TWI317281B - Specific binding agents of human angiopoietin-2 - Google Patents

Description

1317281 (1) 玖, invention description (the description of the invention should be stated: the technical field, prior art, content, implementation and schematic description of the invention) This application claims the 60th application of the US Provisional Application on October 11, 2001 The benefits of /328,624 are hereby incorporated by reference in their entirety. Scope of the Invention The present invention relates to a specific binding agent recognized and associated with angiopoietin-2 (Ang_2), and more particularly, the present invention relates to the production, diagnostic use and therapeutic use of the specific binding agent and its fragment. Its specific-ground and heart--2 knots. BACKGROUND OF THE INVENTION The formation of new blood vessels from existing blood vessels is necessary for the angiogenesis process. Usually, angiogenesis is regulated by 'angiogenic factors, but in the multi-physiological and pathological use of the disease is closely related to the pre- and anti-cases' like cancer, eye # from A ^ R clear neovascular disease , arthritis and psoriasis, this process may be carried out with each anvil*. Folkman, J., Nat. Med,, 1: 27-3 1 (1995). It is known that many diseases have an 'offence' with the control or unwanted angiogenesis effect. These diseases include, for example, neovascularization not limited to clear eyes, such as retinopathy (including diabetes, N: I disease, retinopathy), age-related macular degeneration, psoriasis, and vascular prosthesis. , hemangioma, atherosclerosis, inflammatory disease 'like rheumatoid or, ~% of inflammatory diseases, especially Guan Yin (including rheumatoid arthritis), FX; or other chronic inflammatory diseases, like It is chronic asthma, % pulse or post-transplantation π > implanted atherosclerosis, endometriosis and spinal disease, such as a solid tumor and fluid (or hematopoietic )swollen

SlVa-3⁄4 SSMm 1317281 (7) Tumors (like leukemia and lymphoma). Other diseases associated with unwanted angiogenesis will be apparent to those skilled in the art. Although regulation of angiogenesis has been implicated in many signal transduction systems, the most distinctive and dominant endothelial cell-selection system involves the Tie-2 receptor tyrosine kinase (called "Tie-2" or " Tie-2R" (also known as "ORK"); also refers to the mouse Tie-2 as "tek" and its ligand 'angiogenin (〇&16,虬\7· and Yancopoulos, GD ·, Genes Dev. 13: 1055- 1066 [1999]). There are 4 known angiopoietins; angiopoietin-1 (i'Ang-Γ) to angiopoietin-4 ("Ang-4"). These angiopoietins are also referred to as "Tie-2 ligands." (Davis, S. et al., Cell, 87: 1 161-1169 [1996]; Grosios, K. et al., Cytogenet

Cell Genet, 84: 118-120 [1999] ; Holash, J. et al., Investigative

Ophthalmology & Visual Science, 42: 1617-1625 [1999] ; Koblizek, TI et al, Current Biology, 8: 529-532 [1998]; Lin, P. et al., Proc Natl Acad Sci USA, 95: 8829- 8834 [1998] ; Maisonpierre, PC et al, Science, 277: 55-60 [1997] ; Papapetropoulos, A. et al, Lab Invest, 79: 213-223 [1999] ; Sato, TN et al, Nature, 375 : 70-74 [1998]; Shyu, KG et al., Circulation, 98: 2081-2087 [1998] ; Suri, C. et al., Cell, 87: 1171-1180 [1996] ; Suri, C. et al. Science, 282: 468-471 [1998] ; Valenzuela, DM et al., Proceedings of the National Academy of Sciences of the USA, 96: 1904-1909 [1999] * Witzenbichler, B. et al., J Biol Chem, 273: 18514-18521 [1998]). However, 'Ang-1 binds to Tie-2, and in the cultured endothelial cells' stimulation of receptor filling, Ang-2 agonism has been observed and antagonizes phosphorylation of Tie-2 receptor (Davis, S. et al. Person, [1996], former; Maisonpierre, PC et al., [1997], former;

1317281

Kim, I., JH Kim, et al, Oncogene 19(39): 4549-4552 (2000); Teichert-Kuliszewska, K., PC Maisonpierre et al, Cardiovascular Research 49(3): 659-70 (2001)) . The phenotypes of mouse Tie-2 and Ang-1 gene knockout are similar, and Ang-Ι stimulated Tie-2 phosphorylation is proposed to convey the developing blood vessels in the uterus by maintaining endothelial cell-supporting cell attachment. Transformation and stabilization (Dumont, DJ et al., Genes & Development, 8: 1897-1909 [1994]; Sato, T.N_ et al., Nature, 376: 70-74 [1995] ; Suri, C Et al., [1996], former). It is believed that the role of Ang-1 in vascular stabilization remains in adulthood, where it is widely and in composition (Hanahan, D. et al., Science, 277: 48-50 [1997]; Zagzag, D Wait for others,

Experimental Neurology, 159: 391-400 [1999]). Contrary to the performance of 'Ang-2', which is mainly limited to vascular remodeling, it is thought to block the function of _Ang-Ι, thereby inducing the state of vascular plasticity that contributes to angiogenesis (Hanahan, D. et al. , [1997], former; Holash, J., et al.

Science, 284: 1994-1998 [1999]; Maisonpierre, P.C. et al., [1997], former). Many published studies have probably demonstrated the performance of vascular-selective Ang-2 in disease states and are associated with angiogenesis. These pathological conditions include, for example, psoriasis, macular degeneration, and cancer (Bun〇ne, G. et al. 'American Journal of Pathology, 155: 1967-1976 [1999] Etoh, T. et al., Cancer Research, 61: 2145-2153 [2001] ; Hanga1' M. et al.

Investigative Ophthalmology & Visual Science' 42· 1617 162 [2001]; Holash. J. et al., [1999], supra; Kur〇da, K. et al., Journal 1317281 (4) of Investigative Dermatology, 1 16: 713-720 [2001] ; Otani, A. et al., Investigative Ophthalmology Sc Visual Science, 40: 1912-1920 [1999] : Stratmann,. A. et al., American Journal of Pathology, 153: 1459-1466 [1998] Tanaka, S. et al., J Clin Invest, 103: 34-345 [1999] ; Yoshida, Y. et al., International Journal of Oncology, 15: 1221-1225 [1999] ; Yuan, K. et al., Journal Of Periodontal Research, 35: 165-171 [2000]; Zagzag, D. et al., [1999], supra). Most of these studies focus on cancer, and many of these tumor types appear to exhibit vascular-Ang-2 performance. Contrary to its performance in pathological angiogenesis, Ang-2 is extremely restricted in normal tissues (Maisonpierre, PC et al., [1997], former; Mezquita, J. et al., Biochemical and Biophysical Research Communications, 260: 492-498 [1999]). In normal adults, the three major angiogenic locations are the ovary, placenta, and uterus; these are the primary groups in normal (ie, non-cancerous) tissues in which Ang-2 mRNA has been detected. Weaving. Some functional studies suggest that Ang-2 may be involved in tumor angiogenesis. Ahmad et al. (Cancer Res., 61: 1255-1259 [2001]) describe the overexpression of Ang-2 and suggest that it may be associated with an increase in tumor growth in a mouse xenograft mode. See also Etoh et al., formerly and Tanaka et al., previously submitted information that may correlate Ang-2 overexpression with tumor vascular hyperplasia. However, in contrast, Yu et al. (Am. J. Path) .158: 563-570 [2001]) The reported data show that Ang-2 is overexpressed in Lewis lung cancer and TA3 breast cancer cells, possibly prolonging the survival of mice injected with the corresponding transfectant. 1317281 (5) Over the past few years, various publications have suggested Ang-1, Ang-2 and/or Tie-2 as potential targets for anticancer therapy. For example, U.S. Patent Nos. 6,166,185, 5,650,490 and 5,814,464, respectively, disclose the concept of anti-Tie-2 ligand antibodies and receptor bodies. Lin et al. (Proc. Natl. Acad. Sci USA, 95: 8829-8834 [1998]) injected soluble Tie-2 expressed by adenovirus into mice; soluble Tie-2 may reduce the number of tumors developed in mice and size. In a related study, Lin et al. (J. Clin. Invest., 100: 2072-2078 [1997]) injected a soluble form of Tie-2 into rats; the compound may be lowered. The size of the tumor. Siemeister et al. (Cancer Res., 59: 3 185-3 189 [1999]) produced human melanoma cell lines expressing the extracellular functional sites of Tie-2, which were injected into nude mice and identified as soluble. Tie-2 may cause "significant inhibition" of tumor growth and tumor angiogenesis. Reviewing this information and getting both Ang-1 and Ang-2 to bind to Tie-2, from these studies, it is not clear whether Ang-1, Ang-2 or Tie-2 will be attractive for anti-cancer therapy. The target of force. The PCT publication WO 00/24782, published on May 4, 2000, describes the fusion of certain peptides with stable plasma proteins, such as Ig constant regions, to improve the half-life of these molecules. The fusion of proteins or fragments thereof with stable plasma proteins, such as Ig constant regions, has been described in many ways to improve the half-life of these molecules (see, e.g., U.S. Patent No. 5,480,981; Zheng et al., J. Immunol., 154: 5590-5600, (1995); Fisher et al, Ν Engl. J. Med., 334: 1697-1702, (1996); Van Zee, K. et al., J. Immunol., 156: 2221- 2230, (1996); U.S. Patent No. 5,808,029 issued on September 15, 1998; Capon et al.-10-1317281 (6) Fen Meng, Nature, 337: 525-531, (1989): Harvill et al. I_unotech, 1: 95-105, (1995); published on July 3, 1997, w〇97/23614; applied on December 11, 1997? <: Ding/1^97/23183;;1^1 Qiao, 1. Take 1).1^(1,, 174: 561-569, (1991); w〇95/21258, published on August 10, 1995).

Effective anti-Ang-2 treatment may benefit large populations of cancer patients, as most solid tumors require new blood vessel formation to grow to a diameter of 1-2 mm or more. Such treatments may have a wide range of applications in other diseases associated with angiogenesis, such as retinopathy, arthritis, and porcine sputum. There is an undeveloped need for new formulations that are specifically recognized and integrated with Ang-2. Such formulations will be useful for diagnostic screening and for the treatment of disease states associated with Ang-2 activity. Accordingly, it is an object of the present invention to provide a specific binding agent for Ang-2 which modulates Ang-2 activity. Such preparations of the invention are in the form of peptibodies, i.e., peptides fused to other molecules, such as the Fc functional site of an antibody, wherein the peptide moiety specifically binds to Ang-2.

SUMMARY OF THE INVENTION The invention is indicated in the specific examples of peptides (also referred to herein as polypeptides) that bind to Ang-2. Variants and derivatives of such peptides are also included in the present invention. In other specific embodiments, the peptides of the invention and variants and derivatives thereof are mediated to the mediator. In other embodiments, the peptide can be fused to an Fc functional site, thereby providing a peptibody. Including at least one, for example, sequence recognition: -11 - 1317281 __. No. 3 to sequence identification: No. 6, or sequence identification: 76 to sequence identification: peptide number ι57, as well as variants and derivatives thereof. In addition, the peptide may also include at least one peptide according to the formula set forth in Sequence Recognition: No. 65 to Sequence Recognition: No. 75 and Sequence Identification: No. 158. In another embodiment, the invention provides nucleic acid molecules encoding the specific binding agents, as well as variants and derivatives thereof. In another embodiment, the invention provides nucleic acid molecules' encoding propeptides, as well as variants and derivatives thereof. Such nucleic acid molecules may include sequence identification as needed: number 33 to sequence identification: number 53. In another embodiment, the invention provides a method of reducing tumors by administering to a subject in need thereof an effective amount of a specific binding agent of the invention. The invention also provides a method of inhibiting angiogenesis in an individual comprising administering to the individual in need thereof an effective amount of a specific binding agent of the invention. The invention further provides a method of treating cancer in an individual comprising administering to the individual in need thereof an effective amount of a specific binding agent of the invention. The invention also relates to a polymorphism capable of binding to Ang-2, the tangyl T 13⁄4 polypeptide comprising The amino acid sequence WDPWT (sequence recognition: No. 65), the length of the polypeptide of the Japanese kansui 々 is from 5 to 50 amino acids' and its physiologically acceptable salts. The polypeptide may also include an amino acid sequence:

WDPWTC (sequence recognition: No. 66) and its physiologically acceptable salts. Furthermore, the polypeptide may comprise an amino acid sequence:

Cz2WDPWT (SEQ ID NO: 67) -12· 1317281 (8) wherein Z2 is an acidic or neutral polar amino acid residue, and a physiologically acceptable salt thereof. The polypeptide may also include an amino acid sequence:

Cz2WDPWTC (SEQ ID NO: 68) wherein z2 is an acidic or neutral polar amino acid residue, and a physiologically acceptable salt thereof. In other specific embodiments, the invention relates to a polypeptide capable of binding to Ang-2, comprising an amino acid sequence of the formula: akYCaSWDPWTCaW14 (SEQ ID NO: 69) wherein: a1, a2 and a3 are each an amine group Acid residue; a5 is an amino acid residue; _ a12 is absent or an amino acid residue; a13 lacks a neutral hydrophobic, neutral polar, or basic amino acid residue; a14 is A neutral hydrophobic, or neutral, polar amino acid residue: and a physiologically acceptable salt thereof. In a preferred embodiment: a1 is V, I, P, W, G, S, Q, N, E, K, R or H; a2 is V, P, M, G, S, Q, D , E' K, R or Η; a3 is Α, V, P, M, F, T, G, D, Ε, Κ or Η; a5 is A, V, G, Q, Ν 'D or Ε; a12 Is S, Q, N, D, E, K or R; a13 is L, T or H; and-13-

Production I ·, _ ·ι·^··ι. —If _ mmMM 1317281 (9) a14 is V, L, I, W or M. In a more preferred embodiment, a1 is Q; a2 is E; a3 is E: a5 is D or E; a12 is D or E; a13 is H; and a14 is M. It should be understood that the use of lowercase letters with shoulder numbers (such as a1 and bb, in this paper, attempts to confirm the position of the amino acid, rather than a single letter abbreviation intended to represent a particular amino acid. In this article, a single letter is provided in capital letters. Amino acid abbreviation β

The invention further relates to a polypeptide capable of binding to Ang-2, comprising an amino acid sequence of the formula: bVbVbVCbSWDPWTCb'VVVVV0 (SEQ ID NO: 70) wherein: bl is absent or an amino acid residue; b2 is absent or medium a hydrophobic, neutral polar or basic amino acid residue; b3, b4, b5 and b6 are respectively absent or an amino acid residue; b8 is an amino acid residue;

B15 lacks or amino acid residues; b16 lacks or neutral hydrophobic, neutral polar or basic amino acid residues; b17 lacks either neutral or neutral polar amine groups Acid residues; b18, b19 and b2G are respectively absent or amino acid residues;

And its physiologically acceptable salts. In a preferred embodiment: b1 is missing or A, V, L, P, W, F, T, G, S' Q, N, K -14-1317281 (ίο) b2 is missing or A, V , L, I, P, W, Μ, T, G, S, Y, N, Κ, R or Η; b3 is missing. or .A, L ' I ' P, W, M, T, G, S , Q, N, ER or Η; b4 is V, I ' P, W, G, s, Q, N, E, K, R or H; b5 is V, P, M, G, S' Q, D , E, K ' R or H ; b6 is A, V, P, M, F, T, G, D, 'E, K4 ' H; b8 is A, V, G, Q, N, D or E; B15 is r S' , Q , N , D , E , or R r b16 is L, Τ or Η; b17 is V, L, I, W or Μ;

B18 is missing or A, V, L, P, W' F, T, G, Y, Q, D, E or R; Top 19 is missing or ¥, 1^, 1,? , D, 0, 5, 丫, (5, :^, 0, [ or R; and b20 is missing or V, L, P, W, Μ, T, G, S, Y, Q, N, D, Ruler or R. In a more specific embodiment, b1 is absent or P or T; b2 is absent or I or N: b3 is absent or R or I; b4 is Q; b5 is E; b6 is E; b8 Is D or E; b15 is D or E; b16 is Η; b17 is Μ; b18 is absent or is W or P; b19 is absent or is G or E; and b2Q is absent or is V or K. Preferably, the polypeptide comprises at least one amino acid sequence selected from the group consisting of sequence recognition: No. 4 and sequence recognition: No. 76 to sequence identification: No. 118, wherein the polypeptide is capable of binding to Ang-2, and Physiologically acceptable salts. The peptide sequence is set forth below: -15- 1317281 (ίο

Table 1 Peptide sequence identification number Peptide sequence Con4-44 76 PIRQEECDWDPWTCEHMWEV Con4-40 77 TNIQEECEWDPWTCDHMPGK Con4-4 78 WYEQDACEWDPWTCEHMAEV Con4-31 79 NRLQEVCEWDPWTCEHMENY Con4-C5 80 AATQEECEWDPWTCEHMPRS Con4-42 81 LRHQEGCEWDPWTCEHMFDW Con4-35 82 VPRQKDCEWDPWTCEHMYVG Con4-43 83 sisheecewdpwtcehmqvg Con4- 49 84 WAAQEECEWDPWTCEHMGRM Con4-27 .. 85 TWPQDKCEWDPWTCEHMGST Con4-48 86 GHSQEECGWDPWTCEHMGTS Con4-46 87 QHWQEECEWDPWTCDHMPSK Con4-41 88 NVRQEKCEWDPWTCEHMPVR Con4-36 89 KSGQVECNWDPWTCEHMPRN Con4-34 90 VKTQEHCDWDPWTCEHMREW Con4-28 91 AWGQEGCDWDPWTCEHMLPM Con4-39 92 PVNQEDCEWDPWTCEHMPPM Con4-25 93 RAPQEDCEWDPWTCAHMDIK Con4-50 94 HGQNMECEWDPWTCEHMFRY Con4-38 95 PRLQEECVWDPWTCEHMPLR Con4-29 96 RTTQEKCEWDPWTCEHMESQ Con4-47 97 QTSQEDCVWDPWTCDHMVSS Con4-20 98 QVIGRPCEWDPWTCEHLEGL Con4-45 99 WAQQEECAWDPWTCDHMVGL Con4-37 100 LPGQEDCEWDPWTCEHMVRS Con4-33 101 PMNQVECDWDPWTCEHMPRS AC2-Con4 102 FGWSHGCEWDPWTCEHMGST Con4-32 103 KSTQDDCDWDPWTCEHMVGP Con4-17 104 GPRISTCQWDPWTCEHMDQL Con4-8 105 STIGDMCEWDPWTCAHMQVD AC4-Con4 106 VLGGQGCEWDPWTCRLLQGW Con4,l 107 VLGGQGCQWDPWTCSHLEDG

-16- Invention of 萌萌 Ί • BaeevseHBBtB^BK» 1317281 (12)

Con4-Cl 108 TTIGSMCEWDPWTCAHMQGG Con4-21 109 TKGKSVCQWDPWTCSHMQSG Con4-C2 110 TTIGSMCQWDPWTCAHMQGG Con4-18 ..111 WVNEVVCEWDPWTCNHWDTP Con4-19 112 WQVGMCQWDPWTCKHMRLQ Con4-16 113 AVGSQTCEWDPWTCAHLVEV Con4-ll 114 QGMKMFCEWDPWTCAHIVYR Con4-C4 115 TTIGSMCQWDPWTCEHMQGG Con4-23 116 TSQRVGCEWDPWTCQHLTYT Con4-15 117 QWSWPPCEWDPWTCQTVWPS Con4-9 118 GTSPSFCQWDPWTCSHMVQG TN8-Con4* 4 QEECEWDPWTCEHM It should be understood that certain peptides and/or peptides may contain the prefix "TN", "TN8" or "TN12" and may or may not provide a specific peptibodies. The prefix. Thus, for example, "TN8-Con4" and "Con4"-words may be used interchangeably herein. In another embodiment, the invention relates to a composition having a substance of the formula: (X1)a-F1 -(X2)b and its multimer, wherein: F1 is the medium: 'X1 and X2 are respectively selected from -(L^c-P1; -(L1)c-Pl-(L2)d-P2; -(L1 c-P1-(L2)d-P2-(L3)e-P3 ; ^ -(L^e-P'^L^d-P'-CL^e-P'-iL^rP4 ; one or The plurality of P1, P2, P3 and P4 respectively comprise a polypeptide as described herein. For example, in a preferred embodiment, P1, P2, P3 and 13172281 (13) P4 may comprise sequence recognition, respectively: number 3 to Sequence recognition: No. 6, and/or sequence recognition: No. 76 to sequence identification: polypeptide of No. 157. In another specific embodiment, the composition of the substance is of the formula: X'-F1 or F[-X2 and Physiologically acceptable salts, wherein X1, F1 and X2 are as defined herein. In another embodiment, the composition of matter is of the formula: -F^CL1), -?1 and a permissible salt, wherein L1, F1 and P1 are as defined herein. In another embodiment In the example, the compound of the substance is a formula F1-(L1)c-P1-(L2)d-P2 and a physiologically acceptable salt thereof, wherein L1, F1, P1, P2 and c and d, As defined herein, in another embodiment, the composition of matter is of the formula: P'-CL^^F^CL^a-P2 and its physiologically acceptable salts. In a specific embodiment, F1 is an Fc functional site or a fragment thereof. The invention more relates to a polypeptide capable of binding to Ang-2, comprising an amino acid sequence of the formula:

Pc2Dc4Lc6c7c8LY (SEQ ID NO: 71) wherein c2 is a neutral hydrophobic amino acid residue; -18-». |· I · ll _ I _ hair 1317281 (14) c4 is A, D or E; c6 is An acidic amino acid residue; C7 is an amino acid residue; and C8 is a neutral hydrophobic, neutral polar or basic amino acid residue; and a physiologically acceptable salt thereof . In a preferred embodiment, c2 is L or Μ. In another preferred embodiment, c6 is D or E. The invention further relates to a polypeptide capable of binding to Ang-2, comprising an amino acid sequence of the formula:

ά'(12(13ά4Ρά6Οά8Ε(110(111(1ι^Υά15ά16(117(ί18(ί19ά20(121(22))), where d1 is absent or an amino acid residue; d2 is absent or neutrally polar, An acidic or basic amino acid residue; d3 lacks a neutral or neutral polar amino acid residue; d4 is absent or an amino acid residue; d6 is a neutral hydrophobic amine Base acid residue; d8 is A, D or E; · d1Q is an acidic amino acid residue; d" is an amino acid residue; d12 is a neutral hydrophobic, neutral polar or basic amine Base acid residue; d15 lacks a neutral or acidic, acidic or ageing amino acid residue; d16 lacks a neutral or acidic, acidic or basic amino acid residue; d17 is absent or Is a neutral hydrophobic or neutral polar amino acid residue; d18 lacks a neutral or neutral polar amino acid residue; • 19-1317281 (15) d19 lacks or is neutral Hydrophobic, neutral polar or basic amino acid residue; £12 () lacks or amino acid residue; d21 lacks or neutral polar, acidic or basic amine Acid residue; d22 lacks a neutral or hydrophobic, neutral polar or basic amino acid residue; and a physiologically acceptable salt thereof. In a preferred embodiment: d1 Is T, S, Q, R or Η; d2 is Τ, Q, Ν or Κ; d3 is F; d4 is Μ, Q, E or K; d6 is L or Μ; d8 is D or Ε; d10 is E D11 is Q or E; d12 is T or R; d15 is Y, D, E or K; d16 is Q: d17 is W or F: d18 is L, I, Μ or T; d19 is L, F or Υ D2() is Q, D or E; d21 is absent or Q or Η; d22 is absent or A, L, G, S or R. · -γ- -=-^ -i. -τ·;" n i-TTji^a-fcrya .a 1317281 (16) In a preferred embodiment, the polypeptide comprises at least one selected from the group consisting of sequence recognition: number 6 and sequence recognition: 119 to sequence identification: 142 a group of amino acid sequences wherein the polypeptide is capable of binding to Ang-2. Sequence identification is set forth below: number 6 and sequence recognition: 1 19-142:

Peptide SEQ ID NO Peptide Sequence L1-1 119 QNYKPLDELDATLYEHFIFHYT L1-2 120 LNFTPLDELEQTLYEQWTLQQS L1-3 121 TKFNPLDELEQTLYEQWTLQHQ L1-4 122 VKFKPLDALEQTLYEHWMFQQA L1-5 123 VKYKPLDELDEILYEQQTFQER L1-7 124 TNFMPMDDLEQRLYEQFILQQG L1-9 125 SKFKPLDELEQTLYEQWTLQHA L1-10 126 QKFQPLDELEQTLYEQFMLQQA L1-11 127 QNFKPMDELEDTLYKQFLFQHS L1-12 128 YKFTPLDDLEQTLYEQWTLQHV L1-13 129 QEYEPLDELDETLYNQWMFHQR L1-14 130 SNFMPLDELEQTLYEQFMLQHQ L1-15 131 QKYQPLDELDKTLYDQFMLQQG L1-16 132 QKFQPLDELEETLYKQWTLQQR L1-17 133 VKYKPLDELDEWLYHQFTLHHQ L1-18 134 QKFMPLDELDEILYEQFMFQQS L1-19 135 QTFQPLDDLEEYLYEQWIRRYH L1-20 136 EDYMPLDALDAQLYEQFILLHG L1-21 137 HTFQPLDELEETLYYQWLYDQL L1-22 138 YKFNPMDELEQTLYEEFLFQHA AC6-L1 139 TNYKPLDELDATLYEHWILQHS L1-C1 140 QKFKPLDELEQTLYEQWTLQQR L1-C2 141 TKFQPLDELDQTLYEQWTLQQR L1-C3 142 TNFQPLDELDQIXYEQWTLQQR L1 6 KFNPLDELEETLYEQFTFQQ

-21 - 1317281 (9) The present invention also relates to a polypeptide capable of binding to Ang-2, comprising an amino acid sequence of the formula:

RPe3e4e3e6e7G (SEQ ID NO: 73) wherein e3 is a neutral polar amino acid residue; e4 is an acidic amino acid residue; e5 is a neutral polar or acidic amino acid residue;

E6 is a neutral hydrophobic amino acid residue; e7 is a neutral hydrophobic amino acid residue; and a physiologically acceptable salt thereof. In a preferred embodiment, e3 is Y or Ο. In another preferred embodiment, e4 is D or E. In another preferred embodiment, e6 is I or Μ. The invention further relates to a polypeptide capable of binding to Ang-2, comprising an amino acid sequence of the formula: flf2f3£4&pf7f8f9fl0fllQfl3fl4fl5fl6fl7fl8fl9f20 (SEQ ID NO: 74) wherein f1 is a neutral hydrophobic or neutral polar amine Acidic acid residue; f2 is a neutral hydrophobic or neutral polar amino acid residue; f3 is a neutral polar or acidic amino acid residue; f4 is neutral hydrophobic or neutral polar Amino acid residue; f7 is a neutral polar amino acid residue; f8 is an acidic amino acid residue; -22-

Mt circle - I g ·»^ρΐ Inventive hybrid 1317281 〇s) f9 is a neutral polar or acidic amino acid residue; [1 (5 is a neutral hydrophobic amino acid residue; f11 is Neutral hydrophobic amino acid residue; f13 is a neutral hydrophobic or neutral polar amino acid residue; f14 is a neutral hydrophobic or neutral polar amino acid residue; f15 is medium a polar amino acid residue; f16 is a neutral polar amino acid residue; f17 is a neutral polar or acidic amino acid residue; f18 is a neutral hydrophobic or basic amino acid Residue; f19 is a neutral hydrophobic or neutral polar amino acid residue; and f2Q is a neutral hydrophobic or neutral polar amino acid residue; and a physiologically acceptable salt thereof In a preferred embodiment: f1 is S, A or G; f2 is G, Q or P; f3 is Q ' G or D; f4 is L, Μ or Q; f7 is C or Υ; f8 E or D; f9 is E, G or D; f1() is I or Μ; f11 is F or L; f13 is C or W; f14 is G or P; f15 is T or N; -23- 1317281

MUSS (19) f16 is Q, Y or K; f17 is N, D or Q; f18 is L, V, W or R; f19 is A, Q, Y or I: and f2G is L, A, G or V .

In a more preferred embodiment, the invention relates to a population comprising at least one polypeptide selected from the group consisting of sequence recognition: number 3 and sequence recognition: 143 to sequence identification: 148 amino acid sequence, wherein the polypeptide It is capable of binding to Ang-2 and its physiologically acceptable salts. Sequence identification: No. 3 and sequence identification: No. 143 to sequence identification: No. 148 is as follows.

Peptide SEQ ID NO: 143 AGGMRPYDGMLGWPNYDVQA Con 1-2 144 QTWDDPCMHILGPVTWRRCI Coni-3 145 APGQRPYDGMLGWPTYQRIV Con 1-4 146 SGQLRPCEEIFGCGTQNLAL Coni-5 147 FGDKRPLECMFGGPIQLCPR Con 1-6 148 GQDLRPCEDMFGCGTKDWYG Coni 3 KRPCEEBFGGCTYQ

In other aspects, the invention relates to a polypeptide that binds to Ang-2, comprising an amino acid sequence of the formula:

Cg2Gg4g5DPFTg10GCg13 (Sequence recognition: No. 75) wherein g2 is an acidic amino acid residue; -24- 1317281 wicking said resuscitation phase (20) g4 is a neutral hydrophobic amino acid residue; g5 is E, D Or Q: Giant 10 is a neutral hydrophobic or neutral polar amino acid residue; g13 is an acidic residue: and a physiologically acceptable salt thereof. In a preferred embodiment, g2 is E or D. In another preferred embodiment, g4 is V or Μ. In another embodiment, Lu 1 (5 is F or Q. In another embodiment, g13 is D or deuterium.

