys Val Arg Ala Lys Gly Gly Gly Ala Gly 35 40 45

Gly Gly Cys Ser Trp Gly Gly Pro Thr Leu Lys Gin Trp Leu Gin Cys 50 55 60

Val Arg Ala Lys Gly Gly Gly Ala Gly Gly Gly 65 70 75 <210> 161 <211> 43 4212> PRT <213>Artificial Sequence <220><223> Synthetic peptide sequence 016782 -77 - 1316961 <220><221> miscjeature <222> (44)..(44) <223> At position 44, PEG group at C-terminus <400>

Val Gly lie Glu Gly Pro Thr Leu Arg Gin Trp Leu Ala Gin Arg Leu 15 10 15

Asn Pro Gly Gly Gly Cys Gly Gly Gly Val Gly lie Glu Gly Pro Thr 20 25 30

Leu Arg Gin Trp Leu Ala Gin Arg Leu Asn Pro 35 40. <210> 162 <211> 40 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <220>;<221> misc_feature <222> (1)..(1) <223> At position 1, Fc at N-terminus <400>

Glu Leu Arg Ser Gly Pro Thr Leu Lys Glu Trp Leu Val Trp Arg Leu 15 10 15

Ala Gin Gly Gly Gly Gly Glu Leu Arg Ser Gly Pro Thr Leu Lys Glu 20 25 30

Trp Leu Val Trp Arg Leu Ala Gin 35 40 <210> 163 <211> 43 <212> PRT <213>Artificial sequence<220> 016783 1316961 <223> Synthetic peptide sequence <220>;221>misc-feature<222> (1)..〇) <223> At position 1, Fc at N-terminus <220><221> misc_feature <222>(44)..(44 ) <223> At position 44, Fc at C-terminus <400> 163

Ala Leu Arg Asp Gly Pro Thr Leu Lys Gin Trp Leu Glu Tyr Arg Arg 1 5 10 15

Gin Ala Gly Gly Gly Lys Gly Gly Gly Ala Leu Arg Asp Gly Pro Thr 20 25 30

u Arg Gin Ala 35 40 <210> 164 &lt 164

Ala Leu Arg Asp Gly Pro Thr Leu Lys Gin Trp Leu Glu Tyr Arg Arg 15 10 15

Gin Ala Ala Leu Arg Asp Gly Pro Thr Leu Lys Gin Trp Leu Glu Tyr 20 25 30

Arg Arg Gin Ala 35 <210> 165 <211> 36 <212> PRT <213> Artificial sequence 016784 79- 1316961 <220> <223> Synthetic peptide sequence <400>

Glu Ala Leu Leu Gly Pro Thr Leu Arg Glu Trp Leu Ala Trp Arg Arg 15 10 15

Ala Gin Glu Ala Leu Leu Gly Pro Thr Leu Arg Glu Trp Leu Ala Trp 20 25 30 .Arg Arg Ala Gin 35 <210> 166 <211> 36 <212> PRT <213>Artificial Sequence<220><223> Synthetic peptide sequence <400> 166

Ala Val Pro Gin Gly Pro Thr Leu Lys Gin Trp Leu Leu Trp Arg Arg 15 10 15

Cys Ala Ala Val Pro Gin Gly Pro Thr Leu Lys Gin Trp Leu Leu Trp 20 25 30

Arg Arg Cys Ala 35 <210> 167 <211> 36 <212>PRT <213> Artificial sequence <220>-<223> Synthetic peptide sequence <400>

Tyr Cys Asp Glu Gly Pro Thr Leu Lys Gin Trp Leu Val Cys Leu Gly 15 10 15 016785 -80- 1316961

Leu Gin Tyr Cys Asp Glu Gly Pro Thr Leu Lys Gin Trp Leu Val Cys 20 25 30

Leu Gly Leu Gin 35 <210> 168 <211> 36 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Cys Ser Ser Gly Gly Pro Thr Leu Arg Glu Trp Leu Gin Cys Arg Arg 15 10 15

Met Gin Cys Ser Ser Gly Gly Pro Thr Leu Arg Glu Trp Leu Gin Cys 20 25 30

Arg Arg Met Gin 35 <210> 169 <211> 36 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400> 169