The invention further relates to a polypeptide capable of binding to Ang-2, comprising an amino acid sequence of the formula: h1h2h3h4Ch6Gh8h9DPFTh14GCh17hI8h19h20 (SEQ ID NO: 158) wherein h1 is absent or neutrally hydrophobic, neutrally polar or alkaline Amino acid residues: h2 is a neutral hydrophobic or neutral polar amino acid residue: h3 is an acidic amino acid residue; · h4 is a neutral hydrophobic or neutral polar amino acid Residue; h6 is an acidic amino acid residue: h8 is a neutral hydrophobic amino acid residue; h9 is E, D or Q; h14 is a neutral hydrophobic or neutral polar amino acid residue H17 is an acidic amino acid residue; h18 is a neutral hydrophobic, neutral polar or a test amino acid residue; h19 is a neutral hydrophobic or neutral polar amino acid residue Base; and •25- 1317281

(21) h2() is absent or an amino acid residue; and a physiologically acceptable salt thereof. In a preferred embodiment: h1 is absent or A, L, M, G, K or H; h2 is L, F or Q; h3 is D or E; h4 is W or Y; h6 is D or E H8 is V or Μ; h14 is F or Q; h17 is D or E; h18 is Μ, Y, N or K: h19 is L or Q: and h20 is absent or Μ, T, G, S, D, K or R. In a more preferred embodiment, the invention relates to a population comprising at least one polypeptide selected from the group consisting of: sequence recognition: number 5 and sequence recognition: number 149 to sequence identification: 157 amino acid sequence, wherein the polypeptide The ability to bind to Ang-2, and its physiologically acceptable salts, is set forth below for sequence recognition: number 5 and sequence recognition: number 149 to sequence identification: number 157.

Peptide SEQ ID NO: 12-9-1 149 GFEYCDGMEDPFTFGCDKQT 12-9-2 150 KLEYCDGMEDPFTQGCDNQS 12-9-3 151 LQEWCEGVEDPFTFGCEKQR 12-9-4 152 AQDYCEGMEDPFTFGCEMQK 12-9-5 153 LLDYCEGVQDPFTFGCENLD 12-9-6 154 HQEYCEGMEDPFTFGCEYQG 12- 9-7 155 MLDYCEGMDDPFTFGCDKQM 12-9-C2 156 LQDYCEGVEDPFTFGCENQR 12-9-C1 157 LQDYCEGVEDPFTFGCEKQR 12-9 5 FDYCEGVEDPFTFGCDNH -26- 1317281 — ' (22) In a very preferred embodiment, the invention relates to having the following formula a composition of matter: (Xl)q-¥l-(X2)r and its multimer, wherein: F1 is a medium; X1 and X2 are respectively selected from m; -(Lbs-PMl^VPyLiu-P3; and -( L1)s-P1-(L2)t-P2-(L3)u-P3-(L4)v-P4; wherein one or more of P1, P2, P3 and P4 respectively comprise a polypeptide selected from the group consisting of : (a) Amino acid sequence WDPWT (sequence recognition: No. 65), wherein the length of the polypeptide is from 5 to 50 amino acids; (b) amino acid sequence WDPWTC (sequence recognition: No. 66); Amino acid sequence Cz2WDPWT (sequence recognition: No. 67), wherein z2 is an acidic or neutral polar amino acid residue; (d) amino acid sequence Cz2WDPWTC (SEQ ID NO: 68), wherein z2 is an acidic or neutral polar amino acid residue: (e) amino acid sequence Pc2Dc4Lc6c7c8LY (SEQ ID NO: 71), wherein c2 is a neutral hydrophobic amine group Acid residue; c4 is A, D or E; c6 is an acidic amino acid residue; c7 is an amino acid residue; and c8 is a neutral hydrophobic, neutral polar or basic amino acid (f) amino acid sequence RPe3e4e5e6e7G (SEQ ID NO: 73), wherein e3 -27-1317281 (23) is a neutral polar amino acid residue; e4 is an acidic amino acid residue; e5 a neutral or acidic amino acid residue: e6 is a neutral hydrophobic amino acid residue; and e7 is a neutral hydrophobic amino acid residue; (g) amino acid sequence Cg2Gg4g5DPFTg10GCg13 (Sequence recognition: No. 75), wherein g2 is an acidic amino acid residue; g4 is a neutral hydrophobic amino acid residue: g5 is a neutral polar or acidic amino acid residue: g1() a neutral hydrophobic or neutral polar amino acid residue; and g13 is an acidic residue; (h) sequence recognition: polypeptide number 1; (i) sequence recognition: number 2 And (j) sequence recognition: polypeptide of number 7; wherein L1, L2, L3, and L4 are junctions, respectively; and q, r, s, t, u, and v are 0 or 1, respectively, with a constraint of q and At least one of r is 1; and its physiologically acceptable salt. It will be appreciated that the invention further relates to fusion polypeptides comprising at least one peptide and vehicle as described herein, wherein the fusion polypeptide is capable of binding to Ang-2, and a physiologically acceptable salt thereof. In the fusion polypeptide, the vector is preferably at least one Fc functional site, polyethylene glycol, lipid, cholesterol group, carbohydrate and oligosaccharide. Those skilled in the art will appreciate that other suitable vehicles, such as albumin and the like, are also included within the scope of the invention. Those skilled in the art will recognize that various molecules can be inserted into the structure of a specific binder. Thus, a particular molecule can be inserted between, for example, the peptide and the media portion of the specific binding agent, or inserted within the peptide portion itself while still retaining the desired activity of the specific binding agent. It is easy to insert, for example, a molecule such as an Fc functional site or a fragment thereof, a polyethylene glycol or other related molecule, such as a Portuguese -28-1317281 hair (24)

Glycans, fatty acids, lipids, cholesterol groups, small carbohydrates, peptide cytotoxic preparations, chemotherapeutic agents, detectable moieties as described herein (including fluorescent agents, radioactive labels, such as radioisotopes), and Oligosaccharides, oligonucleotides, polynucleotides, interference (or other) RNA, enzymes, hormones or the like. Those skilled in the art will recognize other molecules suitable for insertion in this manner and are included within the scope of the invention. This includes, for example, the insertion of the desired molecule between two consecutive amino acids, optionally via a suitable linker. For example, in the Con4(C) peptidic sequence: M-Fc-GGGGGAQQEECEWDPWTCEHMLE (sequence recognition: No. 23) Those skilled in the art can easily remnant, for example, two adjacent glutamine ("QQ") The desired molecule is inserted between the bases to obtain the desired structure and/or function while still retaining the ability of the peptide to bind to Ang-2. Therefore, the sequence can be modified as follows: M-Fc-GGGGGAQ-[Molecule]-QEECEWDPWTCEHMLE An appropriate linker molecule can be added if necessary. It is further known that the molecule can be inserted at a number of positions on the molecule, including on the appropriate side chain, between the vector and the peptide sequence, as follows: Spring M-Fc-[Molecule]-GGGGGAQQEECEWDPWTCEHMLE or at any skilled person Other locations you want. Those skilled in the art should be aware of other suitable embodiments. In another embodiment, the invention is directed to a polynucleic acid that encodes a specific binding agent (including but not limited to peptides and/or peptibodies) of the invention as described herein. Those skilled in the art should understand that where known amino acid sequences are known, the corresponding nucleotide sequences can be easily determined using known techniques. -29- » -p»' — »π«» 1317281 (25). See, for example, Suzuki, D., An Introduction to Genetic Analysis, W. H. Freeman Pub. Co. (1986). Representative nuclear sinus acid sequences encoding the skin of the present invention are set forth below. Those skilled in the art will recognize that more than one codon can encode a particular amino acid' and thus the present invention relates to any peptide encoding the present invention. And/or the nuclear station acid sequence of the peptibodies. Representative peptide sequence SEQ ID NO DNA sequence Con4-44 76 PIRQEECDWDPWTCEHMWEV cc gatcc gtcaggaagaatgc ga ctgggacccgtggacctgcgaac acatgtgggaagtt (^ identification sequence: No. 159) Con4-40 77 TNIQEECEWDPWTCDHMPGK accaacatccaggaagaatgcga atgggacccgtggacctgcgacc acatgccgggtaaa (SEQ ID: No. 160) Con4-4 78 WYEQDACEWDPWTCEHMAEV tggtacgaacaggacgcttgcga atgggacccgtggacctgcgaac acatggctgaagtt (SEQ ID: 161¾ Con4-31 79 NRLQEVCEWDPWTCEHMENV aaccgtctgcaggaagtttgcgaa tgggacccgtggacctgcgaaca catggaaaacgtt (SEQ ID: No. 162) Con4 · C5 80 AATQEECEWDPWTCEHMPRS gctgctacccaggaagaatgcga atgggacccgtggacctgcgaac acatgccgcgttcc (SEQ ID: No. 163) Con4-42 81 LRHQEGCEWDPWTCEHMFDW ctgcgtcaccaggaaggttgcga atgggacccgtggacctgcgaac acatgttcgactgg (manufactured sequence: 164 No.) -30- 1317281 (26)

Con4-35 82 VPRQKDCEWDPWTCEHMYVG gttccgcgtcagaaagactgcga atgggacccgtggacctgcgaac acatgtacgttggt (SEQ ID: 165¾) Con4-43 83 SISHEECEWDPWTCEHMQVG tccatctcccacgaagaatgcgaa tgggacccgtggacctgcgaaca catgcaggttggt (SEQ ID: No. 360) Con4-49 84 WAAQEECEWDPWTCEHMGRM tgggctgctcaggaagaatgcga atgggatccgtggacttgcgaaca catgggtcgtatg (turn sequence: mm Con4-27 85 TWPQDKCEWDPWTCEHMGST acttggccgcaggacaaatgcga atgggatccgtggacttgcgaaca catgggttctact (SEQ Lake: No. 167) Con4-48 86 GHSQEECGWDPWTCEHMGTS ggtcactcccaggaagaatgcgg ttgggacccgtggacctgcgaac acatgggtacgtcc (SEQ ID: No. 168) Con4-46 87 QHWQEECEWDPWTCDHMPSK cagcactggcaggaagaatgcga atgggacccgtggacctgcgacc acatgccgtccaaa (SEQ ID: No. 169) Con4-41 88 NVRQEKCEWDPWTCEHMPVR aacgttcgtcaggaaaaatgcgaa tgggacccgtggacctgcgaaca catgccggttcgt (SEQ ID: 170 No.) Con4-36 89 KSGQVECNWDPWTCEHMPRN aaatccggtcaggttgaatgcaac tgggacccgtggacctgcgaaca catgccgcgtaac (sequence j: No. 171) - Con4-34 90 VKTQEHCDWDPWTCEHMR EW gttaaaacccaggaacactgcga ctgggacccgtggacctgcgaac acatgcgtgaatgg (_ sequence: No. 172) Con4-28 91 AWGQEGCDWDPWTCEHMLPM gcttggggtcaggaaggttgcga ctgggacccgtggacctgcgaac acatgctgccgatg (sequence Township J: 173¾) Con4-39 92 PVNQEDCEWDPWTCEHMPPM ccggttaaccaggaagactgcga atgggacccgtggacctgcgaac acatgccgccgatg (who do not sequence: No. 174)

•31- 1317281 _ (27) mMmm

Con4-25 93 RAPQEDCEWDPWTCAHMDIK cgtgctccgcaggaagactgcga atgggacccgtggacctgcgctc acatggacatcaaa (SEQ ID: No. Ί75) Con4-50 94 HGQNMECEWDPWTCEHMFRY cacggtcagaacatggaatgcga atgggacccgtggacctgcgaac acatgttccgttac (sequence Township J: No. Π6) Con4-38 95 PRLQEECVWDPWTCEHMPLR ccgcgtctgcaggaagaatgcgtt tgggacccgtggacctgcgaaca catgccgctgcgt (SEQ ID: No. 177) Con4-29 96 RTTQEKCEWDPWTCEHMESQ cgtaccacccaggaaaaatgcga atgggacccgtggacctgcgaac acatggaatcccag (SEQ ID: No. 178) Con4-47 97 QTSQEDCVWDPWTCDHMYSS cagacctcccaggaagactgcgtt tgggacccgtggacctgcgacca catggtttcctcc (J Township sequence: No. 179) Con4-20 98 QVIGRPCEWDPWTCEHLEGL caggttatcggtcgtccgtgcgaa tgggacccgtggacctgcgaaca cctggaaggtctg (SEQ ID: No. 180) Con4-45 99 WAQQEECAWDPWTCDHMVGL tgggctcagcaggaagaatgcgc ttgggacccgtggacctgcgacc acatggttggtctg (Sequence paste: (8)) Con4-37 100 LPGQEDCEWDPWTCEHMVRS ctgccgggtcaggaagactgcga atgggacccgtggacctgcgaac acatggttcgttcc (sequence: 182) · Con4-33 101 PMNQVECD WDPWTCEHMPRS ccgatgaaccaggttgaatgcga ctgggacccgtggacctgcgaac acatgccgcgttcc (SEQ ID: No. 183) AC2- Con4 102 FGWSHGCEWDPWTCEHMGST i ttcggttggtctcacggttgcgaat gggatccgtggacttgcgaacac atgggttctacc (SEQ ID: No. 184). Con4-32 103 KSTQDDCDWDPWTCEHMVGP aaatccacccaggacgactgcga ctgggacccgtggacctgcgaac acatggttggtccg Shu (SEQ identification Lee: 185) -32-1317281 __ (28) Description of the invention Continued ^

Con4-17 104 GPRISTCQWDPWTCEHMDQL ggtccgcgtatctccacctgccag tgggacccgtggacctgcgaaca catggaccagctg (ij of tender sequence: No. 186) Con4-8 105 STIGDMCEWDPWTCAHMQVD tccaccatcggtgacatgtgcgaa tgggacccgtggacctgcgctca catgcaggttgac (ij of Township sequence: No. 187) AC4- Con4 106 VLGGQGCEWDPWTCRLLQGW gttctgggtggtcagggttgcgaa tgggacccgtggacctgccgtctg ctgcagggttgg (SEQ ID: No. 188) Con4-l 107 VLGGQGCQWDPWTCSHLEDG gttctgggtggtcagggttgccag tgggacccgtggacctgctccca cctggaagacggt (ij of recognition sequences:? No. 189) Con4- C1 108 TTIGSMCEWDPWTCAHMQGG accaccatcggttccatgtgcgaa tgggacccgtggacctgcgctca catgcagggtggt (SEQ ID: No. 190) Con4-21 109 TKGKSVCQWDPWTCSHMQSG accaaaggtaaatccgtttgccag tgggacccgtggacctgctccca catgcagtccggt (SEQ ID: No. 191) Con4- C2 110 TTIGSMCQWDPWTCAHMQGG accaccatcggttccatgtgccag Tgggacccgtggacctgcgctca catgcagggtggt (sequence identification: No. 192) Con4-18 111 WVNEVVCEWDPWTCNHWDTP tgggttaacgaagttgtttgcgaat gggacccgtggacctgcaaccac tgggacaccccg (sequence identification: No. 193) Con4-19 112 WQV GMCQWDPWTCKHMRLQ gttgttcaggttggtatgtgccagt gggacccgtggacctgcaaacac atgcgtctgcag (sequence recognition ^ : No. 194) -33· 1317281

Con4-16 113 AVGSQTCEWDPWTCAHLVEV gctgttggttcccagacctgcgaat gggacccgtggacctgcgctcac ctggttgaagtt (turn sequence: No. 195) Con4-ll 114 QGMKMFCEWDPWTCAHIVYR cagggtatgaaaatgttctgcgaat gggacccgtggacctgcgctcac atcgtttaccgt (·· SEQ ID No. 196) Con4-'C4 115 TTIGSMCQWDPWTCEHMQGG accaccatcggttccatgtgccag tgggacccgtggacctgcgaaca catgcagggtggt (manufactured sequence: 197¾) Con4-23 116 TSQRVGCEWDPWTCQHLTYT acctcccagcgtgttggttgcgaat gggacccgtggacctgccagcac ctgacctacacc (ij of rinse sequence: No. 198) Con4-15 117 QWSWPPCEWDPWTCQTVWPS cagtggtcctggccgccgtgcga atgggacccgtggacctgccaga ccgtttggccgtcc (SEQ ID: No. 199) Con4-9 118 GTSPSFCQWDPWTCSHMVQG ggtacctccccgtccttctgccagt gggacccgtggacctgctcccac atggttcagggt (recognition sequence ^]: No. 200) TN8- Con4 4 QEECEWDPWTCEHM caggaagaatgcgaatgggaccc Atggacttgcgaacacatg (sequence identification: 201) LI-1 119 QNYKPLDELDATLYEHFIFHYT cagaactacaaaccgctggacga actggacgctaccctgtacgaaca cttcatcttccactacacc (sequence identification: 202) Ll-2 120 LNFTPLDELEQTLYEQWTLQQS Ctgaacttcaccccgctggacgaa ctggaacagaccctgtacgaaca gtggaccctgcagcagtcc (sequence identification · · 203) Ll-3 121 TKFNPLDELEQTLYEQWTLQHQ accaaattcaacccgctggacga actggaacagaccctgtacgaac agtggaccctgcagcaccag (sequence identification: 204Μ;) • 34- 1317281 _ (so) , ' <~ΙΙ·Ί · I wTim IH|M_<| _ULI>i

Ll-4 122 VKFKPLDALEQTLYEHWMFQQA gttaaattcaaaccgctggacgct ctggaacagaccctgtacgaaca ctggatgttccagcaggct (SEQ ID: No. 205) Ll-5 123 VKYKPLDELDEILYEQQTFQER gttaaatacaaaccgctggacgaa ctggacgaaatcctgtacgaacag cagaccttccaggaacgt (SEQ ID: No. 206) Ll-7 124 TNFMPMDDLEQRLYEQFILQQG accaacttcatgccgatggacgac ctggaacagcgtctgtacgaaca gttcatcctgcagcagggt (SEQ ID: No. 207) Ll-9 125 SKFKPLDELEQTLYEQWTLQHA tccaaattcaaaccgctggacgaa ctggaacagaccctgtacgaaca gtggaccctgcagcacgct (SEQ ID: No. 208) Ll-10 126 QKFQPLDELEQTLYEQFMLQQA cagaaattccagccgctggacga actggaacagaccctgtacgaac agttcatgctgcagcaggct i SEQ ID: No. 209) Ll-11 127 QNFKPMDELEDTLYKQFLFQHS cagaacttcaaaccgatggacga attggaagacaccctgtacaaaca gttcctgttccagcactcc (SEQ ID: No. 210) Ll-12 128 YKFTPLDDLEQTLYEQWTLQHV tacaaattcaccccgctggacgac ctggaacagaccctgtacgaaca gtggaccctgcagcacgtt (sequence Identification: No. 211) Ll-13 129 QEYEPLDELDETLYNQWMFHQR caggaatacgaaccgctggacga actggacgaaaccctgtacaacc agtggatgttdcaccagcgt (SEQ ID NO: 212) Ll-14 130 SNFMPLDELEQTLYEQFMLQHQ tccaacttcatgccgctggacgaa ctggaacagaccctgtacgaaca gttcatgctgcagcaccag (sequence identification: No. 213) Ll-15 131 QKYQPLDELDKTLYDQFMLQQG cagaaataccagccgctggacga actggacaaaaccctgtacgatca gttcatgctgcagcagggt (sequence identification: No. 214) -35- Description of the invention Continued page j 1317281 (31)

Ll-16 132 QKFQPLDELEETLYKQWTLQQR cagaaattccagccgctggacga actggaagaaaccctgtacaaac agtggaccctgcagcagcgt (SEQ ID: No. 215) Ll-17 133 VKYKPLDELDEWLYHQFTLHHQ gttaaatacaaaccgctggacgaa ctggacgaatggctgtaccacca gttcaccctgcaccaccag t (SEQ ID: No. 216) Ll-18 134 QKFMPLDELDEILYEQFMFQQS cagaaattcatgccgctggacgaa ctggacgaaatcctgtacgaacag ttcatgttccagcagtccc (SEQ ID: No. 217) Ll-19 135 QTFQPLDDLEEYLYEQWIRRYH cagaccttccagccgctggacga cctggaagaatacttgtacgaaca gtgeatccgtcgttaccac (SEQ ID: No. 218) LI-20 136 EDYMPLDALDAQLYEQFILLHG gaagactacatgccgctggacgc tctggacgctcagctgtacgaaca gttcatcctgctgcacggt (SEQ ID: No. 219) Ll-21 137 HTFQPLDELEETLYYQWLYDQL cacaccttccagccgctggacga actggaagaaaccctgtactacca gtggctgtacgaccagctg (·· SEQ ID No. 220) LI-22 138 YKFNPMDELEQTLYEEFLFQHA tacaaattcaacccgatggacgaa ctggaacagaccctgtacgaaga attcctgttccagcacgct (Sequence identification: No. 221) AC6-L1 139 TNYKPLDELDATLYEHWILQHS accaactacaaaccgctggacga actggacgctaccctgtacgaga ctggatcct gcagcactcc (SEQ ID: No. 222) Ll-Cl 140 QKFKPLDELEQTLYEQWTLQQR cagaaattcaaaccgctggacga actggaacagaccctgtacgaac agtggaccctgcagcagcgt (SEQ ID: No. 223) L1-C2 141 TKFQPLDELDQTLYEQWTLQQR accaaattccagccgctggacga actggaccagaccctgtacgaac agtggaccctgcagcagcgt (SEQ ID: No. 224) -36 · 1317281 (32) L1-C3 142 TNFQPLDELDQTLYEQWTLQQR accaacttccagccgctggacga actggaccagaccctgtacgaac agtggaccctgcagcagcgt (SEQ ID: No. 225) LI 6 KFNPLDELEETLYEQFTFQQ aaattcaacccgctggacgagctg gaagagactctgtacgaacagttt acttttcaacag (sequence bad Office J: No. 226) Conl-1 143 AGGMRPYDGMLGWPNYDVQA gctggtggtatgcgtccgtacgac ggtatgctgggttggccgaactac gacgttcaggct (sequence recognition ^: No. 227) Conl-2 144 QTWDDPCMHILGPVTWRRCI cagacttgggacgatccgtgcatg cacattctgggtccggttacttggc gtcgttgcatc i (sequence identification: No. 228) Coni-3 145 APGQRPYDGMLGWPTYQRIV gctccgggtcagcgtccgtacga cggtatgctgggttggccgaccta ccagcgtatcgtt (sequence): 229) Coni-4 146 SGQLRPCEEIFGCGTQNLAL tccggtcagctgcgtccgtgcgaa gaaatcttcggttg cggtacccag aacctggctctg (turn sequence: No. 230) Coni-5 147 FGDKRPLECMFGGPIQLCPR ttcggtgacaaacgtccgctggaa tgcatgttcggtggtccgatccag ctgtgcccgcgt (SEQ ID: 231¾) Con 1-6 148 GQDLRPCEDMFGCGTKDWYG ggtcaggacctgcgtccgtgcga agacatgttcggttgcggtaccaa agactggtacggt (SEQ ID: No. 232) 12-9-1 149 GFEYCDGMEDPFTFGCDKQT ggtttcgaatactgcgacggtatg gaagacccgttcaccttcggttgc gacaaacagacc ^ Sequence 渺J: No. 233) 12-9-2 150 KLEYCDGMEDPFTQGCDNQS aaactggaatactgcgacggtatg gaagacccgttcacccagggttg cgacaaccagtcc (sequence 渺j: 234) 12-9-3 151 LQEWCEGVEDPFTFGCEKQR ctgcaggaatggtgcgaaggtgtt -37- 1317281 (33) gaagacccgttcaccttcggttgc gaaaaacagcgt (sequence identification: No. 235) 12 -9-4 152 AQDYCEGMEDPFTFGCEMQK gctcaggactactgcgaaggtatg gaagacccgttcaccttcggttgc gaaatgcagaaa (sequence: No. 236) 12-9-5 153 LLDYCEGVQDPFTFGCENLD ctgctggactactgcgaaggtgtt caggaccc gttcaccttcggttgc gaaaacctggac (SEQ ID: 237¾) 12-9-6 154. HQEYCEGMEDPFTFGCEYQG caccaggaatactgcgaaggtat ggaagacccgttcaccttcggttg cga ataccagggt (SEQ ID: No. 238) 12-9-7 155 MLDYCEGMDDPFTFGCDKQM atgctggactactgcgaaggtatg gacgacccgttcaccttcggttgc gacaaacagatg (turn sequence: No. 239) 12-9-C2 156 LQDYCEGVEDPFTFGCENQR ctgcaggactactgcgaaggtgtt gaagacccgttcaccttcggttgc gaaaaccagcgt (sequence Miao J: 240¾) 12-9-C1 157 LQDYCEGVEDPFTFGCEKQR ctgcaggactactgcgaaggtgtt Gaagacccgttcaccttcggttgc gaaaaacagcgt (sequence identification: No. 241) 12-9 5 FDYCEGVEDPFTFGCDNH ttcgactactgcgaaggtgttgaa gacccgttcactttcggctgtgata accac (sequence identification: number 242)

In another embodiment, the invention relates to a performance vector comprising at least one polynucleotide of the present invention. In other specific embodiments, the invention relates to host cells comprising the expression vector. It is to be understood that the host cell is preferably a cell of a prokaryote (e.g., an E. coli cell) or a cell of a eukaryotic organism. The invention is also directed to pharmaceutical compositions comprising an effective amount of a composition as described herein in admixture with a carrier which is acceptable in Chinese. The present invention also relates to inhibiting unwanted angiogenesis in a mammal -38- 1317281

The method of <H (34)' includes administration of a therapeutically effective amount of a polypeptide or composition as described herein. The invention also relates to a method of modulating angiogenesis in a mammal' comprising administering a therapeutically effective amount of a polypeptide or combination as described herein. The invention more particularly relates to a method of inhibiting tumor growth characterized by unwanted angiogenic effects in a mammal' comprising administering a therapeutically effective amount of a polypeptide or composition as described herein. Further, the present invention is also directed to a method of treating cancer in a mammal comprising administering a therapeutically effective amount of a polypeptide or composition as described herein, and a chemotherapeutic agent. In a preferred embodiment the chemotherapeutic agent is at least one of 5; pu, CPT-11 and taxotere. However, it will be appreciated that other suitable chemotherapeutic agents and cancer therapeutic agents may also be used. The present invention also relates to methods of modulating at least one vascular permeability or plasma leakage in a mammal, including administration of a therapeutically effective amount. A polypeptide or composition as described herein. The invention further relates to the treatment of neovascular diseases, obesity, hemangioblastoma, hemangioma, atherosclerotic 'inflammatory disease, inflammatory disease, atherosclerosis, endometriosis in at least one eye in a mammal. Disease, neoplastic disease, bone-related disease or psoriasis' includes administering a therapeutically effective amount of a polypeptide or composition as described herein. It will be appreciated that the specific binding agents of the present invention can be used to treat a number of conditions associated with the elimination of unwanted or unwanted angiogenic effects. Such diseases include, but are not limited to, neovascularization of the eye, such as retinopathy (including diabetic retinopathy and age-related macular degeneration), psoriasis, blood vessels -39-1317281 (35) I silkworm mother cells Cancer, hemangioma, atherosclerosis, inflammatory disease 'like rheumatoid or lining: ruthenium h', sexual inflammatory disease, especially arthritis (including rheumatism, arthritis), $ tf * 4 His chronic inflammatory disease is like chronic asthma, arterial or post-transplant t-assisted atherosclerosis 'endometriosis and neoplasms, such as solid tumors and liquid tumors called Putian 2 i Like leukemia). For those who are familiar with this artist, f ^ " rumors can be seen by the additional disease treated with a single bond. Such additional diseases include, but are not limited to, obesity, vascular permeability*1*, Squeeze, leak, and bone-related diseases, including osteoporosis. Accordingly, the present invention is more directed to a method of treating such diseases associated with deregulated or unwanted production. <Other embodiments of the present invention will be more readily apparent from the disclosure provided herein. Figure 1 is a summary of the 'tumor volume (y-axis) pair in mice bearing A-431 tumors treated with the peptibody TN8-Con4-C of the present invention or phosphate buffered saline (PBS). The graph of time (X-axis) describes its details in the example. Figure 2 is a graph showing peptidic concentration (axis) versus time axis after administration in field mice treated with 5 x micrograms of 2xC〇n4 C, U 21n peptide. The details are described in the examples. Figure 3 depicts tumor volume (y-material) pairs in mice bearing A431 tumors treated with peptidom 2xC〇n4_c according to the invention, or phosphate buffered saline (PBS), or control peptibody. The graph of time (χ axis). The details are described in the examples. Figure 4 depicts the cultured A431 cells grown in vitro, represented by the peptibody c〇n4_c according to the present invention, the control peptibody-40-(36) (36) 13172281, or untreated. Figure. The details thereof are described in the examples. Fig. 5 illustrates the use of the peptone c〇n4_c, the peptibody LnN, the peptide LI 21-N, or the peptibody 2xcon4_c according to the present invention, or the physiological acid saline (PBS) buffered with a peak acid buffer. _Ang·2 antibody (Ab536) ' or Fc-treated c〇l〇205 tumor cells in the 'tumor volume (y_axis) versus time (χ_axis). The details are described in the examples. Figure 6 depicts tumors in mice bearing c〇i〇2〇5 xenograft tumors treated with various doses of the peptibody 2xCon4_c according to the invention, or phosphate buffered saline (1> guinea) or Fc. A plot of volume (y-axis) versus time (X-axis). The details are described in the examples. Figure 7 depicts tumor volume (y-axis) vs. time (X-axis) in mice bearing C〇l〇205 xenograft tumors treated with the peptibody 2xC〇n4_c according to the invention or the control peptibody. Diagram of the axis). Figure 7 also depicts a plot of cd3i staining area/total tumor area of these peptibodies. The details are described in the examples. Figure 8 depicts 'tumor in mice with c〇i〇2〇5 iso(iv) treated with peptibody 2xC〇n4_c according to the invention, or phosphate buffered saline (PBS), or control peptibody. Volume (4)) A plot of time (χ_axis). The details are described in the examples. The figure shows that the anti-Ang_2 body can inhibit the growth of C〇1〇205 tumors, regardless of when to start the drug. Figure 9 depicts a summary of the complete response (CR) rates obtained in antibody nude mice using either antibody Ab536 or peptibody 2xC〇n4_c in both A431 and Colo-205 xenograft models. The details are described in the examples. Figure 10A illustrates the use of a combination of peptibody 2x (: 〇n4_c, or -41 - 1317281 _ (37) 卜 ΙΜ 2xCon4-C according to the present invention, or phosphate buffered saline (PBS), Or a plot of tumor volume (y-axis) versus time (X-axis) in mice with C〇1〇205 xenograft tumors that are susceptible to PBS plus cancer. The details are described in the examples. Described with a combination of 2xCon4-C according to the invention, or a combination of 2xCon4-C and 5-FU, or phosphate buffered saline (PBS), or PBS plus 5-FU with C〇1〇 Tumor volume (y-axis) versus time (X-axis) in 205 xenograft tumord mice. Details are described in the examples. Figure 11A depicts the adjuvant-induced arthritis pattern in A graph of the degree of swelling of the foot (AUC soil SE) in the peptibody 2xCon4-C of the present invention, or phosphate buffered saline (PBS), or the control peptone, or the normal or arthritic control group. The details are described in the examples. Figure 11B depicts in the adjuvant-induced arthritis pattern, in the peptibo 2xCon4 according to the invention -C, or phosphate buffered saline (PBS), or a plot of mineral density (BMD) of the metacarpal bone in rats treated with control peptiforms, or normal or arthritic controls. Described in the examples Figure 11C depicts in the adjuvant-induced arthritis mode, in the peptibody 2xCon4-C according to the invention, or phosphate buffered saline (PBS), or as a control peptibody, or normal or arthritis A graph of changes in body weight in control-treated rats. Details are described in the examples. Figure 12 depicts two graphs depicting inhibition of VEGF-induced corneal angiogenesis in rats. The number of blood vessels measured in bovine serum albumin (BSA), VEGF plus phosphate buffered saline (PBS), or VEGF plus the peptidase Con4-C treated in the present invention. Second-42- 1317281 (38) The graphs depict the vessel area (square millimeters) in rats treated with BSA, VEGF plus phosphate buffered saline (pBS) or VEGF plus the peptibody Con4-C of the invention. Details are described in the examples. Figures 13A, 13B and 13C depict full length human Ang-2 (hAng-2), The N-terminal of hAng-2, and the C-terminus of hAng-2, the peptides TN8-Con4-C, L1-7-N and 12-9-3-C according to the present invention, and the control peptide The epitope mapping data (OD370) of the Tie2-Fc, C2B8 or 5B12. Details are described in the examples. Figure 14 depicts the binding affinity (KD) of the 2xCon-4-C peptibody according to the invention, using Sapidyne KinExA assay. The details are described in the examples. DETAILED DESCRIPTION OF THE INVENTION The paragraphs used herein are initially for organizational purposes only and are not to be construed as limiting the subject matter described in any way. Standard techniques can be used for the production of recombinant DNA molecules, proteins and antibodies, as well as tissue culture and cell transformation. Enzyme reaction and purification techniques, usually performed according to the manufacturer's instructions, or in the art 'using conventional procedures, as in Sambrook et al. (Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, Those stated in NY [1989]), or as described in this document. Unless specific definitions are provided, the terms used in connection with the laboratory procedures and analytical chemistry techniques, synthetic organic chemistry, and medical and pharmacological techniques described herein are those well known and commonly employed in the art. Standard techniques can be used for chemical synthesis, chemical analysis, pharmacological preparation, formulation and delivery, as well as patient treatment. -43· 1317281

Definitions The nouns used in this specification are defined as follows unless otherwise limited by specific examples.