Cys Ser Trp Gly Gly Pro Thr Leu Lys Gin Trp Leu Gin Cys Val Arg 15 10 15

Ala Lys Cys Ser Trp Gly Gly Pro Thr Leu Lys Gin Trp Leu Gin Cys 20 25 30

Val Arg Ala Lys 35 <210> 170 <211> 41 016786 •81 - 1316961 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400> 170

Ala Leu Arg Asp Gly Pro Thr Leu Lys Gin Trp Leu Glu Tyr Arg Arg 15 10 15

Gin Ala Gly Gly Gly Gly Gly Ala Leu Arg Asp Gly Pro Thr Leu Lys 20 25 30

Gin Trp Leu Glu Tyr Arg Arg Gin Ala 35 40 <210> 171 <211> 41 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Glu Ala Leu Leu Gly Pro Thr Leu Axg Glu Trp Leu Ala Trp Axg Arg 15 10 15

Ala Gin Gly Gly Gly Gly Gly Glu Ala Leu Leu Gly Pro Thr Leu Arg 20 25 30

Glu Trp Leu Ala Trp Arg Arg Ala Gin 35 40 <210> 172 <211> 41 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400> *172

Ala Val Pro Gin Gly Pro Thr Leu Lys Gin Trp Leu Leu Trp Arg Arg 15 10 15 -82- 016787 1316961

Cys Ala Gly Gly Gly Gly Gly Ala Val Pro Gin Gly Pro Thr Leu Lys 20 25 30

Gin Trp Leu Leu Trp Arg Arg Cys Ala 35 40 <210> 173 <211> 41 <212> PRT <21;3> Artificial sequence. <220><223> Synthetic peptide sequence <400>; 173

Tyr Cys Asp Glu Gly Pro Thr Leu Lys Gin Trp Leu Val Cys Leu Gly 15 10 15

Leu Gin Gly Gly Gly Gly Gly Tyr Cys Asp Glu Gly Pro Thr Leu Lys 20 25 30

Gin Trp Leu Val Cys Leu Gly Leu Gin 35 40 <210> 174 <211> 41 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <400>

Cys Ser Ser Gly Gly Pro Thr Leu Arg Glu Trp Leu Gin Cys Arg Arg 15 10 15

Met Gin Gly Gly Gly Gly Gly Cys Ser Ser Giy Gly Pro Thr Leu Arg 20 25 30

Glu Trp Leu Gin Cys Arg Arg Met Gin 35 40 -83 - 016788 1316961 <210> 175 <211> 41 <212>PRT <213>Artificial Sequence<220><223> Synthetic peptide sequence<;400> 175

Cys Ser Trp Gly Gly Pro Thr Leu Lys Gin Trp Leu Gin Cys Val Arg 15 10 15

Ala Lys Gly Gly Gly Gly Gly Cys Ser Trp Gly Gly Pro Thr Leu Lys 20 25 30

Gin Trp Leu Gin Cys Val Arg Ala Lys 35 40 <210> 176 <211> 23 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <220>221> misc_feature <222>(1)..(1) <223> At position 1, Fc at N-terminus <400> 176

Gly Gly Gly Gly Gly Ala Leu Arg Asp Gly Pro Thr Leu Lys Gin Trp 15 10 15

Leu Glu Tyr Arg Arg Gin Ala 20 <210> 177 <211> 23 <212> PRT : <213>Artificial sequence <220><223> Synthetic peptide sequence 016789 -84 - 1316961 <220&gt ; <221> misc_feature <222> (1)..(1) <223> At position 1, Fc at N-terminus <400> 177

Gly Gly Gly Gly Gly Glu Ala Leu Leu Gly Pro Thr Leu Arg Glu Trp 15 10 15

Leu Ala Trp Arg Arg Ala Gin 20 <210> 178 <211> 23 <212> PRT <213>Artificial sequence <220><223> Synthetic skin sequence <220>221> misc_feature <222> (1)..(1) <223> At position 1, Fc at N-terminus <400> 178

Gly Gly Gly Gly Gly Ala Val Pro Gin Gly Pro Thr Leu Lys Gin Trp 15 10 15

u Arg Cys Ala 20 <210><222> (1)..(1) <223> At position lt Fc at N-terminus -85 - 016790 1316961 <400> 179