Ang-2 — means a polypeptide (Tie-2 ligand_2) or a fragment thereof as set forth in Figure 6 of U.S. Patent No. 6,166,185, and related polypeptides, which are involved in the dual gene modification, Fusion variants, derivatives, substitutions, deletions and/or insertion variants, fusion peptides and polypeptides, and homologs between species. Ang-2 polypeptides may or may not contain additional terminal residues, such as leader sequences, The sequence, the amino terminal methionine, the amino terminal methionine and the lysine residue, and/or the tag or fusion protein sequence will depend on the manner in which it is prepared. " Biologically The term "active" when used in conjunction with an Ang_24 Ang_2 specific binding agent means that the peptide or polypeptide has at least one activity specific to Ang_2 or Ang-2 specific binding agent. A specific binding agent for Ang_2, possibly for at least one Ang- 2 biological activity, with agonist, antagonist, or neutralizing or blocking activity. "The term "specific binding agent" means a molecule, preferably a protein molecule 'specifically combined with Ang-2 , and its variants and derivatives, as in this article The specific binding agent may be a protein, a peptide, a nucleic acid, a hydrophobic compound, a lipid 'or a small molecular weight compound, which preferentially binds to Ang 2 . In a preferred embodiment, the specific binding agent according to the present invention is a peptide. Or a peptibosome, as well as a fragment, variant or derivative thereof, alone or in combination with other amino acid sequences, provided by known techniques. 5 Such techniques include, but are not limited to, enzyme cleavage, chemical incision, peptide synthesis or recombination. Technology. The invention of 1317281 _

(40) The MMMMM anti-Ang-2 specific binding agent is capable of binding to the binding moiety of Ang-2, regulating, for example, inhibiting or promoting the biological activity of Ang-2 and/or other Ang-2-related activities. The term "variant" as used herein includes a naturally occurring (or at least one known) amino acid sequence in which an amino acid residue is inserted into a binding agent, from which an amino acid residue is deleted. Bases, and/or those peptides and polypeptides substituted therein. The variants of the invention include fusion proteins as described below. "Derivatives" include those that have been chemically modified to modify in a manner different from that of inserting, deleting or replacing variants. "Specially combined with Ang-2", meaning the specificity of the present invention The ability of a binding agent (such as a peptibody or a peptide portion thereof) to recognize and interact with a mature, full-length or partial length human Ang-2 polypeptide or an ortholog thereof, such that, for example, Affinity ELISA or BIAcore assay as described herein to determine 'or neutralization capacity (by, for example, a neutralization ELISA assay as described herein, or a similar assay), for the same for any other blood vessel The pheromone, or the affinity or neutralization ability of other peptides or polypeptides, is at least 10 times as large, but may be as large as at least 5 ' as large as '100, 250 or 500 times, or even as large as at least 1 〇〇〇. , wherein the peptide portion of the peptibody is first partially fused to humans for evaluation in such assays. The term "antigenic epitope" means any molecule that is capable of An antigen-binding region of the binding agent 'are specific binding agents, such as peptides recognize a binding portion thereof and. The epitope is usually comprised of a surface group of a chemically active molecule such as a hydrazine such as an amino acid or a water-breaking compound side chain and having a specific -45· 1317281

Lai said Guan Gong! The structural characteristics of the three-dimensional space, as well as the specific charge characteristics. The epitopes used herein may be adjacent or not-adjacent. "Suppress and/or neutralize the epitope" as an epitope, when it is bound by a specific binding agent such as a peptide, the result is lost in vivo, in vitro or in situ. (or at least reduced) the biological activity of a molecule, cell or organism containing such an epitope. In the context of the present invention, the neutralizing epitope is located on or in association with the biologically active region of Ang-2. Alternatively, the term "activation epitope" is an epitope that, when combined with a specific binding agent of the invention, such as an antibody, activates, or at least maintains, a biologically active configuration of Ang-2. • • Peptide fragment " The term means a peptide or polypeptide that includes less than a complete, intact peptibodies. "The term "naturally occurring" when used with, for example, nucleic acid molecules, polypeptides, host cells, and the like When used together with biological substances, it means those found in nature and not modified by humans. • The word “separated”, when used with a specific binder of Ang-2 or Ang-2, By means of a compound which does not contain at least one contaminating polypeptide or compound found in its natural environment, and preferably is substantially free of any other contaminating mammalian polypeptide, which would interfere with its therapeutic or diagnostic use. When used with an Ang-2 peptibody or a fragment thereof, or a specific binding agent for Ang-2 of any other protein, means a peptide or polypeptide lacking a leader or signal sequence. For example, in a prokaryotic organism When a binding agent of the invention is expressed in a cell, the "mature" peptide or polypeptide may also contain additional amino acid residues (but still lacking a leader sequence), such as an amino terminal methionine-46-1317281 (42), or one or more methionine and lysine residues. It can be produced in this manner, with or without peptides or polypeptides that have removed these additional amino acid residues. •• Effective Content” or " therapeutically effective amount, when used in conjunction with Ang-2's specific binding agent, means the amount of that specific binding agent that is biologically active for one or more Ang-2 To the extent that supporting observable changes is useful or necessary. This change can be a degree of increasing or decreasing Ang-2 activity. This change is preferably to reduce the activity of Ang-2. The term "peptibody" is intended to include a molecule comprising an antibody Fc#*- site attached to at least one peptide. The production of a peptibody is generally described in PCT publication WO 00/24782, published May 4, 2000. The term ''variant' as used herein, includes those molecules such as peptide or peptide-media combinations, such as the peptibodies of the invention, wherein an amino acid residue is inserted into the amino acid of such a molecule. In the sequence, the amino acid residue is deleted from and/or substituted therein. The variant has one or more inserted amino acids, including fusion proteins as described below. ••Derivatives••including those A peptide and/or peptide-media combination, such as a peptide that has been chemically modified, modified in a manner different from the insertion, deletion or substitution of variants. The term "fragment" means a peptide or peptide-media combination, It includes less than the full length amino acid sequence of such peptide and/or peptide-vehicle combinations. Such fragments may result, for example, from truncation at the amino terminus, truncation at the carboxy-terminus, and/or deletion of internal residues from the amino acid sequence of the peptide or peptide-media combination. The fragment may be the result of another RNA ligation, and/or protease activity in vivo or in vitro. Can also be by chemical peptide synthesis, or by modifying the code •47, 1317281

Peptides, peptide-media combinations, or polynuclears of the Fc portion and/or peptide portion of the peptibody: y: acid, to construct such fragments. The term "Fc" means a type of vector of the invention and includes the sequence of a non-antigen-binding fragment of an antibody resulting from proteolytic digestion of the intact antibody, whether in the form of a monomer or a multimer. In the present invention, the source of Fc is preferably intact human Fc, and may be any immunoglobulin, although IgG1 and IgG2 are preferred. However, a Fc molecule derived from humans or from non-human species is also included herein. Fc's are composed of monomeric polypeptides which can be linked in the form of dimers or multimers by covalent (i.e., disulfide bonds) and non-covalent binding. Between monomeric subunits of a native Fc molecule, the number of intermolecular disulfide bonds ranges from 1 to 4, depending on the class (eg, IgG, IgA, IgE) or subclass (eg, IgG1, IgG2, IgG3, IgAl, IgA2) ) depending on. An example of a native Fc is a disulfide-bonded dimer resulting from papain digestion of IgG [see Ellison et al. (1982), >^(:1· Acids. Res. 10: 4071-9] As used herein, the term "natural Fc" is usually a monomeric, dimeric, and multimeric form of the form β "Fc functional site", including the natural Fc and Fc variant molecules, and As defined above, because of the Fc variant and the native Fc's, the term "Fc functional site" includes a monomeric or multimeric form of the molecule, whether digested from intact antibodies or produced by other methods. Systematic.

When applied to an Fc functional site or a molecule comprising an Fc functional site, the term "multimer" means having two or more covalent, non-covalent, or by covalent and non-covalent The interaction of the valence interaction with the molecular chain of the polypeptide chain e IgG molecule usually forms a dimer; IgM 'pentamer; IgD 'dimer; and IgA -48- invention Continuation: j 1317281 (44), monomer, two A polymer, trimer or tetramer. The multimer can be formed by utilizing the sequence of the native Ig source of Fc and the resulting activity, or by derivatizing (as defined below) the native Fc. When applied to a Fc functional site or a molecule comprising an Fc functional site, the term "dimer" means a molecule having two polypeptide chains joined in a covalent or non-covalent manner. The term "media" means a molecule that prevents degradation and/or increases half-life, reduces toxicity, reduces immunogenicity, or increases the biological activity of a therapeutic protein. Representative media include Fc functional sites, as well as linear polymers. (eg, polyethylene glycol (PEG), polylysine, dextran, etc.); a branched polymer (see, for example, Denkenwalter et al., U.S. Patent No. 4,289,872 issued Sep. 15, 1981; Tam, U.S. Patent No. 5,229,490, issued July 20, 1993; Frechet et al., WO 93/21259, issued Oct. 28, 1993; lipids; cholesterol groups (like steroids): carbohydrates or oligosaccharides Or any natural or synthetic protein, polypeptide or peptide that binds to a salvage receptor, as further described below. "Derivatives", "derived" or "derived", including the process and The resulting compound, wherein (1) a compound having a cyclic moiety: for example, cross-linking between cysteamine residues in the compound; (2) crosslinking the compound, or having a cross-linking position; The compound has a cysteamine sulfhydryl residue and thus forms a cross-linked dimer at the time of culture or in vivo; (3) substitution of one or more peptide bonds by a non-peptide bond: (4) -NRR1, -NRC(0)R1, -NRC(0)0R1, -NRS(0)2R1, -NHC(0)NHR, amber quinone imine group, or substituted or unsubstituted benzyloxy Carbonyl-NH-substitution-49-(45) (45) 13172281 茕埤 Description Continued use of N-terminal, wherein the ruthenium and R1 and ring substituents are as defined hereinafter: (5) by -C(0)R2 Or -NR3R4 replaces the C-terminus, wherein R2, R3 and R4 are as defined below, and (6) wherein the individual amino acid moieties are used throughout the treatment, and can be used with the selected side chain or terminal residue Base modified formulation modified compound. The derivatives are further described later. The term "peptide" means a molecule of from about 3 to about 75 amino acids, preferably about 5 to 50 amino acids, more preferably from 8 to 40 amino acids, and from about 10 to 25 Those of the amino acids are optimal. "Peptides can be naturally occurring or artificial (ie non-naturally occurring) amino acid sequences. Representative methods can be produced by any of the methods set forth herein. Peptides, such as those produced in peptide libraries (eg, phage display libraries), produced by chemical synthesis, derivatized by protein digestion, or produced using recombinant DNA technology. • 'Pharmacologically The term "active" means to determine the substance so described, having activity that affects medical parameters (eg, blood pressure, blood cell count 'cholesterol levels) or disease states (eg, cancer, autoimmune disorders, etc.). "antagonist peptide" or "inhibitor peptide" means the biological activity that blocks or interferes in some way with the protein of interest, or has a protein associated with the φ of interest. An antagonist or inhibitor that is comparable to a biologically active peptide. Thus the term 'ang-2-inhibitor peptide" includes a peptide that can be considered or derived as an Ang-2-antagonistic feature. Also included herein is a physiologically acceptable compound of the invention. Salts. ''Physiologically acceptable salts" means any specific examples of salts known or later found to be pharmaceutically acceptable: acetate; trifluoroacetate; argon halogenation , such as hydrochloride and argon bromide; -50-· 1317281 (46) sulfate; citrate; tartrate; glycolate: and oxalate, methylate and cantanate · peptide In one aspect, the invention relates to the development of Ang-2 peptibodies. The interaction of a protein ligand with its receptor usually occurs at a relatively large interface "however" as evidenced by the response to human growth hormones, There are only a few key residues at the interface that contribute most of the binding energy. Clackson et al., Science 267: 383-6 (1995). Large protein ligands display the binding epitopes in the correct-topology, or Provides Luenergy that is unrelated to the combination. So ' A molecule with a "peptide" length (usually 2 to 40 amino acids) binds to a receptor protein of a particular large protein ligand. Such peptides mimic the biological activity of large protein ligands ("peptides Agonists "), or inhibit the biological activity of large protein ligands via competitive binding ("peptide antagonists. 'The phage display technology has been visualized, when confirming such peptide agonists and antagonists' is effective Methods [See, for example, Scott et al, Science 249: 386 (1990); Devlin et al, Science 249: 404 (1990); June 29, 1993, issued US Patent No. 5,223, 4-9; 1998 US Patent No. 5, 733, 731 issued on March 31; I" US Patent No. 5, 498, 530 issued in March, I2, issued in March, 1995; US Patent No. 5,432,018 issued on July 11, 1995; U.S. Patent No. 5,338,665 issued on June 16th; U.S. Patent No. 5,922,545 issued July 13, 1999; W0 96/40987 published on December 19, 1996: and W0 published on April 16, 1998 98/15833 (incorporated herein by reference). In the display library, any peptide sequence can be displayed by fusion with the coat protein of filamentous phage, such as -51 - 1317281 (47) —|丨_,| _| - · I i'^l ^』 Preferably, the displayed peptide can be eluted with affinity to the extracellular functional site of the immobilized-antibody receptor. The retained phage can be enhanced by continuous cycling of affinity purification and repopulation. Peptides that bind optimally bind, and in one or more structurally related peptide families, key residues are identified. See, for example, Cwirla et al., Science 276: 1696-9 (1997), in which two different families are identified. Peptide sequences which can be safely displaced by alanine scanning or by mutation at the DNA level can also be proposed. The mutation library can be created and screened to more fully utilize the optimal binder. Lowman, Ann. Rev. Biophys. Biomoi. Struct. 26: 401-24 (1997). Peptides that mimic the binding activity of large protein ligands can also be proposed using structural analysis of protein-protein interactions. In this type of analysis, the crystalline structure can suggest the identity and related positions of the decisive residues of the large protein ligand from which the peptide can be designed. See, for example, Takasaki et al, Nature Biotech 15: 1266-70 (1997). These assays can also be used to investigate the interaction between the receptor protein and the peptide selected by phage display, which may suggest further modification of the peptide to increase binding affinity. Other methods compete with phage display in peptide studies. The peptide library can be fused to the carboxyl terminus of the lac repressor and expressed in E. coli. Other E. coli-based methods allow for display on the outer membrane of cells by fusion with peptidoglycan-bound lipoprotein (PAL). In the following, these and related methods are collectively referred to as "E. coli display". In other methods, translation of any RNA is stopped prior to ribosome release, resulting in the addition of its associated RNA to the polypeptide library. In the following, the method and related party -52-mmm 1317281 (10) are collectively referred to as "ribosome display." Other methods use chemical bonding of peptides to RNA. See, for example, Roberts and Szostak, Proc Natl Acad Sci USA , 94: 12297-303 (1997). Hereinafter, the method and related methods are collectively referred to as "RNA-peptide screening". A chemically derived peptide library has been developed in which the peptide is immobilized on a stable, non-biological material such as a polyethylene bar or a solvent permeable resin. Other chemically derived peptide libraries were scanned for peptides immobilized on slides using photolithography. In the following, it may be advantageous to collectively refer to this method and related methods as "chemical-peptide screening" beta chemical-peptide screening, which allows the use of D-amino acids and other non-natural analogs as well as non- - The components of the skin. Reviewing the biological and chemical methods in Wells and Lowman, Curr. Opin. Biotechnol., 3: 355-62 (1992). Conceptually 'any protein can be found using the sputum display and other methods mentioned above. Peptide mimetic. These methods have been used for mapping antigenic epitopes, confirming decisive amino acids in protein-protein interactions, and as a guide to discovering new therapeutic agents. See, for example

Cortese et al., Curr. Opin. Biotech. 7: 616-21 (1996). Peptide libraries are most often used in immunological research, such as epitope mapping. See

Kreeger, The Scientist 10(13): 19-20 (1996) 0 The peptides identified by screening the phage display library have been considered as "guide" rather than the therapeutic itself. Like other proteins and peptides, it may be rapidly filtered in the living body by the kidneys, by the cell clearance mechanism in the reticuloendothelial system, or by the degradation of proteolysis [Francis, (previous)] Except them. As a result, the art currently uses peptides, approves drug stems or as a scaffold for the design of organic compounds, which may not be as easy or as fast as confirmed by chemical library screening [53]. Lowman, (previously) 'Kay et al., (before)]» This technique will benefit from the process through which the therapeutic agent for anti-angiogenic effects can be more easily produced. Peptide junctions In a composition of a substance prepared according to the present invention, the peptide can be attached to the vehicle via the N-terminal or C-terminus of the peptide. Therefore, the medium-peptide molecules of the present invention can be traced by the following five polymers: (XOa-F^cx^, (Formula I) Xi-F, (Formula) f, -x2 (Formula III) Fi-(Li)c-Pi (Formula IV) Fi-(Li)〇-P1-(L2')h-P2 (Formula V) wherein:

Ft is the medium (preferably the Fc functional part); X2 is respectively selected from -(LA-P!, -, .(Ln_(L2)d-P2-(L3)e-P3 ' and -(LA-PHLJdULA-PHDf -P*, P! 'P2, P3 and P4 are respectively pharmaceutically active peptides as described herein; L2, L3 and L4 are respectively a linker; and "a","b", &quot ;c","d","e" and "f," respectively, or t, with the constraint that at least one of "a" and "b" is 1. Peptide The present invention is expected to selectively Binding or specifically binding to the peptide of Ang-2. Any number of such peptides, <soon, used in conjunction with the present invention. When producing peptides which can be used in the invention of the invention 54(50)(50)1317281 The phage display is a peptide ligand that shows the position of any position of the product that can be used from any of the sounds, because the affinity has been selected, because of the affinity of the peptide. D ~ He Ke 268: 23025-30 (1993) 〇 Man^ » J. Biol. Chem. The peptide of the present invention can be prepared by any method disclosed in the 蓺 _ _Technology. The use of a single-letter amine does not occur ^ ^ ^ Nest. Any "X" in the sequence (and in the present specification 'unless otherwise specifically embedded _. _ . 1 temple designation), means 20 naturally occurring (any of the amino acid residues. Or any non-naturally occurring amino acid that may be present (in the poems below), any of these peptides may be linked in tandem (ie, distant, ancient, Qilian α ground), with or without a linker, and for the column-joining case in the table. Use the "L μ column set tree to dip 2 4 to L to join the joint, and can be any of the descriptions described in this article For the sake of clarity, the column is separated by a dash and the linker is separated by a dash. Any peptide with a half-vehrobarine residue can be cross-linked with other Cys-containing peptides, any one or two. Any of these can be linked to a medium. Any peptide having more than one Cys residue can form a disulfide bond within the peptide. Any of these peptides can be derivatized as described herein with respect to which an amine group can be utilized. a derivative of a group-terminated terminal, and an amine group of the cap is -ΝΗ2. As for the amine group A derivative in which an acid residue is substituted with a moiety other than an amino acid residue, represented by S, is represented by Bhatnagar et al., J. Med. Chem. 39: 3814-9 (1996) and Cuthbertson et al., J. Any part described in Med. Chem. 40: 2876-82 (1997), which is incorporated herein by reference. All peptides are linked via a peptide bond to the β medium unless otherwise indicated. 55- 1317281 _ (8) 丨奁明 _5..-- ϋ In a specific embodiment, the present invention provides at least one peptide via the peptide (n) N-terminal, c-terminal or an amine-based acid residue The side chain is attached to at least one medium (F1, F2). Multiple vectors can also be used, such as Fc's' at each terminal or Fc at the terminal, and a PEG group at the other terminal or side chain.

The Fc functional site is a preferred medium. The & functional site can be fused to both the N or C terminal of the peptide, or the c-terminal. As mentioned above, within the scope of the invention, the Fc variant is suitable and glare can broadly modify the native Fc to form an Fc variant according to the invention, provided that the binding to the salvage receptor is still maintained. See, for example, W〇 97/34631 W0 96/32478. In such Fc variants, one or more of the native Fc can be removed: a position which provides an undesired structural or functional activity of the fusion molecule of the invention. For example, these positions can be removed by substituting or deleting residues 'insert residues at this position, or truncating the portion containing the position. The inserted or substituted residue may also be a modified amino acid such as a peptide mimetic or a D-amino acid. For many reasons, it may be desirable for Fc to change to H, as described below for several reasons. Representative & variants include molecules and sequences in which: 1. The position involved in the formation of disulfide bonds is removed. Such removal avoids the reaction with other cysteine-containing proteins in the host cells that are used to produce the molecules of the invention. For this purpose, fragments containing cysteine at the n• terminal may be truncated, or cysteine residues may be deleted, or substituted with other amino acids (e.g., propylamine, silk amine). Even when the cysteamine residue is removed, the single-chain Fc functional site can still form a dimer&> functional moiety' which is held together in a non-covalent manner. • 56· 1317281 (52) _ ait — 2. Modify the native Fc to be more compatible with the selected host cell. For example, a PA sequence close to the N-terminus of a typical native Fc can be removed, which can be recognized by digestive enzymes in E. coli, such as prolyl iminopeptidase. N-terminal thiosulphide residues can also be added, especially when the molecule is expressed recombinantly in bacterial cells such as E. coli. 3. Remove a portion of the N-terminus of the native Fc to prevent heterogeneity of the N-terminal when expressed in the selected host cell. For this purpose, any of the 20 amino acid residues preceding the N-terminus can be deleted, particularly those at positions 1, 2, 3, 4 and 5. 4. One or more glycosylation sites can be removed. Usually glycosylated residues (e.g., asparagine) can confer a lytic reaction. Such residues may be deleted or substituted with unglycosylated residues such as alanine. 5. Remove the location involved in the complement interaction, such as the Clq binding position. For example, the EKK sequence of human IgGl can be deleted or replaced. The recruitment of complement may not be advantageous for the molecules of the invention, and such Fc variants may be avoided. 6. Remove the location that affects binding to Fc receptors other than the salvage receptor. Natural Fc may have a position to interact with certain white blood cells, which is not required for the fusion molecules of the present invention, so it can be removed. 7. Remove the ADCC position. The ADCC position is known in the art. See, for example, Molec. Immunol. 29(5): 633-9 (1992) for ADCC positions in IgG1. This position is completely unnecessary for the fusion molecule of the present invention and can be removed. 8. When a native Fc is derived from a non-human antibody, the native Fc-57-Ferr page can be humanized. -.-«Jg 1317281 (53). Typically, in order to humanize a native Fc, the selected residue will be replaced in a non-human native Fc with a tether found normally in human native Fc. Techniques for humanizing antibodies are well known in the art. An alternative medium would be a protein, polypeptide, peptide, antibody, antibody fragment or small molecule (eg, a peptide mimetic compound) that is capable of binding to a salvage receptor. For example, the polypeptide described in U.S. Patent No. 5,739,277, issued to thess. Peptides can also be selected for binding to the FcRn salvage receptor by phage display. Such remedial receptor--binding compounds are also included within the meaning of "medium" and are within the scope of the invention. Such vectors should be selected for increasing half-life (e.g., by avoiding sequences recognized by proteases) and reducing immunogenicity (e.g., by preferring non-immunogenic sequences, as revealed in antibody humanization). As mentioned above, polymeric media can also be used with respect to Fi* F2. Various methods of attaching the chemical moiety that can be used as a medium are currently available, see, for example, the Patent Cooperation Treaty ("PCT" International Publication No. WO 96/11953, entitled "Chemical Modification of N-Terminal Proteins" N-Terminally Chemically Modified Protein Compositions and Methods", which is incorporated herein by reference in its entirety. A preferred polymeric medium is polyethylene glycol (PEG). The PEG group can have any convenient molecular weight and can be straight or branched. The average molecular weight of PEG will preferably be in the order of about 1,000 Torr. "kDa") is in the range of about 100 kDa, more preferably from about 5 kDa to about 50 kDa, preferably from about -58 to 1317281 - - (54) 5 kDa to about 10 kDa. The PEG group will typically be via a deuteration or reductive alkylation via a reactive group on the PEG moiety (eg, an aldehyde, amine, thiol or ester group) and reactivity on the compound of the invention. A group such as an aldehyde, amine or ester group is attached to a compound of the invention. A useful strategy for the PEGylation of synthetic peptides involves mixing the peptide and PEG moieties via a conjugated bond in solution, each carrying a particular functionality, and reacting to each other. Peptides can be readily prepared using conventional solid phase synthesis as known in the art. The peptide is pre-activated at the designated position using the appropriate functional group. The precursor is purified and fully characterized prior to reaction with the PEG moiety. The linkage of the peptide to PEG typically occurs in the aqueous phase. And can be easily monitored by reverse phase analysis HPLC. "The PEGylated peptide can be easily purified by preparative HPLC and characterized by analytical HPLC, amino acid analysis and laser desorption mass spectrometry." Polysaccharide polymer is Another type of water-soluble polymer that can be used for protein modification. Glucan is a polysaccharide polymer that includes individual subunits of glucose that are primarily linked by a 1_6 linkage. Many molecular weight ranges of glucan can be utilized' A fraction of from about 1 kDa to about 70 kDa can be readily utilized. For use in the present invention as a vehicle, the glucan itself or mixed with other media (e.g., Fc) is a suitable water soluble polymerization. See, for example, WO 96/1 1953 and WO 96/05309. The use of glucans conjugated to therapeutic or diagnostic immunoglobulins has been reported; see, for example, European Patent Publications No. 315,456, the disclosure of which is incorporated herein by reference in its entirety in its entirety in its entirety in the the the the the the the the the the the the The sheds - 1 joints can be linked to the "child" groups are arbitrary. When it appears, its chemical structure is not as good as it is because it is mainly used as a spacer. The linker is preferably composed of an amino acid, which is linked by a peptide bond. Therefore, in a better embodiment, In one embodiment, the linker consists of from 1 to 20 amino acids joined by a peptide bond wherein the amino acid is selected from the 20 naturally occurring amino acids. One or more of these amino acids can be glycosylated, as is well known to those skilled in the art. In a more preferred embodiment, the 1 to 20 amino acids may be selected from the group consisting of glycine, alanine, valine, aspartame, glutamine and amidic acid. More preferably, the linker is mostly composed of an amino acid having no steric hindrance, such as glycine and alanine. Thus, preferred linkers are polyglycines (especially (Gly) 5, (Gly) 8), poly (Gly-Ala) and polyalanine. Combinations of Gly and Ala are also preferred, such as those referred to herein as K1, and having the linker of the amino acid sequence set forth in the Examples herein. It is also possible to use non-peptide linkers. For example, an alkyl linker such as -NH-(CH2)s-C(0)-, wherein s = 2-20, can be used. These alkyl linkers may be further substituted by any non-sterically hindered group, such as a lower alkyl group (eg, a CrCd lower fluorenyl group, a halogen (eg, cn, Br), CN, NH2, phenyl, etc. A representative non-peptide linker is a PEG linker and has a molecular weight of from 100 kDa to 5000 kDa, preferably from 100 to 500 kDa. The peptide linker can be modified to form a derivative in the same manner as described above. Variants and derivatives of specific binding agents for bulk and derivative are also included within the scope of the invention 'including variants of insertions, deletions and substitutions in variants. It will be appreciated that the invention is in particular -60-1317281 (56) - .— - ·»» · ~~-ζ···α» Description of the invention continued Page 1 The specific binding agent may contain one, two or all three types of variants. The insertion and substitution variants may contain natural amines. a base acid, a conventional amino acid (as set forth below), or both. In one example, one or more amino acid residues, naturally occurring or unconventional amino acids, supplemental peptides are provided. Or an insertion variant of the amino acid sequence of the peptibody. The insertion can be in either or both of the protein Or may be located in the internal region of the peptidic amino acid sequence. Insertion variants with additional residues at either or both ends may include, for example, fusion proteins, as well as proteins comprising amino acid tags: ginseng or label. Inclusion variants comprising peptides and peptibodies, wherein one or more amino acid residues are added to the amino acid sequence of the peptide or peptidase, or a fragment thereof. The variant products of the invention also include mature peptides and Peptibodies in which the leader or signal sequence is removed and the resulting protein has an additional amine-based terminal sequence which may be natural or non-native. It is expected to have an additional oxime at position -1 of the amino acid. A specific binding agent for a thiamine thiol residue (such as a peptibosome) (Met·1-peptibody), such as an additional methionine and an lysine residue at positions -2 and -1 Specific binding agent (Met'Lys·1-). Variants with additional Met, Met-Lys, Lys residues (or one or more random residues, in general) for use in bacterial host cells It is particularly useful to increase the production of recombinant proteins. The invention also includes the use of specific performance resulting from the use of a specific binding agent for additional amino acid residues of the system. For example, using a portion of a glutathione-S-transferase (GST) fusion protein to express a desired vector of the desired polypeptide, such that After the GST component is cleaved in the polypeptide, the desired polypeptide carries additional glycine at the amino acid position-1. Variants that are also expressed in other -61 - 1317281 (57) vector systems are also contemplated, Included are those in which a poly-histidine tag is incorporated into an amino acid sequence, usually at the carboxyl and/or amine end of the sequence. Insert variants also include fusion proteins in which the amino group of the peptide or peptibody is made. And/or the carboxyl terminus is fused to other polypeptides, fragments thereof or amino acids and is generally not considered to be part of any particular protein sequence. Examples of such fusion proteins are immunogenic polypeptides, proteins with long circulating half-lives, such as immunoglobulin constant regions, labeled proteins, proteins or polypeptides that facilitate purification of the desired peptide or peptibosome, and promotion of A polypeptide sequence formed by a polymeric protein (such as a leucine zipper motif useful in dimer formation/stability). Insertion variants of this type typically have all or a substantial portion of the natural molecule linked to all or a portion of the second polypeptide at the N- or C-terminus. For example, fusion proteins typically use leader sequences from other species,