Gly Gly Gly Gly Gly Tyr Cys Asp Glu Gly Pro Thr Leu Lys Gin Trp 1 5 10 15

Leu Val Cys Leu Gly Leu Gin 20 <210> 180 <211> 23 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <220><221> misc_feature <222> (1)..(1) -<223> At position 1, Fc at N-terminus <400> 180

Gly Gly Gly Gly Gly Cys Ser Ser Gly Gly Pro Thr Leu Arg Glu Trp 15 10 15

Leu Gin Cys Axg Arg Met Gin 20 <210> 181 <211> 23 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <220>221> misc_feature <222> (1)..(1) <223> At position 1, Fc at N-terminus <400> 181

Gly Gly Gly Gly Gly Cys Ser Trp Gly Gly Pro Thr Leu Lys Gin Trp 1 ' 5 10 15 016791 -86- 1316961

Leu Gin Cys Val Arg Ala Lys 20 <210> 182 <211> 46 <212> PRT < 2i3 > artificial sequence <220><223> synthetic peptide sequence <220><221> A feature <222> (1)..(1) <223> At position 1' Fc at N-terminus <400> 182

Gly Gly Gly Gly Gly Ala Leu Arg Asp Gly Pro Thr Leu Lys Gin Trp 15 10 15

Leu Glu Tyr Arg Arg Gin Ala Gly Gly Gly Gly Gly Ala Leu Arg Asp 20 25 30

Ly Arg Gin Ala 35 40 45 <210> 183 <211><220><221>misc-feature<222>(1)..(1)<223> At position 1, Fc at N-terminus <400> 183

Gly Gly Gly Gly Gly Glu Ala Leu Leu Gly Pro Thr Leu Arg Glu Trp 15 10 15

Leu Ala Trp Arg Arg Ala Gin Gly Gly Gly Giy Gly Glu Ala Leu Leu 20 25 30 016792 -87- 1316961

Gly Pro Thr Leu Arg Glu Trp Leu Ala Trp Arg Arg Ala Gin 35 40 45 <210> 184 <211> 46 <212> PRT <213> Artificial Sequence <220><2Z3> Synthetic peptide sequence <220><221> misc^feature <222> (1)., (1) <223> At position 1, Fc at N-terminus <400> 184 -

Gly Gly Gly Gly Gly Ala Val Pro Gin Gly Pro Thr Leu Lys Gin Trp 15 10 15

Leu Leu Trp Arg Arg Cys Ala Gly Gly Gly Gly Gly Ala Val Pro Gin 20 25 30

Gly Pro Thr Leu Lys Gin Trp Leu Leu Trp Arg Arg Cys Ala 35 40 45 <210> 185 <211> 46 <212> PRT <213>Artificial Sequence<220><223> Synthetic peptide sequence <220><221> miscjeature <222> (1)..(1) <223> At position 1, Fc at N-terminus <400>

Gly Gly Gly Gly Gly Tyr Cys Asp Glu Gly Pro Thr Leu Lys Gin Trp 15 10 15 -88- 016793 1316961

Leu Val Cys Leu Gly Leu Gin Gly Gly Gly Gly Gly Tyr Cys Asp Glu 20 25 30

Gly Pro Thr Leu Lys Gin Trp Leu Val Cys Leu Gly Leu Gin 35 40 45 <210> 186 <211> 46 <212> PRT <213>Artificial Sequence <220><223> Synthetic peptide sequence <220><221> misc-feature <222> (1)..(1) <223> At position 1, Fc at N-terminus <400>

Gly Gly Gly Gly Gly Cys Ser Ser Gly Gly Pro Thr Leu Arg Glu Trp 15 10 15

Leu Gin Cys Arg Arg Met Gin Gly Gly Gly Gly Gly Cys Ser Ser Gly 20 25 30

Gly Pro Thr Leu Arg Glu Trp Leu Gin Cys Arg Arg Met Gin 35 40 45 <210> 187 <211> 46 <212> PRT <213>Artificial Sequence<220><2Z3> Synthetic peptide sequence <220><221> misc^feature <222> (1)..(1) <223> At position 1, Fc at N-terminus <400> 187 *