I to allow for the recombinant expression of the protein in a heterologous host. Other useful fusion proteins include the addition of immunologically active functional sites, such as the epitope of an antibody, to aid in the purification of the fusion protein. Including the incision site at or near the fusion junction will help remove the foreign polypeptide after purification. Other useful fusions include linking functional functionalities such as active sites derived from enzymes, glycosylation functional sites, cell targeting signals, or transmembrane regions. There are various commercially available fusion protein expression systems which can be used in the present invention. Particularly useful systems include, but are not limited to, glutathione-S-transferase (GST) system (Pharmacia), maltose-binding protein system (NEB, Beverley, -62- 励 1317281 (58) MA), FLAG system (IBI, New Haven, CT) and 6xHis system (Qiagen, Chatsworth, CA). These systems are capable of producing recombinant peptides and/or peptides that carry only a small amount of additional amino acid, which is unlikely to significantly affect the activity of the peptide or peptide. For example, both the FLAG system and the 6xHis system incorporate only short sequences, both of which are known to be poorly antigenic without adversely affecting the folding of the polypeptide into its native conformation. Another contemplated useful N-terminal fusion is the Met-Lys dipeptide fusion at the N-terminal region of the protein or peptide. Such fusions can produce a beneficial increase in protein performance or activity. # Other fusion systems produce peptide hybrids in which it is desired to excise the fusion partner from the desired peptide ® or peptide. In a specific embodiment, the fusion partner is linked to the recombinant peptoid by a peptide sequence containing a sequence that is specifically recognized by the protease. Examples of suitable sequences are those recognized by Tobacco Etch Virus Protease (Life Technologies, Gaithersburg, MD) or Factor Xa (New England B'iolabs, Beverley, MA). The invention also provides fusion polypeptides comprising all or a portion of a peptidomimetic or peptide of the invention, mixed with truncated tissue factor (tTF). tTF is a vascular targeting agent composed of a truncated form of human coagulation-inducing protein, which is a tumor vascular coagulant as described in US Patent Nos. 5,877,289: 6,004,555; 6,132,729; 6,132,730; No. 156,321; and the description of European Patent No. EP 0988056. Fusion of tTF with anti-Ang-2 skin or peptide' or a fragment thereof facilitates delivery of anti-Ang-2 to the stem cells. In another aspect, the invention provides deletion variants wherein one or more amino acid residues in a peptide or peptidic body are removed. Deletion can be accomplished at one or both ends of the peptide, or one or more residues within the peptidic amino acid sequence can be removed. Deletion variant -63- Hair surface fine 1317281 (59) It must include all fragments of the peptide or peptide. In another aspect, the invention provides substituted variants of the peptides and peptibodies of the invention. Substitutional variants include those in which one or more amino acid residues are removed and replaced with one or more additional amino acid residues. The amino acid may be naturally occurring or non-naturally occurring. of. Substituting the peptide or peptiboplast produced by the variant, "similar" the original peptide or peptidic body' has a certain percentage of amino acids in both molecules that are identical. Substituted variants in the peptide or peptide 'include 丨, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30 amino acid substitutions, the number of substitutions Up to 10% peptide or peptidic amino acid or more. In one aspect, the substitutions can be retentive in nature, however, the invention includes non-retentive substitutions as well as non-traditional amino acids. The identity and similarity of related peptides and peptibodies can be quickly calculated by known methods. Such methods include, but are not limited to, Computational Molecular Biology, Lesk, AM, ed., Oxford University Press, New York ( 1988); Biocomputing: Informatics and Genome Projects, Smith, DW, ed., Academic Press, New York (1993); Computer Analysis of Sequence Data, Part 1, Griffin, AM, and Griffin, HG Editor, Humana Press, New Jersey ( 1994); Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press (1987); Sequence Analysis Primer, Gribskov, M. and Devereux, J., ed., M. Stockton Press, New York (1991); and Carillo Et al., those described in SIAM J. Applied Math., 48: 1073 (1988). A preferred method for determining the relationship or identity of two peptides or polypeptides, or polypeptides and peptides, in order to provide the sequence of the test. The greatest match between the two. -64 - (4) Miscellaneous 1317281 (60) In a publicly available computer program, a method for determining identity is described. A preferred abandonment method for determining identity between two sequences, including but not limited to GCG suites, including GAP (Davereux et al, Nucl. Acid. Res., 12:387 (1984); Genetics Computer Group, University of Wisconsin, Madison, WI, BLASTP, BLASTN, and FASTA (Altschul et al, J. Mol. Biol 215:403-410 (1990)). From National Center for Biotechnology Information (NCBI) and other sources (BLAST Manual) Altschul et al., NCB/NLM/NIH Bethesda, MD 20894; Altschul et al., (previously (1990)) publicly available BLASTX programs. The well-known Smith Waterman algorithm can also be used to determine identity. The arrangement of amino acid sequences in a straight line arrangement results in pairing only short regions of two sequences, and this small alignment region may have extremely high sequence identity even if there is no sequence between the two full length sequences Significant relationship. Thus, in some embodiments, the selected alignment method (GAP program) will result in an arrangement that spans at least 10% of the full length of the target to be compared, ie, to compare at least 400 amine groups. a sequence of at least 40 consecutive amino acids, 30 consecutive amino acids at a sequence to be compared to at least 300 to about 400 amino acids, at least 200 to about 300 amine groups to be compared The acid sequence is at least 20 consecutive amino acids and is at least 10 consecutive amino acids at the sequence to be compared to about 100 to 200 amino acids. For example, using a computer algorithm GAP (Genetics Computer Group, University of Wisconsin, Madison, WI), aligns two polypeptides whose sequence identity is to be determined in order to match their respective amino acids -65-1317281 (61) (determined by algorithm) "Pairing spans"). In some embodiments, a gap open penalty point (which is typically calculated as a 3X average diagonal; the average diagonal) is used along with the algorithm to compare the diagonals of the matrix to be used. Average; "diagonal" is the fraction or number assigned to each optimal amino acid pair by a particular comparison matrix, and gap extension penalty (which is often 1/10 times the gap open penalty) ), and the comparison moment , Such as PAM 250 or BLOSUM 62. In some embodiments, the algorithm also uses a standard comparison matrix (for the PAM 250 comparison matrix, see Dayhoff et al., VIII 11 & 3 -Protein Sequence and Structure, 5(3) (1978); for BLOSUM 62 Comparison Matrix, see Henikoff et al, Proc. Natl. Acad. Sci USA, 89: 10915-10919 (1992). In some embodiments, the parameters of the polypeptide sequence comparison include the following: Algorithm: Needleman et al. J. Mol. Biol., 48: 443-453 (1970); Comparison Matrix: BLOSUM 62 from Henikoff et al., supra (1992); gap penalty: 12 gap length penalty: 4 threshold of similarity: 0. Using the above parameters, a GAP program may be useful. In some embodiments, the parameters mentioned above are preset parameters for comparison of polypeptides using the GAP algorithm (along with no penalty at the end gap) β In some embodiments, the parameters of the polynucleotide molecule sequence (as opposed to the amino acid sequence) are as follows: Algorithm: Needleman et al., supra (1970); comparison matrix: match = + 10, mismatch = 0 -66- 1317281(62) Clearance penalty: 50 gap length Degree penalty: 3 ^ Using the above parameters, the GAP program may also be useful. The parameters mentioned above are preset parameters for comparison of polynucleotide molecules. Other representative algorithms, gap opening penalty points, Gap extension penalty points, comparison moments, similarity thresholds, etc., are included in the Program Manual, Wisconsin Package, 9th Edition, September 1997. Specific choices will be familiar to the artist. Understand, and depending on the particular comparison to be made, such as DNA-to-DNA, protein-to-protein, protein-to-DNA: in addition, whether it is between specific sequence pairs (in this case) , GAP or BestFit is usually preferred, or a comparison between a sequence and a large sequence library (in this case FASTA or BLASTA is preferred). When used in this article, 20 traditional Amino acids and their vesicles are used routinely. See Immunology--A Synthesis (2nd ed., ES·Golub and DR Gren, ed., Sinauer Associates, Sunderland, Mass. (1991)) for any purpose Way of reference The herein. L or D amino acids may have the stereochemistry (except for Gly, which is also not not D L), and the polypeptides and compositions of the present invention may comprise a combination of stereochemistry. However, L stereochemistry is preferred. The present invention also provides opposite molecules in which the amino terminal termination to carboxyl terminal sequence of the amino acid is reversed. For example, the opposite of the molecule having the normal sequence Xi-Xs-X; will be X3-X2-X1. The present invention also provides an inverse-opposite molecule wherein, as above, the amino-terminal end-to-carboxyl terminal sequence of the amino acid is reversed and will normally be the "L" enantiomer-67-1317281 (63) I The residue of the inventor's political relics was changed to the form of the "D" stereoisomer. The stereoisomers of 20 conventional amino acids (for example, amino acids) and non-natural amines are like α- Γί ***. Α-glycol α-—11-substituted amino acid, Ν-alkylamino acid, lactic acid, and other non-greening compartments; the amine-free guanidine, which is a suitable group of the polypeptide of the present invention Examples of the 13⁄4 non-conventional amino acid include, but are not limited to, amino adipic acid, β-alanine, β-aminopropionic acid, aminobutyric acid '*hydropyridine acid, aminocaproic acid, amine group Heptanoic acid, aminoisobutyric acid, aminopimelic acid, diaminobutyric acid-locker-aminopimelic acid, diaminopropionic acid, N-ethylglycine, N-ethyl aspartate Indoleamine, hydroxy lysine, behenyl-hydroxy-amino acid, hydroxy guanamine, #------leucine, methyl-glycine, creatinine, N-methyl isoleucine, N•methyl-prolinic acid, n-leucine, and uranilide from 4-3⁄4-proline, 丫-carboxyglutamic acid, Bu AN, trimethyl lysine, ε-N-B柳其益吐*#· - TJ|.. 醒基离胺虹, 〇-phosphoric acid, Ν· 醯 丝 丝 丝, Ν-methyl methionine, 3-methylhistamine Acid, % by amino acid sulphonyl arginine, and other similar amino acids and amino acids (eg 4-hydroxyindole) Acid). Similarly, unless otherwise specified, the left-hand end of the single-stranded polynuclear acid sequence is the left-hand direction of the 5-terminal' double-stranded polynucleotide sequence, meaning the orientation. The 5, to 3, direction of the nascent RNA transcript is added, called the transcriptional direction; the sequence region on the DNA strand having the same sequence as the diterpene, and it is the 5' end of the 5' end of the RNA transcript. Is the "upstream sequence"; the sequence region on the DNA strand having the same sequence as the RNA, and it is 3, to the 3' end of the RNA transcript, it is called the "downstream sequence, ^ should be understood Based on its common side chain properties, the amino acid residues are divided into -68· 1317281 (64) \mmm class: L soil..., 'J raw dredging: alanine (Ala; A); Proline (val; v); leucine (Leu; L); isoleucine (lie; I); valine (Pr〇; p); tryptophan (Trp; W); phenylalanine ( Phe; F) and methionine (Met; M) 2. ±J±: glycine (Gly; G); serine (Ser; S); threonine (Thr; T); tyramine Acid (Tyr; Y); cysteine (CyS; C): amidoxime (Gin; Q); aspartic acid (Asn; N) and norleucine. 3. benign. Aminic acid (Asp; D), glutamic acid (Glu; Ε); 4. Age-related: lysine (Lys; K), arginine (Arg; R), histidine (His; Η) See Le Win, Β., Genes V, Oxford University Press (1994), p. 11. Reserving amino acid substitutions can include non-traditional amino acid residues, which are usually synthesized by chemical peptides rather than by Synthesis incorporation in biological systems. These include, but are not limited to, peptide mimics and other opposite or reverse forms of the amino acid moiety. Non-reserved substitutions may involve exchanging members of one of these species with those derived from other species. Members. In making such changes, according to certain embodiments, the hydrophobicity index of the amino acid can be considered. The hydrophobicity index of each amino acid has been assigned based on its hydrophobicity and charge characteristics. Isoleucine (+4.5); proline (+4.2); leucine (+3.8); albinoic acid (+2.8): cysteine/cystine (+2.5); methionine (+1.9); alanine (+1.8); glycine (-0.4); threonine (-0.7): serine (-0.8); tryptophan (-0.9); tyrosine (-1.3) ); proline (-1.6); histidine (-3.2); glutamic acid (-3.5); glutamine (-3.5); aspartic acid (-3.5); aspartame ( -3.5); lysine (-3.9); and arginine-69- (65) (65) 1317281 The invention states that platinum rhodium T'W — *fl (-4.5). The importance of hydrophobic amino acid indices in imparting biological functions to protein interactions is known in the art. Kyte et al., j. Mol. Bi〇l., 157:1〇5_13 1 (1982). It is known that certain amino acids can be substituted by other amino acids having a similar number or fraction of hydrophobic groups and still retain similar biological activities. When the change is based on a hydrophobic index, in some embodiments the 'substitution of the amino acid includes the amino acid beta having its hydrophobic index in the soil 2, in certain embodiments, including in the soil 1 Those within, and in some embodiments, those within ±0.5. In this context, it is also understood that the substitution of a similar amino acid can be carried out efficiently on the basis of hydrophilicity, in particular by creating a biologically functional peptide or which is intended to be used in a specific embodiment of immunology or Where the peptide is. In some embodiments, the maximum local average hydrophilicity of a protein, governed by the hydrophilicity of its adjacent amino acid, is related to its immunogenicity and antigenicity, i.e., the biological properties of the protein. These amino acid residues have been assigned the following hydrophilicity values: arginine (+3,0); lysine (+3.0); aspartic acid (+3.0±1); glutamic acid (+ 3.0±1),; serine (+0.3); aspartame (+0, 2); glutamine (+0.2); glycine (0); sulphate (-0.4): 脯Aminic acid (-0.5±1); alanine (-0.5); histidine (-0.5); cysteine (-1.0); methionine (-1.3); proline (-1.5); Leucine (-1.8); isoleucine (-1.8); tyrosine (-2.3); phenylalanine (-2.5) and tryptophan (-3.4). When modified on a similar hydrophilicity value, in some embodiments, the substitution of the amino acid includes an amino acid having a hydrophilicity value within ±2 'in certain embodiments' Within ±1 -70 - 1317281 (66) 丄 3 β 6 > 汐 those bricks of the invention, and in some embodiments, those within ± 〇. It is also possible to confirm the epitope of the antigen obtained from the original amino acid sequence based on the hydrophilicity. These regions are also referred to as "antigenic epitope core regions". Representative amino acid substitutions are set forth in Table 2 below. Table 2 Amino acid substitution The substitution represented by the original residue is preferably substituted for Ala Val, Leu, lie Val Arg Lys, Gin, Asn Lys Asn Gin, Glu, Asp Gin Asp Glu, Gin, Asn Glu Cys Ser, Ala Ser Gin Asn, Glu, Asp Asn Glu Asp, Gin, Asn Asp Gly Pro, Ala Ala His Asn, Gin, Lys, Arg Arg lie Leu, Val, Met, Ala, Phe, leucine Leu Leu leucine, lie , Val, Met, Ala, Phe lie Lys Arg, 1,4-diamino-butyric acid, Gln, Asn Arg Met Leu, Phe, lie Leu Phe Leu, Val, lie, Ala, Tyr Leu Pro Ala Gly Ser Thr , Ala, Cys Thr Thr Ser Ser Trp Tyr, Phe Tyr Tyr Trp, Phe, Thr, Ser Phe Val lie, Met, Leu, Phe, Ala, leucine Leu

-71 - 1317281 (67) Those skilled in the art will be able to determine appropriate variants of the polypeptides set forth herein using well-known techniques. In some embodiments, those skilled in the art can identify areas of the molecule that can be altered without disrupting activity by targeting areas that are not believed to be important for activity. In certain embodiments, residues and moieties in which a molecule is retained in a similar peptide or polypeptide can be identified. In certain embodiments, even regions that may be important for biological activity or structure may undergo undergoing retention amino acid substitution without disrupting biological activity or adversely affecting polypeptide structure. In addition, those skilled in the art can review structure-function studies to identify residues that are important for activity or structure in a similar polypeptide. From such comparisons, the importance of amino acid residues consistent with amino acid residues important for activity or structure in similar proteins can be expected in proteins. Those skilled in the art can choose a chemically similar amino acid substitution for such an amino acid residue that is expected to be important. Those skilled in the art can also analyze the relationship between the three-dimensional structure and the amino acid sequence, as well as structures in similar polypeptides. From this kind of information, those skilled in the art can predict the arrangement of the amino acid residues of the antibody and its three-dimensional structure. In certain embodiments, those skilled in the art may choose not to make radical changes to the amino acid residues expected to be on the surface of the protein, as such residues may involve important interactions with other molecules. Those skilled in the art can produce test variants which contain a single amino acid substitution at each desired amino acid residue. Variants can then be screened using activity assays known to those skilled in the art. Such variants can be used to gather information about appropriate variants. For example, if a change in a particular amino acid residue is found, it is destroyed -72- 1317281 (68) .- .10,··· Λ'< » <,,.* «-.m* - ~ -m ^ 'ill Λ"··, unwanted reduction, or inappropriate activity, can avoid such changes. In other words, based on information gathered from such routine experiments, those skilled in the art can readily determine that amino acids that are further substituted, either alone or in combination with other mutations, should be avoided. Many scientific publications have focused on the prediction of secondary structures. See Moult J., Curr. Op. in Biotech, 7(4): 422-427 (1996), Chou et al, Biochemistry, 13(2): 222-245 (1974); Chou et al, Biochemistry, 1 13(2): 21 1-222 (1974); Chou et al., Adv. Enzymol. Relat. Areas. Mol. Biol., 47:45-148 (1978); Chou et al., Ann. Rev. Biochem., 47:251-276 and Chou et al., Biophys. J., 26:367-384 (1979). In addition, the current use of computer programs to help predict secondary structure 3 - the method for predicting secondary structure is based on the same type of plasticity. For example, two polypeptides or proteins having more than 30% sequence identity, or more than 40% similarity, typically have similar structural topologies. The recent growth of the Protein Structure Database (PDB) has provided enhanced secondary structure predictability, including the number of possible folds in the structure of a polypeptide or protein. See Holm et al., Nucl. Acid. Res., 27(1): 244-247 (1999). It has been suggested (Brenner et al., Curr. Op. Struct. Biol) 7(3): 369-376 (1997)) that there are a limited number of folds in a particular polypeptide or protein, and once the critical number of structures has been resolved, Structural predictions become dramatic and more precise. Other methods of predicting secondary structure include "threading" (Jones, D., Curr. Opin. Struct. Biol., 7(3): 377-87 (1997); Sippl et al., Structure, 4(1): 15-19 (1996)), "Profile Analysis" (Bowie et al., Science, 253: 164-170 (1991); Gribskov et al., Meth Enzym·, 183:146-159 -73- 1317281 (69) (1990); Gribskov et al., Proc. Nat. Acad. Sci., 84(13): 4355-4358 (1987)) 'and" Evolution Chain " (see Holm, former (1999) and Brenner, former (1997)). In certain embodiments, a 'peptidomimetic variant includes a glycosylation variant in which one or more glycosylation sites, such as an N-linked glycosylation site, have been added to the peptidomimetic in. The N-linked glycosylation position is specific to the sequence: Asn-X-Ser or Asn-X-Thr, wherein the amino acid residue represented by X may be any amino acid residue other than valeric acid. . Substitution or addition of an amino acid residue creates the sequence' provides a new location where it is possible to add an N-linked carbohydrate chain. Or 'excluding the substitution of the sequence, removing the existing N-linked hydrophobic compound chain ^ also provides a rearrangement of the linked carbohydrate chains, excluding one or more small linkage glycosylation sites (usually those that occur naturally) and create one or more new Ν· connection locations. The present invention also provides "derivatives", which include peptibosome betas carrying different modifications, or additions, insertions, deletions or substitutions of amino acid residues, which modifications are preferably covalent in nature, and These include, for example, the combination of polymers, lipids, other organic and inorganic moieties. The gemstone, the scorpion, and the sputum sulphur can be used to prepare the derivative of the present invention, so as to increase the cycle of the body, or the quarantine period can be improved. The ability of the peptoid to aim at the desired cell, tissue or organ. & it lines one or more of the following modified representative derivatives including those already: have been bonded by non-belly-based [-ch2-〇c(〇)nr-]; phosphonate linkage Is -CH2-carbamate linkage-CH2-sulfonamide-74- 1317281 (70) Hairpin Description: Continued Poor: In addition, the side acid of the acid-removing amine anhydride reaction group, the package: :P is more than 2,4-pentyl-:[-CH2-S(〇)2NR-] linkage: urea [-NHC(0)NH-] linkage; -CH2-secondary amine linkage; or alkylation Peptidyl linkage [-C(0)NR6- 'where R6 is alkyl]' replaces one or more peptidyl [-C(0)NR-] linkages (bonds): wherein the N-terminal is derivatized -NRR1 group; -NRC(0)R group; -NRC(0)0R group; -NRS(0)2R group; -NHC(0)NHR group (wherein R and R1 are argon or low Carbon number alkyl, the restriction is that R and R1 are not both hydrogen; amber quinone imine group; 笮 oxycarbonyl · -NH-(CBZ-NH-) group; or 芊 oxycarbonyl _NH• a peptide having from 1 to 3 substituents on a phenyl group selected from the group consisting of a lower alkyl group, a lower number of oxo groups, a gas and a bromine group; wherein the free C terminal is derived as _c(〇)R2 (where R2 is selected Low-carbon group consisting of alkoxy) and _NR3R4 (wherein R3 and r4 respectively selected from the group consisting comprising and consisting of hydrogen lower alkyl) peptide. "Low carbon number" means a group having from 1 to 6 carbon atoms. It is also possible to use an amino acid residue that is targeted in the peptide and an organic derivatizing agent capable of reacting with a selected or terminal residue. The reaction, the individual amine groups are introduced into the polypeptide or composition of the invention. The following are typical: The sulfhydryl groups of the thiol and amine groups can be derivatized with succinate or other carboxylic acids L using these formulations, with the effect of reversing the residual weight of the base. Other suitable for the derivatization of residues containing "amino groups", sulphate trials of α + ^ 冢 methyl pyridazolium oxime ester; pyridinium phosphate, gas hydride, trinitrobenzene Continuation of acid; hydrazine methyl venous; - ketone' and reaction catalyzed by transaminase with glyoxylic acid -75- 1317281 (71) Modified spermine can be modified by reaction with one or several conventional reagents a mercapto residue, which includes benzamidine formaldehyde, 2,3-butanedione, 1,2-cyclohexanedione, and ninhydrin. Derivatization of arginine residues requires reaction under basic conditions. Because of the high pKa of the hydrazine functional group. In addition, these reagents can react with the amine group and the guanidine group of the arginine. The specific modification of the tyramine thiol residue itself has been extensively studied. Of particular interest is the introduction of spectral labels into the tyramine sulfhydryl residue by reaction with aromatic diazonium compounds or tetranitromethane. The most common is - N-acetylimidazole and four can be used. Nitromethane, which forms a 0-acetyl tyramine aryl group and a 3-derivative derivative, respectively, by means of a carbonized diimine (R'-N=C=N-R') Reaction, such as 1-cyclohexyl-3-(2-morpholinyl-(4-ethyl)carbodiimide or 1-ethyl-3-(4-azaindole-4,4-dimethylidene Carbonyl diimine, selectively modifying a carboxyl side chain group (aspartame or amidoxime). In addition, by reacting with ammonium ions, aspartame and valley The amine sulfhydryl residue is converted to anthraquinone oxime and imide amine sulfonyl residue. The glutamine amidoxime and the aspartame sulfhydryl residue are usually deaminated to a corresponding The residues of the amine and the aspartame can be re-aminated under weakly acidic conditions. Any of these residues are within the scope of the invention. The derivatization of the preparation is useful for crosslinking the peptide or its functional derivative with a water-insoluble support matrix or other macromolecular carrier. Commonly used linkages include, for example, 1,1-double (heavy atmosphere) 2-phenylethane, glutaraldehyde, N-hydroxysuccinimide, for example, ester with 4-azide-76-1317281 (72) salicylic acid, high dual function quinone Ester' includes two glass shots Imino esters, such as 3,3·-dithiobis(succinimide capric vinegar) and dual-functional maleimide, such as bis-indole-cisene Amino-1'8-octane. A derivatizing agent such as methyl-3-[(p-azidophenyl)dithio] propylene acetal produces a photoactivatable intermediate which is capable of The presence of light forms crosslinks. Alternatively, a reactive water-insoluble matrix can be used for 'fixing proteins', as in U.S. Patent Nos. 3,969,287; 3,691'〇16; 4,195,128: 4,247,642; 4,229,537 and 4,330,440. No.: The hydrocyanide-activated hydrophobic compound and reactive substrate. Other possible modifications include the phosphorylation of proline and the hydroxylation of the lysine, the phosphorylation of the hydroxyl group of the serine or sulfhydryl residue, the oxidation of the sulfur atom in the Cys, the acid, Methylation of α-amino groups of arginine and histidine side chains (Creighton, TE·, Proteins: Structure aiid Molecule Properties, WH Freeman & Co., San Francisco, pp. 79-86 ( 1983)) 'Ethylation of N-terminal amines, and in some cases *A-terminal carboxyl group amide amidation. Such derived moieties preferably improve one or more of the characteristics, including the anti-angiogenic activity, solubility, absorption 'biological half-life of the compound, and the like. Alternatively, the derivatized moiety can have the same or substantially the same characteristics and/or characteristics as the underived compound. This moiety may additionally exclude or reduce any unwanted side effects of the compound, and the like. In addition, any part of the DNA sequence of the present invention can be changed at the DNA level to be changed to a further compound. The compound can be combined with the selected host cell-77-(73)(73)1317281 codon 1 in Escherichia coli, k > The code is known for this technique Λ β , , , , ^ 肀. The codon can be substituted to exclude the restriction position, or include a restriction position that is inactive, which can facilitate the processing of DNA in the selected king cells.

庋*丨,# and 6 α tamper with the media, linker and peptide DNA sequences to include any life +, λα — 士... 序列 sequence changes. Therefore, all the tampering and substitution in this article, saying that the female "life role" is equally applicable to all the defects of the present invention, including 仞^人. The kernel is not limited to peptides, peptide dimers and many Polymers, Linkers, and Vectors In addition, those who are familiar with this art can review the structure-function studies to identify residues that are important for activity or structure in a similar form. From this comparison, it can be expected in the peptide. The importance of an amino-based residue amino acid S-acid residue that is important for activity or structure in a similar peptide. Those skilled in the art can choose to chemically select the amino acid residue of such a peptide that is expected to be important. Similar amino acid substitutions. Those skilled in the art can also analyze the three-dimensional structure and the amino acid sequence, and the relationship with the structure in a similar polypeptide. From this information, the amine of the peptide can be expected. The arrangement of the base acid residues and their three-dimensional structure. Those skilled in the art may choose to base the amino acid residues expected on the surface of the protein without radical changes, as such residues may be involved with other molecules. The important interaction. In addition, those skilled in the art can produce test variants which contain a single amino acid substitution at each desired amino acid residue. The activity assay known to the artist can then be used to screen for changes. Such variants can be used to collect information about appropriate variants. For example, if a change in a particular amino acid residue is found, the damage is undesirably reduced, or uncomfortable -78- 1317281 (74) By inventing the activity, it is possible to avoid such changes. In other words, based on the information collected from such routine experiments, those skilled in the art can easily determine that they should be avoided alone or in combination with other mutations to further replace them. Amino acids. Many scientific publications have been devoted to the prediction of secondary structures.

Moult J., Curr. Op. in Biotech” 7(4): 422-427 (1996), Chou et al.

Biochemistry, 13(2): 222-245 (1974); Chou et al, Biochemistry, 1 13(2): 21 1-222 (1974); Chou et al., Adv. Enzymol. Relat. Areas

Mol. Biol., 47:45-148 (1978); Chou et al, Ann. Rev. Biochem., - 47:251-276 and Chou et al, Biophys. J., 26:367-384 (1979). In addition, computer programs are currently available to help predict secondary structure. One method of predicting secondary structure is based on isomorphic modeling. For example, two polypeptides or proteins having more than 30% sequence identity, or more than 40% similarity, typically have similar structural topologies. The recent growth of the Protein Structure Database (PDB) has provided enhanced secondary structure predictability, including the number of possible folds in the structure of a polypeptide or protein. See Holm et al., Nucl. Acid. Res., 27(1): 244-247 (1999). It has been suggested (Brenner et al., Curr_ Op. struct. Biol., 7(3): 369-376 (1997)) that there are a limited number of folds in a particular polypeptide or protein, and once the structural number of structures has been resolved' The prediction of the structure becomes dramatic and more precise. Other methods for predicting secondary structure include "Threading" (Jones, D., Curr. 0pin. Struct. 7(3): 377-87 (1997); Sippl et al., Stmcture, 4(1): 15-9 ( 1996)), "Profile Analysis" (Bowie et al., Science, 253: 164-170 (1991); Gnbskov et al., Meth Enzym, 183: 146_159 (199 〇); Gribsk〇v et al., Proc. Nat Acad. Sci., 84(13): 4355-8 (1987)), and "Evolution Chain" -79- 1317281 (75) (see Holm, formerly and Brenner, prior. The invention also includes derivatives The specific binding agent 'e.g., peptibody' is covalently modified to include one or more water soluble polymer attachments, such as polyethylene glycol, polyoxygen& diol, or polypropylene glycol, as in U.S. Patent No. 4,640,835; 4 496, 689; 4, 301, 144; 4, 670, 417; 4, 791, 192 and 4, 179, 337. Other useful polymers are known in the art, including monomethoxy-polyethylene glycol, chlorinated, Cellulose or other polymer based on water-breaking compounds, poly-(N-ethyl-associated phloprofenone)-polyethylene glycol, propylene glycol-alloy homopolymer, polyglycolide/ Oxygenated co-polymers, polyoxyethylated polyols (such as glycerol) and polyethylol, and mixtures of these polymers are particularly preferred for covalent modification with polyethylene glycol (PEG) subunits. Peptide. It can be bound to a water-soluble polymer at a specific position, for example, at the amino terminus of the peptibody, or randomly attached to one or more side chains of the polypeptide. β uses PEG to improve the specific binding agent. For example, the therapeutic efficacy of the peptibody, and the humanization of the antibody, as described in, for example, U.S. Patent No. 6,133,426 issued to Gonzales et al., issued Jan. 17, 2000. Or a mediator. Such derivatives may improve the solubility, absorption, biological half-life of the compound, and the like. This moiety may additionally exclude or reduce any unwanted side effects of the compound, and the like. Representative derivatives Included are compounds wherein: 1. The compound or some portion thereof is cyclic. For example, a 'modifiable peptide moiety' may contain two or more Cys residues (eg, as a linker), which may be disulfide-bonded form Cyclize it. 2. The compound crosslinks between molecules or makes it crosslinkable. -80· (76) (76) 13172281 For example, 'the peptide moiety can be modified to contain a Cys residue, and By virtue of the ability to form intermolecular disulfide bonds with similar molecules, the compounds can also be crosslinked via their C-terminus. 3. Replace one or more peptidyl [-C(0)NR-] linkages (bonds) by non-peptidyl linkages. A representative non-peptidyl linkage is -CH2-carbamate [-CH2-0C(0)NR-], phosphonate, -CH2-sulfonamide [-CH2-S(0)2NR- ], urea [-NHC(0)NH-], -CH2-subamine, and alkylated peptide [_c(〇)NR6-, wherein R6 is a lower alkyl group]. 4. Deriving the N-terminal. Typically, the N-terminus can be thiolated or modified to a substituted amine. Representative N-terminal derived bases include -NRRi (other than -NH2), -NRC^O)!^, -NRC(0)0R, -NRS(0)2R, -NHC^NHR!, amber An imine, or benzyloxycarbonyl-NH-(CBZ-NH-), wherein the sizing and the heart are respectively argon or a lower alkyl group, and wherein the phenyl ring may be one to three selected from the group consisting of Ct-C4, The substituent of the group consisting of C1-C4-precipitated oxy-chloro and bromo substituted β. 5. Derived from a free C-terminus. Typically, the C-terminus can be esterified or decylated. For example, (NH-CH2-CH2-NH2)2 can be added to the compound of the invention at the C-terminus using the methods described in the art. Similarly, -NH2 can be added to the compounds of the invention at the C-terminus using the methods described in the art. Representative C-terminal derivatizing groups include, for example, -C(0)R2, wherein ulmen 2 is a lower alkoxy group, or -NR3R4, wherein 113 and r4 are each hydrogen or CrC8 alkyl (preferably Ci) -G alkyl). 6. Replace the disulfide bond with another, preferably more stable, cross-linking moiety (eg, a stretching base). See, for example, Bhatnagar (formerly); Alberts et al. Thirteenth Am. Pep. Symp., 357-9 (1993) 0 • 81 - 1317281 (77)

DESCRIPTION OF THE INVENTION continued .:--«-r , i.i . V , -U 7. Modification of one or more individual amino acid residues. Derivatives which specifically react with selected side chains or terminal residues are known, as will be described in detail below.

Residues from the amine sulfhydryl residue and the amine terminal may be reacted with succinic acid or other carboxylic anhydride to reverse the charge from the amine sulfhydryl residue. Other preparations suitable for deriving residues containing an α-amino group, including quinone imide, such as methyl pyridinium imide; pyridoxal phosphate; pyridoxal; gas borohydride; trinitrobenzene sulfonate Acid; 0-methylisourea; 2,4-pentanedione; and a reaction catalyzed by a transaminase of glyoxylic acid. The spermine sulfhydryl residue can be modified by reaction with any one or several conventional reagents, including benzamidine formaldehyde, 2,3-butanedione, 1,2-cyclohexanedione, and ruthenium. ketone. Derivatization of arginine residues requires reaction under basic conditions due to the high pKa of the oxime functional groups. In addition, these reagents can react with the base groups of the amine acid and the ε-amino group of the arginine.

Specific modifications of tyramine sulfhydryl residues have been extensively studied, and it is of particular interest to introduce spectral markers into tyramine thiol residues by reaction with aromatic diazonium compounds or tetranitromethane. . Most commonly, quinone-ethenyl imidazole and tetranitromethane can be used to form a 0-ethyl decyl guanamine species and a 3-nitro derivative, respectively. By reaction with carbodiimide (R'-N=C=N-R'), such as 1-cyclohexyl-3-(2-morpholinyl-(4-ethyl)carbodiimide Imine or 1-ethyl-3-(4-azaindole-4,4.-dimethylidene)carbodiimide, selectively modifying carboxyl side chain groups (aspartame or glutamine) In addition, by reacting with ammonium ions, the aspartame and glutamine residues can be converted to aspartamide-82- 1317281 (78) group and glutamine amidoxime. The residue can be deaminated and aminated to a corresponding glutamine and aspartic acid residue. Alternatively, it can be weakly acidic. These residues are deaminated under conditions. Any of these residues are within the scope of the present invention. The cysteamine-based residue may be replaced by an amino acid residue or other moiety to exclude a sulfur bond. Knot 'or conversely stabilize cross-linking β see, for example, Bhatnagar (previously). Derivatization of a dual-functional formulation for the exchange of a peptide or a functional derivative thereof with a water-insoluble support matrix or other macromolecular carrier Union is useful. Often Linkers include, for example, 1,1-bis(diazonium)-2-phenylethane, glutaraldehyde, N-hydroxysuccinimide, such as esters with 4-azidosalicylic acid Highly functional sulfhydryl esters, including disuccinimide esters, such as 3,3·-dithiobis(succinimide propionate), and dual-functional maleicene An amine such as bis-N-maleimidoimino-1,8-octane oxime such as methyl-3-[(p-azidophenyl)dithio]propanimide or the like a derivatizing agent that produces a photoactivatable intermediate capable of forming crosslinks in the presence of light. Alternatively, in order to immobilize a protein, a reactive water-insoluble matrix can be used, as in U.S. Patent No. 3,969,287; , 016; 4, 195, 128; 4, 247, 642; 4, 229, 537 and 4, 330, 440, the sulphuric acid activated breaking water compound and reactive substrate. The carbohydrate (oligosaccharide) can be conveniently attached It is known in the protein to be at the position of the glycosylation site. Usually, the 〇-linked oligosaccharide is attached to a serine (Ser) or threonine (Thr) residue. 'At the same time the N-linked vinegar-83- 1317281 (79) Illustrated continuation sheet j attached to the asparagine (Asn) residue, at which point they are part of the sequence Asn-X-Ser/Thr Wherein X may be any amino acid other than proline. X is preferably one of the 19 naturally occurring amino acids other than proline. The structure of N-linked and 0-linked oligosaccharides, and The types of sugar residues found in the various types are different. The type of sugar often found in both is N-acetyl-neuraminic acid (called sialic acid). Sialic acid is often N-linked and The terminal residue of the 0-linked oligosaccharide, and by virtue of its negative charge, imparts acidic properties to the glycosylated compound. Such positions may be incorporated into: a linker of a compound of the invention, and preferably glycosylated by a cell during recombinant production of the polypeptide compound (e.g., in a mammalian cell, such as CH0, BHK, C0S). However, such positions can be glycosylated by synthetic or semi-synthetic procedures known in the art. Other possible modifications include hydroxylation of proline and lysine, phosphorylation of the hydroxyl group of the serine or sulfhydryl residue, oxidation of the sulfur atom in Cys, lysine , methylation of α-amino groups of arginine and histidine side chains [Creighton, Proteins: Structure and Molecular Properties (WH Freeman & Co·, San Francisco), pp. 79-86 (1983) )].

Alternatively, the compounds of the invention may be altered at the DNA level. The DNA sequence of any portion of the compound can be altered to a codon that is more compatible with the host cell of choice. With regard to E. coli, which is the best host cell, the most suitable codons are known in the art. Codons can be substituted to exclude restricted positions, or include inactive restricted positions, which can aid in the processing of DNA in selected host cells. The modifiable medium 'linker and peptide DNA-84-(80) (80) 13172281 爹.a month says that the sequence 'includes any of the aforementioned sequence changes.觐 and maturity - A specific embodiment of the invention includes "affinity mature"" peptides and peptides. This process attempts to increase the affinity or bio-activity of the peptides and peptibodies of the present invention using phage display or other selection techniques. Based on the _ sequence (produced for the collection of related peptides), the sputum production refers to the secondary phage display library of 丨, where the "core, amino acid (determined from the consensus sequence) is invariably retained Or tend to occur frequently. Alternatively, individual peptide sequences can be used to produce a phage display library that is biased towards one side, and the phage display library can be used to improve the binding of Ang-2, or Peptides with enhanced biological activity (which can be converted into peptibodies). Non-peptide analogs/peptone mimetics In addition, non-peptide analogs of peptides are also expected which provide stabilized structures for reduced biodegradation Based on the selected inhibitory peptide, a peptide mimetic analog is prepared by replacing one or more residues with a non-peptide moiety, and the non-peptide portion preferably allows the peptide to retain its natural confirmation, or is stable and better. For example, it has been confirmed by biological activity that it retains the ability to recognize and bind to Ang-2. In one aspect, the resulting analog/mimetic exhibits increased binding affinity for Ang-2. In Nachman et al., Regul. Pept 57:359-370 (199 In 5), an example of a method for preparing a non-peptide mimetic analog from a peptide is described. β If desired, the peptide of the present invention can be modified, for example, by glycosylation, guanylation, carboxylation or phosphoric acid. By chemical action, or by the acid addition salt, the amine, the vinegar, especially the C-terminal vinegar, and the hydrazine-hydrazine derivative of the peptide of the present invention. The peptide body can also be modified to form with other parts. Covalent -85 - 1317281 (81) Description of the Invention Continued Page 3 or #covalent complex' to create a peptide derivative by virtue of the chemical moiety and the amino acid side chain including the peptibody, or N- Or the cross-linking of functional groups at the C-terminus to prepare a covalently-bound complex beta, specifically looking for a peptide conjugated to a reporter group, including but not limited to radioactive labels, fluorescent labels An enzyme (eg, a catalytic colorimetric or fluorescent reaction), a substrate, a solid phase matrix, or a carrier (eg, biotin or avidin). The invention provides a molecule comprising a peptidomimetic molecule, wherein the molecule preferably includes The reporter group 'selected from radioactive labels, fluorescent labels, A group of enzymes, substrates, solid phase matrices, and carriers. Such labels are well known to those skilled in the art, such as biotin labeling, which is particularly desirable. The use of such labels is well known to those skilled in the art, and Other useful labels include, but are not limited to, radioactive, fluorescent, and chemiluminescent labels, for example, in U.S. Patent Nos. 3,817,837; 3,850,752; 3,996,345 and 4,277,437. U.S. Patent Nos. 3,817,837; 3,850,752: 3,939,350 and 3,996,345. Any of the peptibodies of the present invention may comprise one, two or more of any of these labels. The method of making the peptides can be used in a variety of ways. The techniques known in the art to produce the peptides β of the present invention, for example, can be synthesized according to conventional techniques, in solution, or on a solid support. There are various commercially available automatic synthesizers that can be used according to known drafts. See, for example, Stewart and Young (previously);

Tam et al, J. Am. Chem. Soc., 105:6442 (1983); Merrifield, Science 232:341-347 (1986); Barany and Merrifield, The Peptides. Gross and 1317281 (82)

faMU

Edited by Meienhofer, Academic Press, New York, 1-284; Barany et al, Int. J. Pep. Protein Res, 30: 705-739 (1987); and U.S. Patent No. 5,424,398, each incorporated by reference In this article. The solid phase peptide synthesis method uses a copolymer (styrene-divinylbenzene) containing 0.1 to 1.0 mM of amine per gram of polymer. These methods for synthesizing peptides use an ethoxycarbonyl (t-BOC) or 9-fluorenylmethoxycarbonyl (FMOC) protected a-amino group. Both methods involve stepwise synthesis whereby in each step a single amino acid is added starting from the C-terminus of the peptide (see, Coligan et al., 〇111·!·.?!^.-Immunol., Wiley. Interscience, 1991, unit 9). Upon completion of the chemical synthesis, the synthetic peptide can be deprotected to remove the t-BOC or FMOC amino acid blocking group' and cleaved from the polymer by treatment with an acid at a reduced temperature (eg, liquid HF-10% anisole, about 0.25 to about 1 hour at 〇 ° C). After evaporating the reagent, the peptide is extracted from the polymer using a 1% acetic acid solution and then freeze-dried to yield a crude material. It can be condensed by using 5% acetic acid as a solvent, such as on the cross-linked dextran G-15. Techniques such as gel filtration 'normally purify the crude material. Freeze-drying of the appropriate fraction of the column will result in a homogeneous peptide or peptide derivative which can then be analyzed by, for example, amino acid analysis, thin layer chromatography, high performance liquid chromatography, ultraviolet absorption spectroscopy, moirization Standard techniques such as solubility, characterizing, and quantifying β by solid phase Edman degradation can also utilize other methods, such as selecting peptides from a phage display library, from amino acids as described herein. Group, to prepare the library. Phage display may not be particularly effective in identifying peptides that can be used in accordance with the present invention. The preparation of the sputum cell library (using, for example, ml 13 or λ phage), shows an insert from 4 to about 80 amino acid residues. The insert can represent, for example, an array that is completely degraded or favored. The phage-bearing insert can then be selected to bind to the desired antigen. This method can be repeated by reselecting a round of phage that binds to the desired antigen. Repeated rounds result in enrichment of phage carrying a particular sequence. DNA sequence analysis can be performed to confirm the sequence of the expressed peptide. The smallest straight line portion of the sequence to which the antigen is desired to be bound can be determined. The process can be repeated using a library containing the insert, which contains some or all of the smallest straight line portion, plus one or more additional degenerate residues upstream, or downstream thereof. These techniques confirm the peptides of the present invention and still have a greater binding affinity for Ang-2 than the formulations already identified herein. Regardless of the manner in which the peptide is prepared, standard recombinant DNA procedures can be used to produce nucleic acid molecules encoding each of such peptides and peptibodies. The nucleotide sequence of such a DNA molecule can be appropriately manipulated without changing the amino acid sequence encoded thereby, in order to explain the degeneracy of the nucleic acid code and to explain the cryptic advantage in a particular host cell. The recombinant DNA technique is a convenient method for preparing the full length peptibody of the present invention and other large protein specific binding agents, or fragments thereof. A DNA molecule encoding a peptibody or fragment can be inserted into an expression vector, which can be inserted into a host cell in sequence to produce an antibody or fragment. In general, DNA molecules encoding peptides or peptibodies can be obtained using the procedures described herein in the Examples. Probes and typical hybridization conditions are those such as those set forth in Ausubel et al. (Current Protocols in Molecular Biology, Current Protocols Press [1994]). After the hybridization, -88- 1317281 (84)

BRIEF DESCRIPTION OF THE INVENTION Continuation _ «Si'iS can flush the detected ink dots with appropriate stringency, depending on the size of the probe, the expected homology of the probe to the pure germline, the type of library to be screened, And factors such as the number of pure lines to be screened. An example of a high stringency screen is 0.1 X SSC and 0.1% SDS at temperatures between 50-65 °C. The peptide of the present invention which binds to Ang-2 can also be confirmed by using a yeast two-hybrid screening method. Thus, antigens or fragments thereof can be used to screen peptide libraries, including phage display libraries, to identify and select Ang-2 binding agents, such as the skin of the present invention. Alternatively, various expression vector/host systems can be utilized to house and characterize the peptides of the invention. These systems include, but are not limited to, microorganisms, such as bacteria transformed with recombinant phage, plastid or plastid DNA expression vectors; yeasts transformed with yeast expression vectors; and infections transmitted by viral expression vectors (eg, baculovirus) Insect cell system; transfer infection by viral expression vector (eg, cauliflower mosaic virus, CaMV; Tobacco mosaic virus, TMV), and transfer of infected plant cell systems with bacterial expression vectors (eg, Ti or pBR322 plastids); or mammals cell. Mammalian cells useful in recombinant protein production, including but not limited to VERO cells, HeLa cells 'Chinese hamster ovary (CHO) cell line, COS cells (like COS-7), W138, BHK, HepG2, 3T3, RIN 'MDCK, A549' PC12, K562 and 293 fine monthly packages. A representative draft of recombinant expression peptides is described later. ••Performance carrier" means the plastid, bacteriophage, virus or vector to express a polypeptide from a DNA (RNA) sequence. A performance vector can include a transcription unit comprising (1) a genetic element or element that has a regulatory role in gene expression, such as a promoter or promoter sequence, and (2) a structure or sequence encoding a binding agent -89- 1317281 γ~ Τ—(85) The Ftilili column, which is transcribed into mRN A ' and then translated into protein, and (3) a collection of appropriate transcriptional start and stop sequences. The structural unit to be used in a yeast or eukaryotic expression system preferably includes a leader sequence capable of secreting the translated protein to the outside of the cell by the host cell. Alternatively, in the absence of a leader or delivery sequence, where the recombinant protein is expressed, an amine-based terminal thiomethanthine residue may be included. This residue may or may not be cleaved from the expressed recombinant protein to provide the final peptide product. For example, the peptide can be expressed in a recombinant manner in yeast using a commercially available expression system such as Pichia Expression-System (Invitrogen, San Diego, CA) according to the manufacturer's instructions. The system also relies on the pre-pro-a system to direct secretion, but the greening effect of the insert is driven by the alcohol oxidase (AOX1) promoter when induced by methanol. . The secreted peptide is purified from the yeast growth medium by, for example, a method for purifying the peptide from bacterial and mammalian cell supernatants. Alternatively, the cDNA encoding the peptide can also be cloned into the baculovirus expression vector pVL1393 (PharMingen, San Diego, CA). The vector can be used according to the manufacturer's instructions (PharMingen) to infect the Spodoptera frugiperda cells in a medium free of sF9 protein and to produce recombinant proteins. The recombinant protein can be purified and concentrated from the culture medium using a heparin-sepharose column (Pharmacia). Alternatively, the peptide can be expressed in an insect system. Insect systems for protein expression are well known to those skilled in the art. In one such system, 'Autographa California nuclear polyhedrosis - 90- 17 1317281(10) virus (AcNPV) can be used as a carrier to express foreign larvae in autumn larvae or Trichoplusia larvae. gene. The peptide coding sequence can be cloned into a non-essential region of the virus, such as the polyhedrin gene, and placed under the control of a polyhedrin initiation gene. The smooth insertion of the peptide will render the polyhedrin gene inactive and produce a recombinant virus lacking the coat protein shell. The recombinant virus can be used to infect autumn beetle cells or larvae of the larvae, and the peptide is expressed therein. Smith et al, J. Virol. 46: 584 (1983); Engelhard et al, Proc. Nat. Acad. Sci. (USA) 91:3224-7 (1994).

In other examples, the DNA sequence encoding the peptide can be amplified by PCR and cloned into a suitable vector, such as pGEX-3X (Pharmacia)» designed pGEX vector, resulting in glutathione-S encoded by the vector. a transferase (GST), and a fusion protein of a protein encoded by a DNA fragment inserted into the selection site of the vector. Primers for PCR can be produced, including, for example, suitable incision sites. The recombinant fusion protein can be cleaved from the GST portion of the fusion protein by using the fusion moiety alone to promote performance, or otherwise not wishing to attach the peptide of interest. The PGEX-3X/specific binder peptide construct was transformed into E. coli XL-1 Blue cells (Stratagene, La Jolla CA), and the individual transformants were isolated and grown. The plastid DNA from each of the transformants can be purified and partially sequenced using an automated sequencer to confirm that the nucleic acid insert encoding the desired specific binding agent is present in the appropriate orientation. Certain peptide compositions of the invention are those in which the peptibodies are conjugated to any anti-tumor peptide, such as tumor necrosis factor (TNF). In a particularly preferred method, the sequence encoding the peptide is fused to a sequence encoding TNF (Novagen, Madison, WI) in a framework to produce a TNF-specific binding agent peptide by recombinant fusion-91 - 1317281 Page] -Λ .. —· _ .. . (87) Chimera. The peptide-TNF cDNA can be cloned into the pET-llb vector (Novagen) and the TNF-peptide can be induced to express in BL21 E. coli according to the instructions of the manufacturer of pETlib. From the bacterial lysate, by ammonium sulfate preparation, hydrophobic interaction chromatography on phenyl-Sepharose 6-speed flow, ion exchange chromatography on DEAE-toolipid flow, The soluble TNF-peptide was purified by gel filtration chromatography on Sephacryl-S-300 HR. The fusion protein which can be produced in the form of insoluble inclusion bodies in bacteria can be purified as follows. Host cells can be sacrificed by centrifugation; rinsed with 0.15 M NaCl, 10 mM Tris, pH 8, 1 mM EDTA; and treated with 0.1 mg/ml lysozyme (Sigma, St. Louis, MO) for 15 minutes at room temperature. The lysate was clarified by ultrasonic oscillation and pelletized by centrifugation at 12,000 X g for 10 minutes. The fusion protein containing the pellets can be resuspended in 50 mM Tris, pH 8 and 10 mM EDTA, layered on 50% glycerol, and centrifuged at 6000 Xg for 30 minutes. The pellets can be resuspended in standard phosphate buffered saline solution (PBS) without Mg+ + and Ca+ +. The recombined protein can be further purified by fractionation of resuspended pellets (Sambrook et al., supra) in denatured SDS-PAGE. The gel can be immersed in 0.4 M KC1 to visualize the protein, which can be cut and electroeluted in a buffer solution that lacks the SDS running gel. If the GST/fusion protein produced in the bacterium is a soluble protein, GST can be cleaved from the peptide of the present invention using a GST Purification Base Unit (Pharmacia) purified digestible fusion protein. The digestion reaction can be cultured at room temperature (20-40 mg fusion protein '20-30 units human thrombin (4000 units/mg, Sigma) 'in 0.5 ml PBS) 16-48 hours-92- 1317281 88) and loaded into a denatured SDS-PAGE gel to fractionate the reaction product. The gel can be immersed in 0.4 M KC1 to visualize the protein band. The identity of the protein profile consistent with the expected molecular weight of the peptide was confirmed by an amino acid sequence analysis using an automatic sequencer (Applied Biosystems Model 473Α, Foster City, CA). Alternatively, the identity of the peptide can be confirmed by HPLC and/or mass spectrometry analysis of the peptide. Alternatively, the DNA sequence encoding the peptide can be cloned into a plastid containing the desired promoter and, if desired, a leader sequence [Better et al, Science-240: 1041-43 (1988)]. The sequence of the construct can be confirmed by automatic sequencing. The plastids can then be transformed into E. coli strain MC1061 using standard procedures using bacterial CaCl2 culture and heat shock treatment (Sambrook et al., supra). The transformed bacteria can be grown in LB medium supplemented with carboxypenicillin and can be induced to produce protein by culturing them in a suitable medium. If a leader sequence is present, the secretion of the peptide is completed and cleaved during secretion. The recombinant protein that has been secreted can be purified from the bacterial culture medium by the method described below. Mammalian host systems that represent recombinant proteins are well known to those skilled in the art. Host cell lines can be selected for processing the expressed protein, or for producing certain translational-post-modification capabilities that are useful in providing protein activity. Modifications of such proteins include, but are not limited to, acetylation, carboxylation, glycosylation, phosphorylation, lipidation, and oximation. Different host cells, such as CHO, HeLa, MDCK, 293, WI38 and their analogs, have specific -93- 1317281 for this type of translation-posterior activity.