Gly Gly Gly Gly Gly Cys Ser Trp Gly Gly Pro Thr Leu Lys Gin Trp 15 10 15 01679^ -89· 1316961

Leu Gin Cys Val Arg Ala Lys Gly Gly Gly Gly Gly Cys Ser Trp Gly 20 25 30

Gly Pro Thr Leu Lys Gin Trp Leu Gin Cys Val Arg Ala Lys 35 40 45 <210> 188 <211> 46 <212> PRT <213>Artificial Sequence<220><223> Synthetic peptide sequence <220><221> miscjeature <222> (47)..(47) - <223> At position 47> Fc at C-tenninus <400> 188

Ala Leu Arg Asp Gly Pro Thr Leu Lys Gin Trp Leu Glu Tyr Arg Arg 15 10 15

Gin Ala Gly Gly Gly Gly Gly Ala Leu Arg Asp Gly Pro Thr Leu Lys 20 25 30

Gin Trp Leu Glu Tyr Arg Arg Gin Ala Gly Gly Gly Gly Gly 35 40 45 <210> 189 <211> 46 <212> PRT <213>Artificial Sequence<220><223> Synthetic peptide sequence <220><221> miscjeature <222> (47)..(47) <223> At position 47, Fc at C-terminus <400> 189 0167Q*i -90· 1316961

Glu Ala Leu Leu Gly Pro Thr Leu Arg Glu Trp Leu Ala Trp Arg Arg 15 10 15

Ala Gin Gly Gly Gly Gly Gly Glu Ala Leu Leu Gly Pro Thr Leu Axg 20 25 30 .Glu Trp Leu Ala Trp Ai'g Arg Ala Gin Gly Gly Gly Gly 35 40 45 <210> 190 <211> 46 &lt ;212> PRT <21;3>Artificial sequence <220><223> Synthetic peptide sequence <220> - <221 <221 > misc-feature <222> (47)., (47) <223> At position 47, Fc at C-terminus <400> 190

Ala Val Pro Gin Gly Pro Thr Leu Lys Gin Trp Leu Leu Trp Arg Arg 15 10 15

Cys Ala Gly Gly Gly Gly Gly Ala Val Pro Gin Gly Pro Thr Leu Lys 20 25 30

Gin Trp Leu Leu Trp Arg Arg Cys Ala Gly Gly Gly Gly Gly 35 40 45 <210> 191 <211> 46 <212> PRT <213>Artificial Sequence<220><21j> Synthetic peptide sequence <220> ^ <221 > misc-feature <222> (47)..(47) <223> At position 47, Fc at C-terminus 016796 -91 - 1316961 <400>

Tyr Cys Asp Glu Gly Pro Thr Leu Lys Gin Trp Leu Val Cys Leu Gly 15 10 15

Leu Gin Gly Gy Gly Gly Gly Tyr Cys Asp Glu Gly Pro Thr Leu Lys 20 25 30

Gin Trp Leu Val Cys Leu Gly Leu Gin Gly Gly Gly Gly Gly 35 40 45 <210> 192 <211> 46 <212> PRT <213>Artificial Sequence<220><223> Synthetic peptide sequence · <220><221> misc_feature <222> (47)..(47) <223> At position 47, Fc at C-terrainus <400> 192

Cys Ser Ser Gly Gly Pro Thr Leu Arg Glu Trp Leu Gin Cys Arg Arg 15 10 15

Met Gin Gly Gly Gly Gly Gly Cys Ser Ser Gly Gly Pro Thr Leu Arg 20 25 30

Glu Trp Leu Gin Cys Arg Arg Met Gin Gly Gly Gly Gly Gly 35 40 45 <210> 193 <211> 46 <212> PRT <213>Artificial Sequence <220><223> Synthetic peptide sequence <220><221> misc_feature <222> (47)..(47) <223> At position 47, Fc at C-terminus • 92-016797 1316961 <400>