Cellular mechanisms and unique mechanisms, and can be selected to ensure proper modification and processing of imported proteins. It is hoped that the transformed cells used are 'preferably available for long-term, high-yield protein production' and that they are inherently stable. Once the cells are transformed with vectors containing selectable markers, along with the cassettes that are desired to be expressed, they can be tolerated. The cells are grown in enhanced medium and then transferred to a selective medium to design a selectable marker that confers resistance to selection, while its presence allows for successful growth and recovery of cells expressing the introduced sequence. It is possible to use a tissue culture technique suitable for the cell to proliferate stably transformed cell clusters. A five-year selection system can be used to recover cells that have been transformed for recombinant protein production. Such selection systems include, but are not limited to, in tk-, hgprt- or aprt- cells, respectively, HSV thymidine kinase, hypoxanthine-Wulbin luciferase, and adenine to phosphoribosyltransferase gene. It is also possible to use the resistance of sputum-metabolites as the basis for the selection of DHFR, which confers resistance to amine-based sputum; gpt's conferring resistance to mycophenolic acid; neo's confers on amino-based Sugar: y: (3418's resistance and resistance to chlorsulfuron; and hygro, which confers resistance to hygromycin. Other potentially useful alternative genes, including trpB, The cells are allowed to use sputum instead of tryptophan, or hisD, which allows the cells to use histamine instead of histidine. Markers that provide visual indications for confirmation of transformants, including the flower-phone station, beta-uronic acid enzyme And its receptor GUS, and insect luciferase and its receptor luciferin. Purification and refolding of the specific sputum -94- 1317281 (90) In some cases, the specific binding agent, like The peptides and/or peptibodies of the invention may need to be "refolded" and oxidized to the appropriate tertiary structure and produce disulfide linkages to become biologically active. A number of procedures well known in the art can be used. Complete refolding. Such methods include, for example, promoting In the presence of the agent, the solubilized polypeptide formulation is exposed to a pH typically greater than 7. The choice of solubilizing agent is similar to the selection method used for inclusion body solubilization, however, the solubilizing agent is typically used at low concentrations. In most cases, the refolding/oxidizing solution will also contain a reducing agent, plus its oxidized form, in a specific ratio to produce a specific redox potential, allowing the disulfide to be shuffled. The formation of a cysteine bridge occurs. Some commonly used redox couples include cysteine/cystamine, glutathione/dithiobis GSH, copper chloride, dithiothreitol DTT/dithiane DTT, And 2-mercaptoethanol (bME) / disulfide-bME. In many cases, co-solvents can be used to increase the efficiency of refolding. Common cosolvents include glycerin, polyethylene glycols of various molecular weights, and arginine It may be desirable to purify the peptides and peptibodies of the invention. Protein purification techniques are well known to those skilled in the art. In one standard, these techniques involve the crude fractionation of protein and non-protein fractions. Peptides and/or peptibodies isolated from proteins can be further purified by chromatography and electrophoresis techniques to achieve partial or complete purification (or homogeneity purification). Particularly suitable for the preparation of the peptides of the present invention. Or the method of analysis of the peptibody is ion exchange chromatography, exclusion chromatography: polypropylene guanamine gel electrophoresis: isoelectric focusing. The present invention relates in some aspects to purification, and in particular specific - 95· 1317281 (91) In the examples, the substantial purification of the peptibody or peptide of the present invention. The term "purified peptibody or peptide" as used herein is intended to mean that it can be obtained from other components. An isolated compound in which the peptibody or peptide is purified to any grade relative to its naturally available state. Thus, a purified peptide or peptidate also means a peptibosome or peptide that is free of the environment in which it may naturally occur. In general, "purified" will mean a peptide or peptidic composition that has been subjected to fractionation, removing various other components, and which substantially retains its biological activity of expression. Where the term "substantially purified" is used, the term will mean a peptide or peptidic composition wherein the peptibody or peptide forms a major component of the composition, such as about 50% of the composition, about 60%, about 70%, about 80%, about 90%, about 95% or more protein. A variety of methods for quantifying the degree of purification of peptides or peptibodies will be apparent to those skilled in the art from this disclosure. These include, for example, determining the specific binding activity of the active fraction, or assessing the amount of the peptide or peptide in the fraction by SDS/PAGE analysis. A preferred method for assessing the purity of a peptide or peptidic fraction is to calculate the binding affinity of the dissolving portion, compare it to the binding activity of the original extract, and calculate the degree of purification in this context. Purify the number " to evaluate. The actual unit used to represent the amount of binding activity will, of course, depend on the particular assay technique selected after purification, and whether the peptide or peptide exhibits detectable binding activity. Various techniques suitable for purification will be well known to those skilled in the art. These include, for example, the precipitation by sulphuric acid, PEG, anti-body (immune sink) and the like, or by thermal denaturation, followed by centrifugation: chromatography step-96-fill face 1317281 (92), like Is affinity chromatography (eg protein-A-agarose), ion exchange, gelation, reverse phase, matrix-based limestone and affinity chromatography; isoelectric focusing; gel electrophoresis; and these and other techniques The combination. As is generally known in the art, it is believed that the order in which the various purification steps are carried out may be altered, or certain steps may be omitted, and a method suitable for preparing a substantially pure specific binder may still be produced. It is generally not necessary to always provide the peptide or peptibody of the present invention in its purest state. Indeed, it is expected that substantially less specific binder product will still be utilized in certain embodiments. Partial purification can be accomplished by using fewer purification steps in the combination, or by using different forms of the same common purification scheme. For example, 'understand that cation-exchange column chromatography using an HPLC apparatus' will generally result in more "-fold" purification than the same technique using a low pressure laminate system. A method showing a lower degree of relative purification may be advantageous for the total recovery of the peptide or peptibody or for maintaining the binding activity of the skin or skin. It is known that different conditions of SDS/PAGE can be utilized, sometimes significantly altering the movement of the peptide or celestial body [Capaldi et al, Biochem. Biophys. Res. Comm., 76: 425 (1977)]. It is therefore understood that under different electrophoresis conditions, the surface molecular weight of the product represented by the purified or partially purified specific binder can be altered. Binding Assays - Immunological binding assays typically employ a capture agent that specifically binds and often immobilizes the analyte target antigen. The capture agent is the part that is specifically combined with the analyte. In a particular embodiment of the invention, the capture agent is a peptide or peptibosome or fragment thereof that is specifically associated with Ang-2 • 97· 1317281 (93). These immunological binding assays are well known in the art [Asai, ed., Methods in Cell Biology, Vol. 37, Antibodies in Cell Biology, Academic Press, Inc., New York (1993)]. Immunological binding assays often use a labeling agent that will signal the presence of bound complexes formed by the capture reagent and the antigen. The labeling agent can be a molecule comprising a bound complex, i.e., it can be a labeled specific binding agent or an antibody labeled as an anti-specific binding agent. Alternatively, the labeling agent can be a third molecule, typically another antibody, which binds to the bound complex. The labeling agent can be, for example, an antibody carrying a labeled anti-specific binder. The second antibody, which is specific to the bound complex, may lack the label but may be bound by a fourth molecule specific to the antibody species to which the second antibody belongs. For example, a detectable moiety, such as biotin, can be used to modify the second antibody and then bind it to a fourth molecule, such as the enzyme-labeled streptavidin. Other proteins that specifically bind to the immunoglobulin constant region, such as protein A or protein G, can also be used. These binding proteins are normal components of the cell wall of streptococcal bacteria and exhibit strong non-immunogen reactivity to immunoglobulin constant regions from various species. Akerstrom, J. Immunol., 135: 2589-2542 (1985); Chaubert, Mod. Pathol., 10:585-591 (1997). In the assay, it may be necessary to have a culture and/or rinse step after each mixing of the reagents. The culturing step can vary from about 5 seconds to several hours, preferably from about 5 minutes to about 24 hours. However, the incubation time will depend on the assay format, analyte, solution volume, concentration, and the like. Usually 'measurement will be carried out at ambient temperature, although it can also be carried out at a range of temperatures -98-1317281 (94). A. Non-competitive binding assays: Immunological binding assays can be of a non-competitive type. These assays directly measure the amount of analyte captured. For example, in a preferred "three-therapy" assay, the capture reagent (antibody or peptidomimetic) can be directly bound to the solid substrate and immobilized there. These immobilized capture agents then recapture (join) the antigen present in the test sample. The protein so immobilized is then combined with a labeling agent, such as a second antibody with a label. In another preferred "sandwich" assay, the second antibody lacks a label but binds to a labeled antibody specific for the antibody species from which the second antibody is derived. A detectable moiety, such as biotin, can also be used to modify the second antibody, allowing a third labeled molecule, such as streptavidin, to specifically bind to it. See Harlow and Lane, Antibodies, A Laboratory Manual, Chapter 14, Cold Spring Harbor Laboratory, NY (1988), which is incorporated herein by reference. B. Competitive Binding Assay: Immunological binding assays can be of a competitive type. The amount of analyte present in the sample is indirectly measured by measuring the analyte added, by the analyte present in the sample, or by competing for the amount of the capture agent (antibody or peptidic body). In a preferred competitive binding assay, a known amount of analyte, usually indicated, is added to the sample and the sample is then contacted with the capture reagent. The amount of analyte bound to the antibody has been indicated to be inversely proportional to the concentration of analyte present in the sample (see Harlow and Lane, Antibodies, A Laboratory Manual, Chapter 14, pages 579-583). -99- 1317281 (95) Description of the invention: In other preferred competitive binding assays, the capture reagent is immobilized on a solid substrate. The amount of protein bound to the capture reagent can be determined by measuring the amount of protein present in the protein/antibody complex, or otherwise by measuring the amount of protein that remains uncomplexed. The protein content can be detected by providing the labeled protein. Harlow and Lane (in the first place). Another preferred competitive binding assay utilizes the inhibition of haptens. There, known analytes are immobilized on a solid substrate. A known amount of antibody is added to the sample and the sample is contacted with the analyte that has been immobilized. The amount of antibody bound to the immobilized analyte is inversely proportional to the amount of analyte present in the sample. The amount of immobilized antibody can be detected by detecting the amount of immobilized antibody or the amount of the fraction remaining in the solution. Where the antibody is labeled, it can be detected directly, or by subsequent addition of the labeled moiety, which is indirectly detected by binding to the antibody as described above. C. Utilization of Competitive Binding Assay: For the determination of cross-reactivity, a competitive binding assay can be used to allow the skilled person to determine whether the protein or enzyme complex recognized by the peptibody of the present invention is desired. The protein, rather than the cross-reactive molecule, or the peptide is determined to be antigen-specific and does not bind to an unrelated antigen. In this type of assay, the antigen can be immobilized on a solid support and an unknown protein mixture is added to the assay which will compete with the immobilized protein for binding to the peptide. Competitive molecules also bind to one or more antigens that are not associated with the antigen. The ability of a protein to compete with the immobilized antigen for binding to the peptide is compared to the binding of the same protein immobilized on a solid support to determine the cross-reactivity of the protein mixture. -100- !317281 (96) Binding assay of lion butterfly D. The present invention also provides a Western blot method to detect or quantify the presence of Ang-2 in a sample. This technique typically involves 'separating the sample protein' based on molecular weight by gel electrophoresis and moving the protein onto a suitable solid support such as nitrocellulose filter paper, nylon filter paper, or derivatized nylon filter paper. . The sample was cultured with a peptibody or a fragment thereof specifically bound to Ang-2 and the resulting complex was detected. These peptibodies can be directly labeled, or the labeled antibody can be used in combination with the peptibody for subsequent detection. Inducing a derivatized binding agent, such as the peptides and peptibodies of the invention, or fragments thereof, for diagnosing a condition or disease characterized by the expression of an Ang-2 or subunit, or for monitoring an Ang-2 to be utilized Determination of e Ang_2 by the inducer, fragment, Ang_y, agonist or inhibitor treated patient, including the use of peptibodies and markers to detect Ang- in human body fluids or cells or tissue extracts 2 method. Peptides of the invention with or without modification can be used. In a preferred diagnostic assay, the body will be labeled by attaching, for example, marking or reporting the φ molecule. Various labels and reporter molecules are known, and some of them have been described herein. The present invention is particularly useful for diagnosing human diseases. A variety of drafts for measuring Ang-2 protein using peptibodies specific for each protein are known in the art. Examples include enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), and Rauma-activated cell sorting (FACS). The two-site, single-strain-based immunoassay is preferably -101 - 1317281 (97), which utilizes a single antibody that reacts with two non-contradictory epitopes on Ang-2, but Competitive binding assays can be used. These assays are described, for example, in Maddox et al, J. Exp. Med., 158: 121 1 (1983). To provide a basis for diagnosis, normal or standard values for human Ang-2 are usually established. It is known in the art that, under conditions suitable for complex formation, a body fluid or cell extract derived from a normal individual, preferably a human, is mixed with a peptide body against Ang-2. This determination is completed. The amount of standard complex formation can be quantified by comparing the control and disease samples with the binding of the peptibody to a known amount of Ang-2 protein-mass. The standard value obtained from the normal sample can then be compared to the value obtained from a sample from an individual who may be affected by the disease. The bias between the standard and individual values suggests the role of Ang-2 in disease states. For diagnostic applications, in certain embodiments, the peptibodies or peptides of the invention will typically be labeled with a detectable moiety. The detectable portion can be any portion that can produce a detectable signal directly or indirectly. For example, the detectable moiety can be a radioisotope such as a 3H, 14C, 32P, 35S or 1251 'fluorescent or chemiluminescent compound such as luciferin isothiocyanate, rhodamine or luciferin; Or an enzyme such as alkaline phosphatase, beta-galactosidase or horseradish peroxidase. Bayer et al., Meth. Enz., 184: 138-163 (1990). Diseases The present invention provides binding agents that bind to Ang-2, such as peptides, peptibodies' or fragments, variants or derivatives thereof, which are useful for treating human diseases and pathological conditions. A preparation for modulating Ang-2 binding activity or other cellular activity may be mixed with other therapeutic agents to enhance the therapeutic effect or reduce the side effect of the -102-(98)1317281* method of the present invention Eight 1^-2 live. Situation, like increase and infertility. The present invention also encompasses the low Ang-2 activity method for the inclusion of 'included'. The method consists of phase-measuring preferably: or reducing the cancer's fine-tuned adaptation to the cancer cell assay. Can be borrowed from this material. Lactation: or companion animals and rabbits, also tumor cells and cargo tissue cells called malignant benign growth aspects provide reagents and diseases that can be used to treat diseases and conditions. The condition is characterized by unwanted or deviant disease in the cells. Including cancer, as well as other hyperproliferative conditions, psoriasis, contact dermatitis, immunological disorders, providing a method of treating cancer in animals including humans, and having a prostitute - g pt. ta. A specific amount of a specific binding agent, such as a peptide that inhibits or degrades the present invention, which is directed against a cell line that inhibits the growth of cancer cells, and the "process of cell proliferation, invasiveness, and metastasis" using the present invention as a cancer cell growth. In inhibitor activities, silly θ like 疋 mammals use this method to inhibit cell growth, invasion, metastasis or tumor incidence. It can also be used quickly in the measurement system, for example, growth and its characteristics, and confirmation of the growth of cancer cells.

Cancers treated by the method of the moon, preferably occur in mammals, including humans and other primates, as well as pets, such as hyenas and cats, laboratory animals, such as rats, rats, and farms. Animals, like horses, pigs, sheep and cattle. The growth of cells in the biological sentence # 丄 i is uncontrolled and progressing. Some of these growths are benign, but will cause other organisms to die. A malignant neoplasm or cancer differs from that in addition to showing aggressive cell proliferation, it -103 - 1317281

(99) SMiS may invade surrounding tissues and transfer. In addition, malignant neoplasms are characterized by their loss of more differentiation (more dedifferentiation) and their organization relative to each other and their surrounding tissues. This property is also referred to as "regressive development." The neoplasms that can be treated by the present invention also include solid tumors, i.e., carcinomas and sarcomas. Cancers include those malignant neoplasms derived from epithelial cells that infiltrate (invade) the surrounding Tissue, and cause metastasis. Adenocarcinoma is a cancer derived from glandular tissue, or it forms a recognizable glandular structure. Other widespread species or cancer: Symptoms, including sarcomas, which are tumors, whose cells are buried In fibrils or homogeneous substances, such as the connective tissue of an embryo. The invention is also capable of treating cancers of the bone marrow or lymphatic system, including leukemias, lymphomas, and other forms that are usually not in the form of tumor masses, but are scattered throughout the blood vessels. Or a cancer in a lymphatic network. The type of cancer or tumor cell that can be treated according to the present invention includes, for example, a tumor producing ACTH, acute lymphocytic leukemia, acute non-lymphocytic leukemia, adrenal cortical cancer, bladder cancer, brain Cancer, breast cancer, cervical cancer, chronic lymphocytic leukemia, chronic myeloid leukemia', colorectal cancer, skin T-cell lymphoma, endometrial cancer, esophageal cancer, Ewing's sarcoma, gallbladder cancer, hairy cell leukemia, head and neck cancer, Hodgkin's lymphoma, Kaposi's (KaposPs) sarcoma, kidney cancer, liver cancer, lung cancer (small and non-small cells), malignant peritoneal effusion' malignant pleural effusion, melanoma, mesothelioma, multiple myeloma, neuroblastoma, glial Tumor, non-Hodgkin's lymphoma, osteosarcoma, ovarian cancer, ovarian (germ cell) cancer, pancreatic cancer, penile cancer, pre-104-1317281 (100) adenocarcinoma, retinoblastoma, skin cancer , soft tissue sarcoma, squamous cell carcinoma, gastric cancer, testicular cancer, thyroid cancer, trophoblastic neoplasm, uterine cancer, vaginal cancer, vulvar cancer and nephroblastoma.

The invention is particularly explained herein with respect to certain types of treatments for experimentally defined cancers. In these interpretive treatments, standard in vitro and in vivo modes have been used with the highest standards available. These methods can be used to confirm that it is expected to be an effective preparation in a therapeutic regimen in vivo. However, it will be appreciated that the methods of the invention are not limited to the treatment of these tumor types, but extend to any solid tumor from any organ system. Cancers that are invasive or metastasically associated with Ang-2 expression or activity are particularly susceptible to inhibition by the present invention or even cause regression. The present invention can also be practiced by including a compound of the present invention, such as a peptibosome, in combination with other anti-cancer chemotherapeutic agents, such as any conventional chemotherapeutic agent. The combination of a proprietary binder with such other formulations can produce the effectiveness of a chemical treatment draft. Many chemotherapeutic drafts will present themselves in the light of the artist's ideas and can be incorporated into the methods of the present invention. Any chemotherapeutic agent can be used, including alkylated formulations, anti-metabolites, hormones and antagonists, radioisotopes, and natural products. For example, the compound of the present invention may be mixed with an antibiotic such as doxorubicin and other anthracycline analogs, such as a ring-shaped amine, a striking analog like a 5-fluorouracil bite. , cisplatin, basal vein, taxol, and natural and synthetic derivatives thereof, and analogs thereof. Like other examples, in the case of mixed tumors, such as breast adenocarcinoma, where the tumor includes gonadotropin-dependent and gonadotropin-independent cells, it can be connected -105 - 1317281

(ιοί) The compound is administered together with leuprolide or gosereiin (a synthesis of LH-RH). Other anti-tuberculosis drafts include the use of tetracycline compounds and other therapeutic regimens, such as surgery, radiation, etc., also referred to herein as "assisted anti-caries medications." Thus, the method of the present invention can be used with this The use of a traditional-like regimen together has the advantage of reducing side effects and increasing efficacy. Thus the invention provides compositions and methods that can be used to treat various cancers, including solid tumors and leukemias. Types of treatable cancer, including but not limited to : adenocarcinoma of the breast, prostate and colon; all forms of lung · bronchial cancer; bone marrow; melanoma; liver tumor; neuroblastoma: papilloma; amine precursor uptake and decarboxyloma; Neoplasms; primary tumors, malignant carcinoid syndrome; carcinoid heart disease; cancer (eg Walker, basal cells, basal squamous cells, Brown-Pearce, catheter's 'Ire District' Ehrlich) tumor, Krebs 2, Merkel cells, mucin, non-small cell lung, oat cells, papillary Hard cancer, bronchiole, bronchial, squamous and transitional cells; tissue cell disorders; leukemia; malignant tissue cell proliferation 'Hodgkin' disease; immune sputum small lung squamous cell carcinoma; Hodgkin's lymphoma; plasmacytoma; reticuloendotheliosis; melanoma; chondroblastoma, chondroma; chondrosarcoma; fibroids; fibrosarcoma; giant cell tumor; histiocytoma; Liposar sarcoma; mesothelioma; myxoma; mucinous sarcoma, osteoma; osteosarcoma; chondroma; craniopharynx; dysgerminoma; hamartoma; mesenchymal; middle renal tumor; Neoplasm: gingival blastoma; dental tumor; teratoma; thymoma; gout -106- 1317281

(102)

A tophoblastic tumor. In addition, it can also treat the following types of cancer: adenoma; cholangiocarcinoma; cholesteatoma; cylindrical tumor; cystadenocarcinoma; cystadenoma; granulosa cell tumor; amphoblastic blastoma; hepatic neoplasm: sweat gland adenoma ;Island cell tumor; Leydig cell tumor; papilloma; Sertoli cell tumor; follicular cell tumor; leiomyomas; leiomyosarcoma; myoblastoma; fibroids; Rhabdomyosarcoma; rhabdomyosarcoma: ependymoma; ganglionoma; glioma; neural tube blastoma; meningioma; schwannomas; neuroblastoma; neuroepithelial neoplasia; neurofibromatosis; neuroma; Ganglio; non-chromophobic paraganglioma; vascular keratomas; vascular lymphoid hyperplasia with eosinophilia; sclerosing hemangioma; blood tuberculosis; glomus tumor; hemangioendothelioma; Vascular sarcoma; angiosarcoma; lymphangioma; lymphangioma; lymphangiosarcoma; pineal tumor; carcinosarcoma; chondrosarcoma; phyllodes sarcoma; fibrosarcoma; angiosarcoma; leiomyosarcoma; leukemia sarcoma Liposarcoma; Lin bar tube sarcoma; myosarcoma; myxosarcoma; ovarian cancer; rhabdomyosarcoma; sarcoma; neoplasms; disease, neurofibromatosis, and cervical dysplasia.

Another aspect of the present invention is the use of the materials and methods of the present invention to prevent and/or treat any hyperproliferative condition of the skin, including cowhide dancing and contact dermatitis, or other hyperproliferative diseases. Patients with kraft and contact dermatitis have been shown to have elevated Ang 2 activity in these lesions [Ogoshi et al., j. inv. Dermatol" 1 10:818-23 (1998)]. Ang-2-specific single-agent combination with other pharmacological agents is used to treat people who exhibit these clinical symptoms. Any of a variety of planting agents can be used, as described herein, and others who are familiar with this Well-known -107- 1317281

(103) The route of administration to deliver a specific binding agent. Other aspects of the invention include the treatment of various retinopathy involving angiogenesis (including urinary retinopathy and age-related macular degeneration), and conditions/diseases of the female genital tract, such as endometriosis , uterine fibroids, and other conditions associated with vascular proliferative dysfunction (including endometrial microvascular growth) during the female reproductive cycle. Other aspects of the present invention relate to the treatment of abnormal blood vessel growth, including cerebral arteriovenous malformations (AVMs), gastrointestinal mucosal injury and repair, in patients with a history of peptic ulcer disease, ulceration of the gastroduodenal mucosa, including Ischemia in the hair, in patients with non-hepatic portal hypertension, extensive pulmonary vascular disease in patients with liver disease and portal hypertension. Another aspect of the invention is the use of the compositions and methods provided by the present invention to prevent cancer. Such agents will include a specific binding agent, such as a peptide against Ang-2. Pharmaceutical composition

Ang-2 specific binding agents, such as pharmaceutical compositions of peptibodies, are within the scope of the invention. The pharmaceutical compositions include the antibodies described in detail in U.S. Patent No. 6,171,586, issued to Jan. Such compositions include a therapeutically or prophylactically effective amount of a specific binding agent, such as an antibody or fragment, variant, derivative or fusion protein thereof as described herein, and in pharmaceutically acceptable form The preparation is mixed. In a preferred embodiment, the pharmaceutical composition comprises an antagonistic, specific binding agent that partially or fully modulates the biological activity of at least one Ang-2 and is combined with a pharmaceutically acceptable formulation. Typically, the specific binding agent will be purified for administration to the animal's to be filled with -108- 1317281 _ (104) I €13⁄43⁄4⁄4 parts. The pharmaceutical composition may contain a formulation to modify, maintain or retain, for example, the pH, permeability, viscosity, clarity, color, isotonicity, taste, sterility, stability, dissolution or release, absorption or absorption of the composition. Rate of penetration "Appropriate formulations include, but are not limited to, amino acids (such as glycine, glutamine, aspartame, arginine or lysine); antimicrobial agents; antioxidants ( Such as ascorbic acid, sodium sulfite or sodium bisulfite); buffer solutions (such as borate, bicarbonate, Tris-HCl, citric acid, salt, phosphate; other organic acids); fillers (like mannitol) Or glycine); a chelating agent [like ethylenediaminetetraacetic acid (EDTA)]; a complexing agent (such as caffeine, polyvinylpyrrolidone, β-cyclodextrin or hydroxypropyl-β-cyclodextrin); Fillers; monosaccharides; disaccharides and other carbohydrates (like glucose, mannose or dextrin); proteins (like serum albumin, gelatin or immunoglobulin); pigments; flavors and diluents; Hydrophilic polymer (such as polyvinylpyrrolidone); low molecular weight peptide; salt-forming counterion (like sodium); preservatives (such as benzalkonium chloride, alum acid, salicylic acid, ethylmercury water) Sodium salicylate, phenylethyl alcohol, methyl acetonate, propyl chlorate, chlorhexidine, sulphuric acid or hydrogen peroxide; solvent (like glycerol, propylene glycol or polyethylene glycol) ); sugar alcohols (like mannitol or sorbitol); suspending agents: surfactants or wetting agents (like pluronics, PEG, sorbitan esters, polyethoxylated chains) , such as polysorbate 20, polysorbate 80, tee, trimethylamine tromethamine, lecithin, cholesterol, tyloxapal; stability enhancer (sucrose or Sorbitol); a tonicity enhancer (such as alkali gold-109-1317281 (105) is a functional (preferably sodium or potassium chloride), mannitol, sorbitol); a delivery vehicle; a diluent; Excipients and/or pharmaceutical adjuvants qRemington, s Pharmaceutical Sciences, 18th edition, edited by AR Gennaro Mack Publishing Company, 1990). The most suitable pharmaceutical composition will be exemplified by, for example, the intended route of administration, the delivery format, and the desired dosage. See, for example

Remington’s Pharmaceutical Sciences ’ previous. Such compositions can affect the physical state, stability, release rate of the left in vivo - and the rate of clearance in vivo - in pharmaceutical compositions, the main vehicle or compliment, J flies The nature of the agent may be aqueous or non-aqueous. For example, a suitable vehicle $% of the pate carrier can be water for injection, saline solution or artificial cerebrospinal lumbar nights. It may be supplemented with other often used in parenteral compositions. && history Substance neutral buffered saline or saline mixed with serum albumin is a representative vehicle. Other representative pharmaceutical compositions include Tris buffer solution of approximately 8.5 Å, or pH 4.0. -5.5 acetate buffer solution, which is a substitute. In the present invention comprising sorbitol or a composition thereof having a desired, desirable purity and 16 cases, the 'sperm will be selected. .,, the formulation of the agent (Remington’s

Pharmaceutical Sciences, formerly)), Kunjin. , ,. ^ ^ .^ 〇 'Prepare a cold-drying cake or a water-filled night, to prepare a misplaced gentleman

0 to oxime compound © In addition, a suitable excipient such as sucrose may be used, # D W ^ 4, , , '. The mixture product is formulated into a lyophilizate. A pharmaceutical sputum can be selected for parenteral administration for inhalation or enteral delivery, as appropriate. Or 'optional combination 吟疋口月 g per ear, meridian, transrectal -no. !317281

(106) or transvaginal. The preparation of such pharmaceutically acceptable compositions is within the skill of the art. The formulation component is present at a concentration that is acceptable for administration of the site, for example, using a buffer solution to maintain the composition at a physiological pH, or a slightly lower pH value - typically at a pH ranging from about 5 to about 8. Inside. In the case of an attempted parenteral administration, the therapeutic composition for use in the present invention may be in the form of a pyrogen-free, parenterally acceptable aqueous solution, in a pharmaceutically acceptable vehicle, including the desired -Binding agent. Particularly suitable: The enteroinjected vehicle is a sterile distilled water in which the binding agent is formulated into a sterile, isotonic solution suitable for storage. Another article may involve a nucleating molecule such as a center ft, bioerodable particles, polymerized rainbow, polyglycolic acid, bead or vesicles such as a squid, which provides The control or sustained release of the product is ~ storage injection delivery. It is also possible to use the effect of the transparency of the 11th and the second period. Other guides u have implanted drug delivery devices in the circulation. The method 'package in other aspects' can be sputum in aqueous solution, and is suitable for parenteral injection. Λ Μ 乐 调 调 , , , , , , , , , , , , , , , 生理 生理 生理 生理 生理 生理 生理 生理(Hank's) solution, Ringer's solution or physiologically like a solution. Aqueous injection suspensions may contain an aqueous suspension; the buffer may be buffered like sodium carboxymethylcellulose, sorbitol or dextran. Or: a suspension of the substance's compound is prepared into a suitable oily injection, which can be a live lipophilic solvent or vehicle, including a fat oil, like a different taste of the oil, a gas-and-sour vinegar, like The oleic acid, the triglyceride or the lipid can also be delivered using the -111 - 1317281 (107) amine-based polymer of non-lipid polycations. The suspension may also contain suitable stabilizers as needed, or a formulation which increases the solubility of the compound, and allows for the preparation of very high concentrations of solutions. In other embodiments, a pharmaceutical composition for inhalation can be formulated. For example, the binding agent can be formulated into a dry powder for inhalation. A propellant can also be used to formulate an inhalation solution of a polypeptide or nucleic acid molecule for aerosol delivery. In another embodiment, the solution can be atomized. Lung administration is further described in PCT Application No. PCT/US94/001875, which describes the delivery of a chemically modified protein to the lungs. It has also been attempted to orally administer certain formulations. In a particular embodiment of the invention, the binding agent molecules administered in this manner may or may not be formulated with those carriers conventionally used in the mixing of solid dosage forms, such as lozenges and capsules. For example, the capsule can be designed to release the active portion of the formulation somewhere in the gastrointestinal tract, with the highest bioavailability and the lowest pre-systemic degradation. Additional formulations may be included to promote absorption of the binding agent molecules. Diluents, flavoring agents, low melting point mash, vegetable oils, lubricants, suspending agents, tablet disintegrating agents and binders can also be used. Pharmaceutical compositions for oral administration can also be formulated using pharmaceutically acceptable carriers well known in the art, at dosages suitable for oral administration. Such carriers are capable of formulating pharmaceutical compositions into tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like for ingestion by a patient. A pharmaceutical composition for oral use can be obtained by mixing the active compound with a solid excipient and processing the resulting mixture of granules (after grinding as needed) to obtain a lozenge or dragee core-112-1317281 (108) heart. . If necessary, add the appropriate adjuvant. Suitable excipients include hydrophobic compounds or protein fillers such as sugars including lactose, sucrose, mannitol and sorbitol; starches derived from corn, wheat, rice, potato or other plants; cellulose, such as Cellulose, hydroxypropylmethyl-cellulose or sodium carboxymethylcellulose; gums, including gum arabic and tragacanth; and proteins such as gelatin and collagen. If desired, a disintegrating or solubilizing agent such as crosslinked polyethylene-pyrrolidone, agar and alginic acid, or a salt thereof may be added, such as sodium sulphate. A sugar-coated core may be used in conjunction with a suitable coating film, such as a concentrated sugar solution, which may also contain gum arabic, talc, polypyridyl pyrrolidone, carbopol gel, polyethylene glycol and/or titanium dioxide, A lacquer solution and a suitable organic solvent or solvent mixture. Dyestuffs or pigments may be added to the key or coating film to confirm the product or to indicate the content characteristics of the active compound, i.e., the dosage. Pharmaceutical products which can be used orally also include capsules made of push gel, as well as soft, sealed capsules made of gelatin, and coatings such as glycerol or sorbitol. The push-fit capsules may contain the active ingredient in admixture with a filler or binder such as lactose or starch, a lubricant such as talc or magnesium stearate, and stabilizers if desired. In soft capsules, the active ingredient may be dissolved or suspended in a suitable liquid, such as a fatty oil, liquid, or liquid polyethylene glycol, with or without a stabilizer. Other pharmaceutical compositions may involve an effective amount of a binding agent in a mixture with a non-toxic excipient suitable for the manufacture of a tablet. A solution in unit dosage form can be prepared by dissolving the tablet -113- 1317281 (109) [SMll in sterile water, or other suitable vehicle. Suitable excipients include, but are not limited to, inert diluents - such as calcium carbonate, sodium or sodium bicarbonate, lactose or calcium phosphate; or binders such as starch, gelatin or gum arabic; or lubricants, Like magnesium stearate, stearic acid or talc. Additional pharmaceutical compositions will be apparent to those skilled in the art' including formulations involving the formulation of binding agent molecules in a sustained or controlled-delivery manner. Techniques for various other sustained- or controlled-release methods, such as _ is a liposome carrier, bioerodible microparticles, or porous beads and reservoir injections are also known to those skilled in the art. See, for example, pct/us93/00829' for the controlled release of porous polymeric microparticles for delivery of pharmaceutical compounds. Other examples of sustained-release articles, including semi-permeable polymeric substrates', are in the form of shaped articles, such as films or microcapsules. The matrices released by the performance may include polyacetate 'water colloid, polylactide (U.S. Patent No. 3,773,919 'European Patent No. 58,481), copolymer of L-proline and γ-ethyl-glutamate [Sidman et al. Human, Biopolymers, 22: 547-556 (1983)], poly(2-hydroxyethyl-isobutyl citrate) [Langer et al., j. Biomed. Mater. Res., 15: 167-277, (1981) )] and [Langer et al., Chem. Tech., 12: 98-105 (1982)] 'Ethylene vinyl acetate (Langer et al., former) or poly-D(-)-3-hydroxybutyric acid (Europe) Patent No. 133, 988). The sustained-release composition also includes aliquots which can be prepared by any of a number of methods known in the art. See, for example, Eppstein et al., Proc. Natl. Acad. Sci. (USA), 82: 3688-3692 (1985); European Patent No. 36,676: European Patent No. 88,046; European Patent No. 143,949. -114- 1317281 (110) Pharmaceutical compositions intended for in vivo administration must generally be sterile. This can be achieved by filtration through a sterile filtration membrane. Where the composition is lyophilized, the sterilization using this method can be carried out before or after the drying and reconstitution of the cold east. The parenteral compositions can be stored in lyophilized form or in solution. In addition, the parenteral compositions are typically placed in a container with a sterile joint, such as an intravenous solution bag, or a vial having a stopper pierceable by a hypodermic needle. Once the pharmaceutical composition has been formulated, it can be stored in a sterile vial in the form of a solution, suspension, gel: gel, emulsion, solid, or dehydrated or lyophilized powder. Such formulations may be stored in ready-to-use form or in the form of reconstitution (e.g., freeze-drying) prior to administration. In a particular embodiment, the invention is directed to a kit that produces a single-dose administration unit. The kit may contain a first container having dehydrated protein and a second container having an aqueous formulation, respectively. Also included within the scope of the invention is a kit of prefilled syringes (e.g., liquid syringes and lyosyring) containing single and multiple-separate compartments. The effective amount of the pharmaceutical composition to be used in therapy will depend, for example, on the deficiencies and goals of the treatment. Those skilled in the art will know the dosage level appropriate for the treatment, and therefore will depend in part on the molecule to be delivered, the indications of the molecule to be used, the route of administration, and the size (weight, body surface area or organ size) and condition of the patient (age) And change in general health). Because of this, the clinician can titrate the dose and modify the route of administration to achieve the best therapeutic effect. Representative dosages can range from about 0.1 mg/kg to up to about 100 mg/kg or more, depending on the factors mentioned above -115 - 1317281 _ on) \fmmn. In other embodiments, the dosage may range from 0.1 mg/kg up to about 100 mg/kg; or 1 mg/kg up to about 100 mg/kg; or 5 mg/kg up to about 100 mg/kg. For any compound, a therapeutically effective dose can be established initially in a cell culture assay, or in an animal model such as a mouse, rat, rabbit, dog, pig or monkey. Animal models can also be used to determine the appropriate concentration range and route of administration. This information can then be used to determine the dose and route of administration that is useful in humans. The exact dose will be determined from the factors associated with the individual in need of treatment. The dosage and administration are adjusted to provide a sufficient level of active compound or to maintain the desired effect. Factors that may be considered include the severity of the disease state, the general health of the individual, the age, weight and sex of the individual, the timing and frequency of administration, the combination of drugs, the sensitivity of the response, and the response to treatment. Long-acting pharmaceutical compositions can be administered every 3 to 4 days, weekly or biweekly, depending on the half-life and clearance rate of the particular formulation. The frequency of administration will depend on the pharmacokinetic parameters of the binding agent molecules used in the formulation. The composition is usually administered until the dose at which the desired effect is achieved is reached. Thus, the composition can be administered in a single dose or in multiple doses (at the same or different concentrations/doses)' or in a continuous infusion over a period of time. The appropriate dosage will be discussed routinely in more detail. Appropriate doses can be probed by using appropriate dose-response data. The pharmaceutical composition is administered according to known methods, such as oral administration, intravenous injection, intraperitoneal, intracerebral (intrinsic), intraventricular, intramuscular, intraocular, intraarterial, intraportal. , the path within the lesion -116- 1317281 (112) diameter, spinal cord, intrathecal, intraventricular, percutaneous, subcutaneous, intraperitoneal, intranasal, enteral, local, sublingual, urethra, vagina or rectum, By continuous release of the system, or by implanting the device. Where desired, the composition can be administered by bolus injection or continuous infusion, or by implantation. Additionally or additionally, the composition may also be topically administered via implantation of a film, sponge, or other suitable substance in which the desired molecule has been adsorbed or encapsulated. Where an implanted device is used, the device can be implanted into any suitable tissue or organ and the desired molecule can be delivered via diffusion, on-time release of the mass, or continuous administration. In some cases, it may be desirable to use a pharmaceutical composition in a manner that is in vitro. In such instances, the cells, tissues or organs that have been removed from the patient are exposed to the pharmaceutical composition, and the cells, tissues and/or organs are subsequently transplanted back into the patient. In other cases, the binding agents of the present invention can be delivered by implanting certain cells that have been genetically engineered to express and secrete the polypeptide, such as those described herein. Peptide. Such cells may be cells of an animal or human and may be autologous, xenogeneic or xenogeneic. The cells can be immortalized as needed. To reduce the change in the immunological response, the cells can be encapsulated to avoid infiltration of surrounding tissue. The encapsulated material is typically a biocompatible, semi-permeable polymeric encapsulant or membrane that permits release of the protein product (s), but prevents the cell from being affected by the patient's immune system, or by other tissues from the surrounding tissue. Harmful damage. Combination Therapy The specific binding agents of the present invention, such as peptibodies, can be used in conjunction with other therapeutic agents that treat diseases associated with the performance of Ang-2-117-1317281 (113). These other therapeutic agents include, for example, Radiation therapy, chemotherapy, and targeting therapy, such as HerceptinTM, Ritux.a_nTM, GleevecTM, and the like. Other combinations of treatments not specifically recited herein are also within the scope of the invention. Chemotherapy may use anti- and sputum biological preparations, including, for example, alkylating preparations, including nitrogen mustard, such as mechlorethamine, cycloamine, isosfamide, amphetamine ( Melphalan) and chlorambucil; sub-nitrourea, such as nitroso mustard (BCNU), lomustine (CCNU) and semustine (semustine) Methyl-CCNU); aziridine/methyl melamine, such as tri-ethyl melamine (TEM), tri-ethyl thiophosphonamide (thiotepa), hexamethyl honey Amines (HMM, hexamethylene melamine); alkyl sulfonates such as busulfan; triad 11 such as dacarbazine (DTIC); antimetabolites including folic acid analogues, Such as amine mites and trimetrexate, "····························································· Pyrimidine glycoside] 5-Azacytidine, 2,2'-difluorodeoxycytidine, anthraquinone analogs, such as 6-mercaptopurine, 6-thiopurine, nitro-purine, 2'-deoxy-assisin (pentostatin), erythrohydroxynonyladenine (EHNA), Fludarabine phosphate, and 2-chlorodeoxy Adenine (cladribine, 2-CdA); natural products, including anti-mitotic drugs, such as paclitaxel, periwinkle bioassay, including periwinkle test (VLB), Changchun new test and vinorelbine宾 (vinorelbine), smear cancer, estramustine and estramustine citrate; epipodophyllotoxins, like 依118-1317281

(114) fmMM topo '^ (etoposide) and teniposide, antibiotics like actinomycin D, daunorubicin (erythromycin), doxorubicin, mitoxantrone (mitoxantrone) ), idarubicin, broccoli, plicamycin (photo-mycin) lysin C and actinomycin, enzymes like L-aspartate; Biological response modifiers like 'interferon-a, IL_2, G-CSF and GM-CSF; miscellaneous formulations' include platinum coordination complexes such as cisplatin and carboplatin, anthracenedione Like the Mito E from Kun's substituted urea, like hydroxyurea, methyl hydrazine derivatives, including N-methyl hydrazine: (MIH) and procarbazine 'adrenal cortex inhibitors, like Mitotane (o,p'-DDD) and aminoglutethimide; hormones and antagonists, including adrenal corticosteroid antagonists like dehydrocortisone and equivalents, dexamethasone and Aminolutamine; a corpus luteum preparation, such as hydroxyprogesterone caproate, medroxyprogesterone acetate, and Megestrol acetate; estrogen, such as diethylstilbestrol and ethinyl estradiol; antiestrogens, like tamoxifen; androgen, These include sterolone propionate and fluoxymesterone/equivalent; antiandrogens such as flutamide, gonadotropin-releasing hormone analogues and leuprolide; and anti-steroid resistance Androgen, like flutamide. A combination of growth factors, including cytokinins, lymphokines, growth factors, or other hematopoietic factors such as M-CSF, GM-CSF, TNF, IL-1, IL-2, IL-3, IL- 4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-14, IL-15, IL-16, IL-17-119- 1317281 (115), IL-18, IFN, TNFO, TNF1, TNF2, G-CSF, Meg-CSF, GM-CSF, thrombopoietin, stem cell factor and erythropoietin. Other compositions may include known 'angiogenins, such as Ang-1, -2, -4, -Y, and/or human Ang-like polypeptides, and/or vascular endothelial growth factor (VEGF). Growth factors include angiogenin, bone morphogenetic protein-1, bone morphogenetic protein-2, bone morphogenetic protein-3, bone morphogenetic protein-4, bone morphogenetic protein-5, bone morphogenetic protein -6, bone morphogenetic protein-7, bone morphogenetic protein-8, bone morphogenetic protein-9, bone morphogenetic protein-10, bone morphogenetic protein-11, bone morphogenetic protein-12, bone morphogenetic protein -13, bone morphogenetic protein-14, bone morphogenetic protein-15, bone morphogenetic protein receptor IA, bone morphogenetic protein receptor IB, brain-derived neurotrophic factor, cilia neutrophil, cilia Neutral factor receptor, cytokinin-induced neutrophil chemokine 1, cytokinin-induced neutrophil chemokine 2, cytokinin-induced neutrophil chemokine 2 , endothelial cell growth factor, endothelin 1, epithelial growth factor, epithelial-derived neutrophil attractor, fibroblast growth factor 4, fibroblast growth factor 5, fibroblast Long factor 6, fibroblast growth factor 7, fibroblast growth factor 8, fibroblast growth factor 8b, fibroblast growth factor 8c, fibroblast growth factor 9, fibroblast growth factor 10, fibroblast growth Factor acidic, fibroblast growth factor alkaline, glial cell line-derived neutrophil receptor-1, glial cell line-derived neutrophil receptor-2, growth-related protein Growth-related protein-1, growth-related 13172281 (116) protein-2, growth-related protein-3, heparin-binding epidermal growth factor, hepatocyte growth factor, hepatocyte growth factor receptor, insulin-like Growth factor I, gonadotropin-like growth factor receptor, insulin-like growth factor Π, insulin-like growth factor binding protein, keratinocyte growth factor, leukemia inhibitory factor, leukemia inhibitory factor receptor-1, nerve growth factor, nerve growth Factor receptor, neurotrophin-3, neurotrophin-4, placental growth factor, placental growth factor 2, platelet Endothelial growth factor, platelet-derived growth factor, platelet-derived 'long-factor A chain, platelet-derived growth factor AA, platelet-derived growth factor AB, platelet-derived growth factor B chain, platelet-derived growth Factor BB, platelet-derived growth factor receptor-1, platelet-derived growth factor receptor-2, pre-B cell growth stimulating factor, stem cell factor, stem cell factor receptor, transforming growth factor-1, transforming growth factor-2 , transforming growth factor-1, transforming growth factor-1.2, transforming growth factor-2, transforming growth factor-3, transforming growth factor-5, latent transforming growth factor-1, transforming growth factor-1 binding protein I, transforming growth factor -1 binding protein II, transforming growth factor-1 binding protein III, tumor necrosis factor receptor type I, tumor necrosis factor receptor type I, urokinase-type plasminogen activator receptor, vascular endothelium Growth factors, as well as chimeric proteins, and fragments thereof that are biologically or immunologically active. Immunotherapy " Immunotherapy usually relies on the use of immune effector cells and molecules that target and destroy cancer cells. The immune effector can be, for example, a peptibody of the invention that recognizes some of the markers on the surface of the target cell. Peptibodies can be used alone as -121 - 1317281 (117) er er er \ \ Λ Λ Λ * * * Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Λ Peptidom bodies can also be combined with drugs or toxins (chemotherapeutic agents, radionuclides, ricin Α chain, Vibrio cholerae toxin, pertussis toxin, etc.) and are therefore primarily targeted. According to the present invention, a mutant form of Ang-2 can be obtained by the peptibody or peptidomimetic of the present invention by immunotherapy. It is particularly desirable to use the peptidomimetic compositions of the present invention in a combination therapy with Ang-2 targeting therapy.

Passive immunotherapy has proven to be particularly effective for many cancers. See, for example, W0 98/39027. The following examples are for illustrative purposes only and are not to be construed as limiting the scope of the invention in any way. Example 1 Ang-2 expression in pathological sputum and normal tissues The performance of Ang-2 was examined in normal and pathological tissues using in situ hybridization. Humans (Genbank accession number: AF004327, nucleotides 1274-1726) and mice (Genbank accession number: AF004326' nuclear station acid 1 135-1588) were amplified from human or mouse fetal lung cDNA by reverse transcriptase-PCR. Fragments of the φ Ang-2 sequence were cloned into the pGEM-T plastid and confirmed by sequencing. From the linear plastid template, the 33P-labeled antisense RNA probe was transcribed using 33P-UTP and RNA polymerase. Formaldehyde-fixed, paraffin-embedded tissue blocks were cut into 5 microns and collected on charged slides. Prior to in situ hybridization, 0.2 M HCl was first passed through the tissue, then digested with proteinase K and acetylated with triethanolamine and acetic anhydride. The sections were hybridized overnight with a radiolabeled probe at 55 ° C, then subjected to RNase digestion, and • 122- rnmm 1317281 (118) at 55 ° C in a high stringency wash in approximately 〇丨X SSC. Slides were immersed in Kodak NTB2 emulsion and exposed for 2-3 weeks at 4eC' development and contrast staining. Examination of sections with dark field and standard illumination 'allows simultaneous assessment of histomorphology and hybridization signals. The results indicate that in normal postnatal humans, the performance of Ang-2 is limited to a small number of tissues containing vascular blood vessels, such as the ovary, placenta, and uterus. There is no detectable Ang-2 expression in the heart, brain, kidney, liver, lung, pancreas, spleen 'muscle, tonsils, thymus', appendix, lymph nodes, gallbladder, anterior gland or sputum in normal adults. In 5-week old rats (but not in adult monkeys or humans), the kidneys exhibited significant Ang-2 performance in straight tubes. To determine whether the performance is a regenerative embryo, the expression was repeated using the old Ang-2 probe and the above conditions on the kidneys derived from mice up to 1 year old. It was observed that the performance of Ang-2 was reduced during the development of neonates but was still evident in the kidneys of the 1 year old machine. In almost all of the tested tumor types, the performance of Ang-2 was detected, including the original. Human tumors like 'colon cancer (5 cases), breast cancer (10 cases), lung cancer (8 cases), glioblastoma (1 case), metastatic human tumors, like breast cancer (2 Cases), lung cancer (2 cases) and ovarian cancer (2 cases), which have metastasized to the brain's and caries-like tumor models, such as C6 (rat glioma), HT29 (human colon cancer), Colo-205 (human colon cancer), HCT116 (human colon cancer), A43 1 (human epidermoid carcinoma), A673 (human rhabdomyosarcoma), HT1080 (human fibrosarcoma), PC-3 (human prostate cancer), B16F10 ( Mouse melanoma), MethA (rat sarcoma) -123- 奋 说 办 ] ] ] ] 1317281 (119) and Lewis (Lewis) lung cancer. Furthermore, VEGF was also expressed in new blood vessels in the Matrigel plug, and Ang-2 expression was detected in a hypoxic model of precocious retinopathy in mice. Example 2 Assessing the molecular weight of Ang_2 month

Molecular assays (affinity ELISA, neutralization ELISA and BIAcore) were developed to evaluate the peptibodies that bind directly to Ang-2 and related family members, and the effect of peptibodies on the interaction of Ang-2: Tie-2. Describe these assays in vitro - as follows.

Affinity ELISA For the initial screening of candidate anti-Ang-2 peptibodies, purified human Ang-2 (R&D Systems, Inc.; catalogue 623-AN; supplied as a mixture of 2 truncated versions) ANg-2), or mouse Ang-2 polypeptide (prepared as described above). For confirmatory binding assays, human Ang-2 obtained from human 293T cell conditioned medium was transfected with full length human Ang-2 DNA and contained no serum, containing approximately 50 μg/ml bovine serum albumin (BSA). Dulbecco's was modified in modified Eagle medium (DMEM). Using a microtiter plate, add approximately 100 microliters of Ang-2 to each well and incubate the plate for approximately 2 hours, then rinse the plate 4 with phosphate buffered saline (PBS) containing approximately 0.1% Tween-20. Times. The wells were then blocked using approximately 250 liters/well of approximately 5% BSA in PBS and the plate was incubated for approximately 2 hours at room temperature. After incubation, the excess blocking solution was discarded and started at a concentration of approximately 40 nanomoles, followed by PBS containing approximately -124- 1317281 (120) -.7 -1 r ' ΐ.1 1% BSA Serial dilutions were serially diluted 4-fold and approximately 100 microliters of each candidate anti-Ang-2 peptibody was added to the wells. The plate was then incubated overnight at room temperature. After incubation, the plate was rinsed with PBS containing approximately 1% Tween-20. Repeat the rinsing four times, then add 100 μl/well to goat anti-human IgG (Fc)-HRP (Pierce Chemical Co., catalog #31416) pre-treated according to 1:5000 in PBS containing 1% BSA. . The plate was incubated at room temperature for about 1 hour. It was then washed 5 times with PBS containing approximately 0.1% Tween-20 followed by approximately 100 microliters/well of TMB (3,33,5^tetramethylbenzidine liquid receptor system; Sigma Chemical Company, St. Louis, MO, catalogue No. T8665) was conditioned and cultured for approximately 5-15 minutes until the blue color was revealed. The absorbance is then read at a spectrophotometer of approximately 370 nm.

Neutralization ELISA A microtiter plate that has bound human Ang-2 polypeptide was prepared as described by affinity ELISA. Candidate anti-Ang-2 peptibodies, from 1000 nM to 0.2 pM, containing approximately 1% BSA and approximately 1 nM Tie-2 (provided as a Tie-2-Fc molecule, wherein the Tie-2 portion only contains The soluble extracellular portion of the molecule; R&D Systems, catalogue No. 3 13-TI) was diluted 4-fold in PBS. After adding 100 μl of antibody/Tie-2 solution to each well, the plate was incubated overnight at room temperature and then washed 5 times with PBS containing about 0.1% Tween-20. After rinsing, approximately 100 microliters/well of anti-Tie-2 antibody (Pharmingen Inc., catalog #557039) was added to a final concentration of approximately 1 μg/ml and the plate was incubated for approximately 1 hour at room temperature. Next, approximately 100 μl/well of goat anti-mouse-IgG-HRP (Pierce Chemical Co., catalog #31432) was added using a 1:10,000 dilution with PBS containing approximately 1% BSA. Incubation at room temperature -125- 1317281 hair 谏 谏 竟 j j:-two' id-' (121) The plate was incubated for about 1 hour and then washed 5 times with PBS containing approximately 0.1% Tween-20. Approximately 100 μl/well of TMB substrate (described above) was then added and allowed to appear in color. The absorbance was then read at 370 nm in a spectrophotometer. Affinity BIAcore

Affinity analysis of each candidate Ang-2 peptibody was performed on BIAcore® 2000 (Biacore, Inc., Piscataway, NJ) using PBS and 0.005% P20 surfactant (Biacore, Inc.) as the execution buffer solution. The recombinant protein G (Repligen, Needham, MA) was immobilized on a research grade CM5 sensor wafer (Biacore, via a primary amine group using an amine coupling kit (Biacore, Inc.) according to the manufacturer's proposed draft. Inc.).

By first capturing approximately 100 Ru of each candidate anti-Ang-2 peptibody to the already immobilized protein G, various concentrations (0-100 nM) of huAng-2 were then applied at a flow rate of 50 μl/min. Binding assays were performed by injection of mAng-2 onto the surface of the bound antibody for 3 minutes. The BIA estimates 3.1 computer program (Biacore, Inc.) was used to determine the kinetic parameters of peptidomimetic binding, including ka (association rate constant), kd (dissociation rate constant), and KD (dissociation equilibrium constant). The lower the dissociation equilibrium constant, the greater the affinity of the peptibody for Ang-2. Example 3

Confirmation of Ang-2 Binding Peptide 1. Preparation of Magnetic Beads Coated with Ang-2 A. Fix Ang-2 on Magnetic Beads for non-specific elution, select rounds for all three, for every 100 micro The mother liquor of the beads obtained from the manufacturer contains about 4 μg of biotinylated Ang-2 protein concentration, and the biotinylated Ang-2 protein (Biotin-126-1317281 (122) ; a recombinant human angiopoietin-2, R&D Systems, Inc.; catalogue BT623) immobilized in Streptomyces avidin Dynabeads (Dynal, Lake)

Success, NY). For the elution of antigen (Ang-2) and receptor (Tie-2), 2 μg of biotinylated Ang-2 protein was immobilized on 50 μl of Streptomyces avidin Dynabeads for two selection rounds . For the third selection round, reduce the coating concentration to 50 μl of bead mother liquor, containing approximately! Microgram of biotinylated Ang-2 protein. By using a magnet to attract the beads to the side of the tube and sucking off the liquid, the phosphate buffered physiological salt -

Water (PBS) was rinsed 5 times and resuspended in PBS. The concentration of the above-described Ig-2 protein was added to the washed beads and incubated at room temperature for 1 hour, and then rotated, followed by incubation at 4 ° C for several hours, and Rotate and then block the Ang-2 coated beads by adding BSA to a final concentration of approximately 1%, incubate at 4 ° C, spin and then rinse with PBS before accepting the selection procedure The resulting Ang 2 coated beads were 5 times. B. Preparation of beads for negative selection Additional beads were also prepared for negative selection. For each panning condition, 500 microliters of the mother bead from the manufacturer was subjected to the above procedure (paragraph 1A) except that the step of culturing the biotinylation, 2 was omitted. In the final rinse step, the beads are divided into 5 aliquots of microliters. 2. Select Ang-2 binding 0^ bacteria release._ A. Comprehensive strategy using three filamentous sputum libraries, named "ΤΝ8_Ιχ" (5χι〇9 independent transformants), "TNU-r (my independent transformants) and "straight line" -127- 1317281 (123) (2.3X109 independent transformants) (all from Dyax c〇rp.) to select Ang-2 binding bacillus . Each pool was then subjected to non-specific elution, Ang-2 washout and receptor elution (Tie-2). For Ang-2, 9 different panning conditions were used (TN8-IX using non-specific elution method, TN8-IX using Ang_2 elution method, TN8-IX using Tie-2 elution method, use) Non-specific wash. Detachment of TN12-1, TNI2-1 using Ang-2 elution method, TN12-1 using Tie-2 elution method 'Line using non-specific elution method, using Ang -2 straight line of the elution method, and a straight line using the Tie-2 elution method). For all three pools, the phage selected from the first round of the round was eluted only in a non-specific manner for the next round of selection. Elution was performed using Ang-2 and Tie-2 in the selection of the second and second rounds. As for the linear library, only the second round of Ang-2 and Tie-2 elution options were selected. B. Negative selection For each panning condition, approximately 100 random pool equivalents of the TN8-IX and TN12-I pools were used from the stock solution (TN8-IX was approximately 5 X 1011 pfu, while TN12-I was approximately 1.4 X 1〇11 pfu and approximately 1 random pool equivalent of the linear library (approximately 1 X 10npfu) were divided into aliquots and diluted to approximately PBST (PBS with 〇.〇5% 吐温吁-20) 400 μl. After the last rinse, beads prepared from the initial 100 μl aliquot for negative selection (the liquid was aspirated in the paragraph, and approximately 400 μl of the diluted stock solution was added to the beads. At room temperature The resulting mixture was incubated for about 10 minutes and spun. The phage supernatant was aspirated using a magnet and added to the second 100 microliter aliquot for additional negative selection steps. Thus, 5 negatives were performed. Selection procedure -128- 1317281 (124) C. Selection of beads coated with Ang-2 protein After the last negative selection step (paragraph 1B), the phage supernatant was added to the Ang-2 coated small Beads (paragraph 1A). Incubate the mixture for 1 to 2 hours at room temperature and With rotation, the phage was allowed to bind to the target protein. After discarding the supernatant, the beads were rinsed with PBST approximately 10 times, followed by rinsing twice with PBS. D. Non-specific elution in aspirating the final rinse liquid (paragraph 2C), after adding a large-about 1 ml of Min A salt solution (60 mM K2HP〇4, 33 mM ΚΗ2Ρ04, 7.6 mM (NH4) S04 and 1.7 mM sodium citrate) to the beads. Add to concentrated bacterial samples for infection (see paragraphs 3A and 3B). E. Adhered to the phage antigen (Ang-2) for the second round, after the final rinse step (paragraph 2C), By continuously adding 100 μl of 1 pM, 0.1 nM and 10 nM recombinant Ang-2 protein (recombinant human angiopoietin-2, R&D Systems, Inc. Minneapolis, Minnesota), each condition was incubated for 30 minutes. The bound phage is eluted from the magnetic beads. The remaining phage are eluted in a non-specific manner (paragraph 2D). The phage eluted from the 10 nM and non-specific elution are mixed, and Make it accept the choice of round 3 (see paragraph 4 below). After 3 rounds, after the final rinse step (paragraph 2C), approximately 1 nM of recombinant Ang-2 protein and 10 nM of recombinant Ang-2 protein were continuously added, each condition was incubated for 30 minutes from magnetic beads. The bound phage was eluted. Further, the bacillus was eluted with 1 ml of a 1 mM solution of triethylamine (Sigma, St. Louis, Missouri) for 10 minutes on a rotator. -129- 1317281 |_ (125) 丨_Description_ The pH of the phage-containing triethylamine solution was neutralized with 0.5 ml of 1 M Tris-HCl (pH 7.5). After the last elution with 100 mM triethylamine solution, the remaining phage was eluted by adding beads to the bacteria (paragraph 2D). F. The reporter of the combined phage (Tie-2) elutes

Regarding the second round, after the final rinsing step (paragraph 2C), approximately 100 μl of 1 pM, 0.1 nM and 10 nM recombinant Tie-2 protein (recombinant human Tie-2-Fc in combination) was continuously added. R&D Systems, Inc. Minneapolis, Minnesota), each condition was incubated for 30 minutes, eluting from magnetic beads: bound phage. The remaining phage were eluted in a non-specific manner (segment 2D). Phage eluted from 10 nM and non-specific elution were mixed and allowed to accept the selection of round 3 (see paragraph 4 below). For the third round, after the final rinse step (paragraph 2C), approximately 1 nM of recombinant Ang-2 protein and 10 nM of recombinant Tie-2 protein were continuously added, each condition was incubated for 30 minutes, from magnetic The bound phage is eluted in the beads. Further, on the rotator, the bacillus was eluted with 1 ml of a 100 mM triethylamine solution (Sigma, St. Louis, Missouri) for 10 minutes. The pH of the phage-containing triethylamine solution was neutralized using 0.5 ml of 1 M Tris-HCl (pH 7.5). After the last elution with 100 mM triethylamine solution, the remaining phage was eluted by adding beads to the bacteria (paragraph 2D). 3. Expanding effect A. Preparation of tiled cells Fresh E. coli (XL-1 Blue MRF') cultures were grown to an OD600 of approximately 0.5 in LB medium containing approximately 12.5 μg/ml tetracycline. For each panning condition, approximately 20 ml of the culture was cooled on ice -130-1317281 (126). The bacterial pellet was resuspended in approximately 1 ml of a solution of Min A salt. B. Transduction A mixture of each of the different elution methods described above (paragraphs 2D, 2E and 2F) was added to a concentrated bacterial sample (Section 3A) and cultured at approximately 37 ° C for approximately 15 minute. About 2 ml of NZCYM medium (2X NZCYM, 50 μg/ml ampicillin) was added to each mixture and incubated at about 37 ° C for 15 minutes. The resulting 4 ml solution was plated on a large NZCYM agar plate containing: about 50 μg/ml ampicillin and incubated overnight at 37 °C. C. Harvesting of phage Each bacterial/phage mixture was grown overnight on large NZCYM agar plates (Section 3B) and subsequently scraped into approximately 35 mL of LB medium. The agar plates were further washed with an additional 35 ml of LB medium. The resulting bacterial/phage mixture in LB medium was centrifuged to remove the pellet formed by the bacteria. Approximately 50 ml of the phage supernatant was then transferred to a new tube and approximately 12.5 ml of PEG solution (20% PEG 8000, 3.5 M ammonium acetate) was added and incubated on ice for 2 hours to pellet the phage. The precipitated phage was centrifuged and resuspended in 6 ml of phage resuspension buffer (250 mM NaCl, 100 mM Tris pH 8, 1 mM EDTA). The remaining bacteria were removed by centrifugation, the phage solution was further purified, and the phage was precipitated twice by adding about 0.5 ml of a PEG solution. After the centrifugation step, the phage pellets were resuspended in approximately 400 microliters of PBS. The solution was subjected to final centrifugation to remove any residual bacterial debris solution. Standards for Use -131 - 1317281 Description of the Invention Continued: ί (127) Determination of plaque formation, titration of the resulting phage preparation. 4. Additional selection and expansion In the second round, the expanded phage preparation from the first round (approximately 101 (3 pfu) was used as the input phage for the selection and expansion steps (paragraphs 2 and 3). As for Ang- 2 and Tie-2 solution, mixed with 10 nM and non-specifically eluted phage, and expanded for the selection of the third round. Then use the expanded phage product from the second round (about 109 pfu) as Input

The cells were selected and expanded in the third round (paragraphs 2 and 3). After the elution step of the third round: (paragraphs 2D, 2E and 2F), the small lysate of the eluted phage was tiled and assayed for plaque formation (paragraph 3C). Individual plaques were selected and placed in 96-well microtiter plates containing 100 microliters of TE buffer solution per well. These primary plates were incubated overnight at 4 °C, allowing the phage to elute into the TE buffer solution. 5. Selection analysis Phage pure lines were analyzed by phage ELISA and DNA sequencing. The sequences are arranged based on the combined results obtained from these two assays. A. 嗟 bacteria ELISA Φ

'Let the culture of XL-1 Blue MRF' grow until OD_ reaches approximately 0.5. The culture was divided into aliquots of approximately 30 microliters in each well of a 96-well microtiter plate. Approximately 10 microliters of eluted phage (paragraph 4) was added to each well and allowed to infect the bacteria for approximately 15 minutes at room temperature. Approximately 100 microliters of LB medium containing approximately 12.5 micrograms per milliliter of tetracycline and approximately 50 micrograms per milliliter of ampicillin was added to each well. The microtiter plate was then shaken overnight at approximately 37 °C. Allowing recombination at approximately 4 °C • 132·1317281 (128) Inventive, cont.; Ang-2 protein (approximately 1 μg/ml in PBS) was combined with a 96-well Maxisorp culture plate (NUNC) overnight. Streptomyc avidin was coated on another Maxisorp disk at approximately 2 μg/ml in PBS as a control group. On the second day, the liquid in the protein coated Maxisorp tray was discarded and utilized at approximately 4 ° C for approximately 300 μl of 5 ° /. The milk solution blocked each well overnight (or 1 hour at room temperature). The milk solution was then discarded and the wells were rinsed 3 times with PBST solution. After the final rinse step, approximately 50 microliters of pBST-4% milk was added to each well of the white-coated Maxisorp disk. Approximately 50 microliters of the overnight culture from each well in a 96-well microtiter plate was transferred to an Ang-2 coated plate and the corresponding well of the control plate of the abalone-resistant avidin plate 100 μL of the mixture in each of the plates was incubated at room temperature for about 1 hour. Discard the liquid from the #自]ylaxisorp disk and rinse each well approximately 3 times with PBST. The jjRp-my anti-antibody (Amersham Pharmacia Biotech) was diluted to approximately 1:7,500, and approximately 100 microliters of the diluted solution was added to each well of the Maxisorp disk. 'Cultivate at room temperature for about 1 hour. 'And rinse each well approximately 5 times with PBST. Approximately microliters of sigma was then added to each well and the reaction was carried out using approximately 50 microliters of 5N H2S04 solution. The OD450 was read on a spectrophotometer (Molecular Devices). Q «The sequencing of pure strains of bacterial cells 1 For each phage pure line, a sequencing template was prepared using PCR. Fragments of approximately 500 nucleotides of nuclear acid were amplified using pairs of oligonucleotides in the τ column. 5'-CGGCGCAACTATCGGTATCAAGCTG-3, (SEQ ID NO: 1372881 (129): 54) Introduction 2: S'-CATGTACCGTAACACTGAGTTTCGTC-S1 (Sequence 1J Identification: 5 5) Prepare the following mixture for each pure line: Reagent volume (microliter) / test tube dH20 26.25 50% glycerol 10 10 X PCR buffer solution (no MgCl2) 5 25 mM MgCl2 4 10 mM dNTP mixture 1 100 μΜ primer 1 0.25 100 μΜ primer 2 0.25 Taq polymerase 0.25 phage in TE (Paragraph 4) 3 Final reaction volume 50

For PCR, the following procedure was performed using a thermal cycler (GeneAmp PCR System 9700, Applied Biosystems): 94 ° C for 5 minutes: (94 ° C for 30 seconds, 55 ° C for 30 seconds, 72 ° C for 45 seconds) X 30 rounds 72 ° C for 7 minutes; cooled to 4 ° C. The QIAquick Multiwell PCR Purification Kit (Qiagen) was used to purify the PCR product from each reaction according to the manufacturer's draft. The purified PCR product was then analyzed by running a 1% agarose gel with 1 microliter of dye (10X BBXS agarose loaded dye) for approximately 1 microliter of each of the PCR reaction mixtures. The remaining products were then sequenced using the ABI 377 sequencer (Perkin Elmer) according to the manufacturer's proposed draft. • 134· 1317281 (130) Description of the invention ^ page 6. Determination of sequence alignment and consensus sequence A. Sequence alignment and analysis The peptide sequence translated from the variable nucleotide sequence (paragraph 5B) is coherent with the ELISA data. A pure line showing a high OD45 enthalpy in the pores coated with Ang-2 and a low OD450 value in the pores coated with streptavidin was preferred. It is also given priority to the sequence in which multiple occurrences occur. Based on these criteria, candidate sequences are selected for further analysis of peptides or peptibodies.