Cys Ser Trp Gly Gly Pro Thr Leu Lys Gin Trp Leu Gin Cys Val Arg 15 10 15

Ala Lys Gly Gly Gly Giy Gly Cys Ser Trp Gly Gly Pro Thr Leu Lys 20 25 30

Gin Trp Leu Gin Cys Val Arg Ala Lys Gly Gly Gly Gly Gly 35 40 45 <210> 194 <211> 23 <212> PRT <213>Artificial Sequence<220><223> Synthetic peptide sequence <220 <221 > misc-feature <222> (24)..(24) <223> At position 24, Fc at C-terminus <400>

Ala Leu Aig Asp Gly Pro Thr Leu L.ys Gin Trp Leu Glu Tyr Arg Arg 15 10 15

Gin Ala Gly Gly Gly GIv Gly 20 <210> 195 <211> 23 <212> PRT <213>Artificial sequence, i20><223> Synthetic peptide sequence <220><221> misc_feature <;222>(24)..(24)<223> At position 24, Fc at C-terminus 016798 •93 - 1316961 <400> 195

Glu Ala Leu Leu Gly Pro Thr Leu Ai'g Glu Trp Leu Ala Trp Arg Arg 1 5 10 15

Ala Gin Gly Gly Gly Gly Gly 20 <210> 196 <211> 23 <212> PRT <2i3> Artificial Sequence <220><223> Synthetic Peptide Sequence <220> <221> rnisc_feature <222> (24)..(24) · <223> At position 24, Fc at C-tenninus <400> 196

Glu Ala Leu Leu Gly Pro Thr Leu Arg Glu Trp Leu Ala Trp Arg 15 10 15

Ala Gin Gly Gly Gly Giy Gly 20 <210> 197 <211> 23 <212> PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <220><221> misc_featurc <222> (24)..(24) <223> Ai position 24, Fc at C-terminus <400> 197

Tyr Cys Asp Glu Gly Pro Thr Leu Lys Gin Trp Leu Val Cys Leu Gly I 5 10 15 016799 •94- 1316961

Leu Gin Gly Gly Gly Gly Gly 20 <210> 198 <2H> 23 <212> PRT <213>Artificial sequence<220><223> Synthetic peptide sequence <220><221> misc_feature <222> (24)..(24) <223> At position 24, Fc at C-teminus <400> 198

Cys Ser Ser Gly Gly Pro Thf Leu Arg Glu Trp Leu Gin Cys Arg Arg 15 10 15

Met Gin Gly Gly Gly Gly GLy 20 <210> 199 <211> 23 <212>PRT <213>Artificial sequence <220><223> Synthetic peptide sequence <220><221> misc_fsature <222> (24)..(24) <223> At position 24t Fc at C-teminus <400> 199

Cys Ser Trp Gly Gly Pro Thr Leu Lys Gin Trp Leu Gin Cys Val Arg 15 10 15

Aia Lys Gly GLy Gly Gly Gly 20 016800

Claims (1)

Π16璆®l123464|fe Specialist calf file - .43⁄4 Chinese application sample) Pick up, patent application range 1. A polynucleotide encoding a substance composition bound to an mpl receptor, wherein the composition is selected from A group consisting of the following formula: (Vl)v--(LNl)i--(TMPl)a--(LN2)m--(TMP2)b--(LN3)n--(TMP3)c --(LN4)0--(TMP4)d--(V2)w wherein TMP1, TMP2, TMP3 and TMP4 are each independently selected from the group consisting of sequence identification number: 1 to sequence identification number: 23, and a physiologically acceptable salt thereof; LN1' LN2, LN3 and LN4 are each independently a linker comprising a peptide; VI and V2 are each independently a vehicle comprising an Fc functional site; a, b, c; Each is independently an integer from 0 to 20, and at least one of a, bc, and d is not 0 • , 1, m, ! ! and 〇 are each an integer from 0 to 20; and '" and W are each an integer from 0 to 1, and at least one of v and w is not 0 〇 2. A performance vector, which is included in the application Polynuclear aluminate according to item 1 of the patent scope. 3. A host cell comprising a vector according to item 2 of the patent application. 4. The host cell according to the third aspect of the patent application, wherein the cell is an E. coli cell. 5. The host cell according to claim 3, wherein the cell is a prokaryotic cell. 81210-960726.doc 1316961 6. The host cell according to claim 3, wherein the cell is a eukaryotic cell. 81210-960726.doc -2-