B. Determination of Sequences Three different types of consistent subjects were produced from the TN8-IX library as follows: KRPCEEXWGGCXYX (Sequence Recognition: No. 56)

KRPCEEXFGGCXYX (sequence recognition: No. 57) XXXCXDXYWYCXXX (sequence identification: No. 61) XXXCXDXYTYCXXX (sequence identification: 62) XXXCXDXFWYCXXX (sequence identification: 63) XXXCXDXFTYCXXX (sequence identification: 64) XXXCXWDPWTCEXM (sequence identification: 58) 'From The TN12-1 library produces a consistent theme: WSXCAWFXGXXXXXCRRX (sequence recognition: 59) For all consistent subject sequences, by determining the most frequently occurring amino acids in each position, the addition of each consistent sequence is obtained. The "core amino acid sequence" of the subscript line. "X" means any naturally occurring amino acid. In the TN8-IX and TN12-I libraries, the two cysteine acids adjacent to the core sequence are immobilized amino acids. -135 - 1317281 (131 BRIEF DESCRIPTION OF THE INVENTION Continued page: Peptides that bind to Ang-2 are identified in Table 3 below.

Table 3: Λπ5-2Ι吉合月A

Peptide SEQ ID NO: TN8-8 1 KRPCEEMWGGCNYD TN8-14 2 HQICKWDPWTCKHW TN8-Conl 3 KRPCEEIFGGCTYQ TN8-Con4 4 QEECEWDPWTCEHM TN12-9 5 FDYCEGVEDPFTFGCDNH LI 6 KFNPLDELEETLYEQFTFQQ C17 7 QYGCDGFLYGCMIN Example 4

Construction of DNA encoding peptibodies It is possible to select modified peptides that bind Ang-2:Tie-2 (see Table 3) to construct fusion proteins in which the monomer of each peptide or the column of each peptide is The polymer (with a linker between the monomer units) is fused in the framework to the DNA encoding the linker followed by the Fc region of human IgG1. By binding a pair of oligonucleotides (oligos), generating a polynucleotide that encodes the peptide, along with the linker, constructs each modified peptide, which will include five glycine residues depending on the peptide. , 8 glycine residues or 1 lysine residue: These constructs are produced as Ndel to Xhol fragments. These double-stranded polynucleotides were ligated into a vector containing the human Fc gene (pAMG21-Fc N-terminal, as further described below), which had previously been digested with Ndel and Xhol. The resulting ligation mixture was transformed into E. coli strain 2596 cells (GM221, further hereinafter referred to as 1317281 (132), entitled: Continuation; page: j description) by electroporation using standard procedures. Pure lines are screened for the ability to produce recombinant protein products with gene fusion of the correct nucleotide sequence. For each modified peptide, a single such pure line (i.e., Fc-peptide fusion product) was selected. Construction t>AMG21-Fc N-terminal carrier DAMG21

The expression plastid 卩8?4021 (into 1' (: (: 98113) is derived from the expression vector pCFM1656 (ATCC No. 69576), and the expression carrier system is described in US Patent No. 4,710,473, which has been published The procedure described in International Patent Application WO 00/24782 (see the section of Example 2, from pages 100 to 103, and Figures 17A and 17B).

Fc N-terminal carrier

An Fc N-terminal vector was created using E. coli strain 3788, pAMG21 Tpo_Gly5_Fc monomer as a template. Information on the selection of this line can be found in WO 00/24782 (see Example 2 and Figure 10 herein). The 5' PCR primer was designed (described further below), the Top peptide sequence in pAMG Tpo Gly5 was removed and replaced with a polylinker containing the ApaLI and Xhol positions. Using strain 3788 as a template, PCR was performed using Expand Long Polymerase, using the following sequence identification: oligodeoxynucleotide No. 8 as a 5' primer, and a versatile 3' primer, sequence identification below: No. 9 . The resulting PCR product was gel purified and digested with restriction enzymes Ndel and BsrGI. The spin column was gel purified using Qiagen (Chatsworth, CA) and the polynucleic acid and plastids encoding the peptide of interest and its linker were purified by gel. The plasmid and insert were then ligated using standard ligation procedures and the resulting ligation mixture was transformed into E. coli cells (line 2596). Select a single pure line and perform DNA sequencing. Confirmation of the correct pure line, and -137- 1317281 Description of the Sulfide] (133) Used as a carrier source for the modified peptides described herein. 54 scorpion: ACAAACAAACATATGGGTGCACAGAAAGCGGCCGCAAAAA AACTCGAGGGTGGAGGCGGTGGGGACA (sequence i line another ij: 8) 33 scorpion: GGTCATTACTGGACCGGATC (sequence identification: 9)

In addition to making these modified peptides N-terminal, fused to Fc (N-terminal peptibodies), some of them may also be C-terminal fusion products (C-terminal-peptibodies). A vector for performing C-terminal fusion is described below. Construction of Fc C-terminal carrier

The Fc C-terminal vector of the modified peptide was produced using E. coli strain 3728, pAMG21 Fc_Gly5_Tpo monomer as a template. Information on the selection of this line can be found in W0 00/24782 (see Example 2 and Figure 7 in this article). The 3' PCR primer (SEQ ID NO: 10) was designed, the Τρο peptide sequence was removed, and it was replaced with a multi-linker containing the ApaLI and Xhol positions. Using the strain 3728 as a template, PCR was carried out using an extended length polymerase using a versatile 5' primer (sequence recognition: No. 11), and the 3' primer mentioned above. The resulting PCR product was purified by gelatinization and digested with restriction enzymes BsrGI and BamHI. The spin column was purified via Qiagen gel to gel-purify both the plastid and the multi-nuclear acid encoding each of the peptides of interest and their linkers. The plastid and insert were then ligated using the standard ligation protocol and the resulting ligation mix was transformed into cells of E. coli (line 2596). A single pure line is selected and subjected to DNA sequencing. The correct pure line is confirmed and used as a carrier source for the modified peptides described herein. -138- Invented the key thin system! 1317281 (134) 53 scorpion: CGTACAGGTTTACGCAAGAAAATGG (sequence ΐ ΐ: 10) 331 sub: TTTGTTGGATCCATTACTCGAGTTTTTTTGCGGCCGCTTTCTG TGCACCACCACCTCCACCTTTAC (sequence identification: 11) GM221 (#2596^). Host line used to express Fc-peptide fusion proteins

#2596, is an E. coli K-12 strain modified to contain the lux promoter gene and containing the temperature sensitive lambda repressor cI857s7 in the early ebg region and the lacIQB repressor in the late ebg region. The emergence of these two repressor genes allows the host to use a wide variety of expression systems. The ATCC named the line 202174. Example 5 Production of Peptidom

It is expressed in the bacteria of the large intestine. Cultures of each of the pAMG21-Fc fusion constructs in E. coli were grown in Terrific Broth medium (see Tartof and Hobbs, at 37 ° C) to grow plastid and adherent pure lines. Improved media for growing plasmid and cosmid clones", Bethesda Research Labs Focus, Vol. 9, page 12, 1987, referred to in Sambrook et al., supra, incorporated above. Inducing agent, N-(3-oxohexyl)-DL-homserine lactone, to a final concentration of 20 ng/ml per ml to complete the induction of gene product expression from the luxPR promoter gene The culture was incubated for an additional 6 hours at 37 t. The bacterial culture was then examined by microscope for the presence of inclusion bodies and collected by centrifugation. In the medium of induction - 139 - 1317281 〇/. t:.. -:-.i (135) Inclusion of the refracted inclusion body, indicating that Fc-fusion is most likely to be produced in insoluble fragments in E. coli by resuspension in 10% β - Laemmli sample of mercaptoethanol In the buffer solution, the cell pellet was directly lysed and then analyzed by SDS-PAGE. In most cases, a band of the appropriate molecular weight of Coomassie-stained was observed on an SDS-PAGE gel.

purification. The cells were disrupted by high pressure homogenization (passed at 14,000 psig twice) in water (1/10) and harvested by centrifugation (4000 RPM in a J-6B centrifuge for 1 hour). . The inclusion bodies were dissolved in 6 Μ胍, 50 mM Tris, 10 mM DTT, pH 8.5 for 1 hour at a ratio of 1/10. For the linear peptide fused to Fc, the solubilized mixture was diluted 25-fold in 2 guanidine, 50 mM Tris, 160 mM arginine, 2 mM cysteine, pH 8.5. Oxidation was allowed to proceed at 4 ° C for 2 days, allowing the formation of disulfide-linked compounds (i.e., Fc-peptide homodimers). As for the cyclic peptide fused to Fc, according to the same draft, the following three folding conditions were added: (1) 2 guanidine, 50 mM Tris, 160 mM arginine, 4 mM cysteine, 1 mM cyst amine,

pH 8.5; (2) 4 guanidine, 20% glycerol, 50 mM Tris, 160 mM arginine, 2 mM cysteine, pH 8.5; and (3) 4 guanidine, 20% glycerol, 50 mM Tris, 160 mM arginine, 4 mM cysteine, 1 mM cystamine, pH 8.5. The refolded protein was dialyzed against 1.5 Μ veins, 50 mM NaCl, 50 mM Tris, pH 9·0. The pH of the mixture was lowered to pH 5 using acetic acid. The precipitate was removed by centrifugation and the supernatant was adjusted to pH 5 to 6.5 based on the isoelectric point of each fusion product. The protein was filtered and loaded into a SP-Sepharose HP column equilibrated at 20 mM NaAc, 50 mM NaCl at a pH determined for each construct at 4 °C. The 20-column volume was used, ranging from 50 mM NaCl -140 - invented to the performance page; 1317281 (136) to a linear gradient of the same buffer solution of 500 mM NaCl's eluted protein peaks were collected and filtered. Peptides produced using the above procedure are set forth in Table 4 below. Peptide Peptide Sequence L1(N) MGAQKFNPLDELEETLYEQFTFQQLEGGGGG-Fc (SEQ ID NO: 12) LI (N) WT MKFNPLDELEETLYEQFTFQQLEGGGGG-Fc (SEQ ID NO: 13) LI (N) 1KWT MKFNPLDELEETLYEQFTFOQGSGSATGGSGSTASSGS GSATHLEGGGGG-Fc (Sequence Recognition: No. 14) 2xLl (N) MGAQKFNPLDELEETLYEQFTFQQGGGGGGGGKFNPL DELEETLYEQFTFQQLEGGGGG-Fc (SEQ ID NO: 15) 2xLl (N) WT MKFNPLDELEETLYEQFTFQQGGGGGGGKFNPLDELEE TLYEQFTFQOLEGGGGG-Fc (SEQ ID NO: 16) Con4 (N) MGAQQEECEWDPWTCEHMLEGGGGG-Fc (Sequence recognition: 17) Con4 (N) 1K -WT MQEECEWDPWTCEHMGSGSATGGSGSTASSGSGSATH LEGGGGG-Fc (sequence recognition: 18) 2xCon4 (N) IK MGAQQEECEWDPWTCEHMGSGSATGGSGSTASSGSGS ATHQEECEWDPWTCEHMLEGGGGG-Fc (sequence recognition: 19) L1(C) M-Fc-GGGGGAOKFNPLDELEETLYEQFTFQQLE (sequence identification: 20)

-141 - Description of the invention: Ears: 1317281 (137) LI(C)1K M-Fc- GGGGGAQGSGSATGGSGSTASSGSGSATHKFNPLDELE ETLYEQFTFQQLE (Sequence recognition: 21) 2xLl (C) M-Fc- GGGGGAQKFNPLDELEETLYEOFTFOOGGGGGGGGKF NPLDELEETLYEQFTFQQLE (Sequence recognition: 22) Con4 (C M-Fc-GGGGGAQQEECEWDPWTCEHMLE (SEQ ID NO: 23) Con4(C) IK M-Fc- GGGGGAQGSGSATGGSGSTASSGSGSATHQEECEWDP WTCEHMLE (Sequence recognition: No. 24) 2xCon4 (C) IK M-Fc- GGGGGAQQEECEWDPWTCEHMGSGSATGGSGSTASS GSGSATHQEECEWDPWTCEHMLE (Sequence recognition: 25) Con4 -Ll (N) MGAQEECEWDPWTCEHMGGGGGGGGKFNPLDELEET LYEOFTFOQGSGSATGGSGSTASSGSGSATHLEGGGGG-Fc (Sequence recognition · · 26) Con4-Ll (C) M-Fc- GGGGGAQGSGSATGGSGSTASSGSGSATHKFNPLDELE ETLYEQFTFQQGGGGGQEECEWDPWTCEHMLE (Sequence recognition: 27) TN-12-9 (N) MGAQ-FDYCEGVEDPFTFGCDNHLE-GGGGG- Fc (SEQ ID NO: 28) C17(N) MGAQ-QYGCDGFLYGCMINLE-GGGGG-Fc (SEQ ID NO: 29) TN8-8 (N) MGAQ-KRPCEEMWGGCNYDLEGGGGG-Fc (SEQ ID NO: 30) TN8-14 (N) MGAQ-HQICKWDPWTCKHWLEGGGGG-Fc (sequence recognition: 31) Coni (N) MGAQ-KRPCEEIFGGCTYQLEGGGGG-Fc (SEQ ID NO: 32) In Table 4, "Fc" means the human Fc IgG1 sequence. Column 2 states the amino acid sequence of the peptibodies. Its Fc portion is labeled with "Fc" and is as identified in the sequence below: Statement No. 60. You should know where to use the markup, such as "Con4" or "Con-4", which means the Con-4 peptide, but use the suffix '"C", "((3)" or "-(^"; or ',_^", ''(1^)" or "-1^"' indicates that the molecule is as in the -142- invention description page 1 1317281 (138) The peptibodies described herein. The suffix "quoting N", "(N) " or" in the peptidic name indicates that the Ang-2-binding peptide (or peptide) is an N-terminal to Fc functional site, The suffix "C", "(C)" or "-C" indicates that the Ang-2-binding peptide (or peptide) is a C-terminal to Fc functional site. In addition, 2xCon4 (C) 1K As defined in Sequence Identification: No. 25, it can also be expressed in the absence of "1K". The amino acid sequence of the Fc portion of each peptidom is as follows (from the amine end to the carboxyl end):

DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWDV

SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL

HQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPS

RDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVL

DSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLS LSPGK (Sequence recognition: No. 60) The DNA sequences encoding peptibodies corresponding to peptidic sequence recognition: 12-32 (in Table 4) are hereinafter described (sequence recognition: No. 33-53): 庠 column identification :3 3

ATGGGTGCACAGAAATTCAACCCGCTGGACGAACTGGAAGAAACTCT

9$

GTACGAACAGTTCACTTTCCAGCAGCTCGAGGGTGGAGGCGGTGGGG

ACAAAACTCACACATGTCCACCTTGCCCAGCACCTGAACTCCTGGGGG

GACCGTCAGTTTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGAT

CTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGA

AGACGCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGC

ATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTAC

CGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGC

AAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATC

GAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGT

GTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAG

CCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGA

GTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTC

CCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGT

GGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGAT

GCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTC

TCCGGGTAAATAATGGATCC -143· 1317281 Announcement of Invention (1) Column Identification: No. 3 4

ATGAAATTCAACCCGCTGGACGAACTGGAAGAAACTCTGTACGAACA

GTTCACTTTCCAGCAGCTCGAGGGTGGAGGCGGTGGGGACAAAACTCA

CACATGTCCACCTTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGT

TTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACC

CCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGA

GGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAA

GACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA

GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACA

AGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCA

TCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTG

CCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGC

CTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGC

AATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGA

CTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAG

CAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGC

TCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAA

ATAA queue identification: 3 5

ATGAAATTCAACCCGCTGGACGAACTGGAAGAAACTCTGTACGAACA

GTTCACTTTCCAGCAGGGATCCGGTTCTGCTACTGGTGGTTCCGGCTCC

ACCGCAAGCTCTGGTTCAGGCAGTGCGACTCATCTCGAGGGTGGAGGC

GGTGGGGACAAAACTCACACATGTCCACCTTGCCCAGCACCTGAACTC

CTGGGGGGACCGTCAGTTTTCCTCTTCCCCCCAAAACCCAAGGACACC

CTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTG

AGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGT

GGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACA

GCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGC

TGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCA

GCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGA

ACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAA

CCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACAT

CGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGA

CCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAA

GCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCAT

GCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCC

TCTCCCTGTCTCCGGGTAAATAA -144- Description of invention Continued 丨 1317281 (140) 庠 column identification: 3 6

ATGGGTGCACAGAAATTCAACCCGCTGGACGAACTGGAAGAAACTCT

GTACGAACAGTTCACTTTCCAGCAGGGTGGTGGTGGTGGTGGCGGTGG

TAAGTTCAACCCACTGGATGAGCTGGAAGAGACTCTGTATGAACAGTT

CACTTTCCAGCAACTCGAGGGTGGAGGCGGTGGGGACAAAACTCACA

CATGTCCACCTTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTTT

TCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCC

CTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAG

GTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAG

ACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAG

CGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAA

GTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCAT

CTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGC

CCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCC

TGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCA

ATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGAC

TCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGC

AGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCT

CTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA

TAA 谧列谧: 3 7

ATGAAATTCAACCCGCTGGACGAACTGGAAGAAACTCTGTACGAACA

GTTCACTTTCCAGCAGGGTGGTGGTGGTGGCGGTGGTAAGTTCAACCC

ACTGGATGAGCTGGAAGAGACTCTGTATGAACAGTTCACTTTCCAGCA

ACTCGAGGGTGGAGGCGGTGGGGACAAAACTCACACATGTCCACCTT

GCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTTTTCCTCTTCCCCCC

AAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATG

CGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTG

GTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGG

AGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCC

TGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCC

AACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAA

AGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGG

ATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCT

TCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCG

GAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCC

TTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAG

GGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC

TACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATAA -145- 1317281 Description of Invention Continued Page (141) Column Identification: No. 3 8

ATGGGTGCACAGCAGGAAGAATGCGAATGGGACCCATGGACTTGCGA

ACACATGCTCGAGGGTGGAGGCGGTGGGGACAAAACTCACACATGTC

CACCTTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTTTTCCTCTT

CCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGT

CACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTT

CAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGC

CGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTC

ACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAA

GGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAA

AGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCAT

CCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCA

AAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGG

CAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGAC

GGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGG

CAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCAC

AACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATAA 庠 column identification: No. 3 9

ATGCAGGAAGAATGCGAATGGGACCCATGGACTTGCGAACACATGGG

ATCCGGTTCTGCTACTGGTGGTTCCGGCTCCACCGCAAGCTCTGGTTCA

GGCAGTGCGACTCATCTCGAGGGTGGAGGCGGTGGGGACAAAACTCA

CACATGTCCACCTTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGT

TTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACC

CCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGA

GGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAA

GACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA

GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACA

AGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCA

TCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTG

CCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGC

CTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGC

AATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGA

CTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAG

CAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGC

TCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAA

ATAA -146- 1317281 Description of Invention Continued Page (142) Sequence Identification: No. 40.

ATGGGTGCACAGCAGGAAGAATGCGAATGGGACCCATGGACTTGCGA

ACACATGGGATCCGGTTCTGCTACTGGTGGTTCCGGCTCCACCGCAAG

CTCTGGTTCAGGCAGTGCGACTCATCAGGAAGAATGCGAATGGGACCC

ATGGACTTGCGAACACATGCTCGAGGGTGGAGGCGGTGGGGACAAAA

CTCACACATGTCCACCTTGCCCAGCACCTGAACTCCTGGGGGGACCGT

CAGTTTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCG

GACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACC

CTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATG

CCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTG

GTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAG

TACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAA

ACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACAC

CCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGAC

CTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGA

GAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGC

TGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACA

AGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATG

AGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGG

GTAAATAA queue identification: 4 1

ATGGACAAAACTCACACATGTCCACCTTGCCCAGCACCTGAACTCCTG

GGGGGACCGTCAGTTTTCCTCTTCCCCCCAAAACCCAAGGACACCCTC

ATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGC

CACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGA

GGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCA

CGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA

ATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCC

CCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACC

ACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCA

GGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGC

CGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCA

CGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCT

CACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTC

CGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTC

CCTGTCTCCGGGTAAAGGTGGAGGTGGTGGTGCACAGAAATTCAACCC

GCTGGACGAGCTGGAAGAGACTCTGTACGAACAGTTTACTTTTCAACA

GCTCGAGTAA • 147- Invention Description Continued Page 1317281 (143) Column Identification: 4 2

ATGGACAAAACTCACACATGTCCACCTTGCCCAGCACCTGAACTCCTG

GGGGGACCGTCAGTTTTCCTCTTCCCCCCAAAACCCAAGGACACCCTC

ATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGC

CACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGA

GGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCA

CGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA

ATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCC

CCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACC

ACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCA

GGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGC

CGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCA

CGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCT

CACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTC

CGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTC

CCTGTCTCCGGGTAAAGGTGGAGGTGGTGGTGCACAGGGATCCGGTTC

TGCTACTGGTGGTTCCGGCTCCACCGCAAGCTCTGGTTCAGGCAGTGC

GACTCATAAATTCAACCCGCTGGACGAACTGGAAGAAACTCTGTACGA

ACAGTTCACTTTCCAGCAACTCGAGTAA 识别 column identification: 4 3

ATGGACAAAACTCACACATGTCCACCTTGCCCAGCACCTGAACTCCTG

GGGGGACCGTCAGTTTTCCTCTTCCCCCCAAAACCCAAGGACACCCTC

ATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGC

CACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGA

GGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCA

CGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA

ATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCC

CCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACC

ACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCA

GGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGC

CGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACGA

CGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCT

CACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTC

CGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTC

CCTGTCTCCGGGTAAAGGTGGAGGTGGTGGTGCACAGAAATTCAACCC

GCTGGACGAAC Ding GGAAGAAACTC Ding GTACGAACAGTTCACTTTCCAGCA

GGGTGGTGGTGGTGGTGGCGGTGGTAAGTTCAACCCACTGGATGAGCT

GGAAGAGACTCTGTATGAACAGTTCACTTTCCAGCAACTCGAGTAA -148- 1317281 1 Invention Description Continued Buy (144) __--- Pain List Identification. 4 No. 4 -

ATGGACAAAACTCACACATGTCCACCTTGCCCAGCACCTGAACTCCTG

GGGGGACCGTC'AGTTTTCCTCTTCCCCCCAAAACCCAAGGACACCCTC

ATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGC

CACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGA

GGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCA

CGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA

ATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCC

CCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACC

ACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCA

GGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGC

CGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCA

CGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCT

CACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTC

CGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTC

CCTGTCTCCGGGTAAAGGTGGAGGTGGTGGTGCACAGCAGGAAGAAT

GCGAATGGGACCCATGGACTTGCGAACACATGCTCGAGTAA Year Identification·· 4 5

ATGGACAAAACTCACACATGTCCACCTTGCCCAGCACCTGAACTCCTG

GGGGGACCGTCAGTTTTCCTCTTCCCCCCAAAACCCAAGGACACCCTC

ATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGC

CACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGA

GGTGGATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCA

CGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA

ATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCC

CCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACC

ACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCA

GGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGC

CGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCA

CGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCT

CACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTC

CGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTC

CCTGTCTCCGGGTAAAGGTGGAGGTGGTGGTGCACAGGGATCCGGTTC

TGCTACTGGTGGTTCCGGCTCCACCGCAAGCTCTGGTTCAGGCAGTGC

GACTCATCAGGAAGAATGCGAATGGGACCCATGGACTTGCGAACACA

TGCTCGAGTAA -149- 1317281 Announcement of Invention (1) Column Identification: No. 46

ATGGACAAAACTCACACATGTCCACCTTGCCCAGCACCTGAACTCCTG

GGGGGACCGTCAGTTTTCCTCTTCCCCCCAAAACCCAAGGACACCCTC

ATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGC

CACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGA

GGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCA

CGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA

ATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCC

CCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACC

ACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCA

GGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGC

CGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCA

CGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCT

CACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTC

CGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTC

CCTGTCTCCGGGTAAAGGTGGAGGTGGTGGTGCACAGCAGGAAGAAT

GCGAATGGGACCCATGGACTTGCGAACACATGGGATCCGGTTCTGCTA

CTGGTGGTTCCGGCTCCACCGCAAGCTCTGGTTCAGGCAGCGCGACTC

ATCAGGAAGAATGCGAATGGGACCCATGGACTTGCGAACACATGCTC GAGTAA 庠 column identification: 4 7

ATGGGTGCACAGGAAGAATGCGAATGGGACCCATGGACTTGCGAACA

CATGGGTGGTGGTGGTGGTGGCGGTGGTAAATTCAACCCGCTGGACGA

ACTGGAAGAAACTCTGTACGAACAGTTCACTTTCCAGCAGGGATCCGG

TTCTGCTACTGGTGGTTCCGGCTCCACCGCAAGCTCTGGTTCAGGCAGT

GCGAGTCATCTCGAGGGTGGAGGCGGTGGgGACAAAACTCACACATGT

CCACCTTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTTTTCCTCT

TCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGG

TCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGT

TCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAG

CCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTC

ACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAA

GGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAA

AGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCAT

CCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCA

AAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGG

CAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGAC

GGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGG

CAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCAC

AACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATAA -150- 1317281 Description of Invention Continued Page (146) Sequence Identification: 48

ATGGACAAAACTCACACATGTCCACCTTGCCCAGCACCTGAACTCCTG

GGGGGACCGTCAGTTTTCCTCTTCCCCCCAAAACCCAAGGACACCCTC

ATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGC

CACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGA

GGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCA

CGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA

ATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCC

CCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACC

ACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCA

GGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGC

CGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCA

CGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCT

CACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTC

CGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTC

CCTGTCTCCGGGTAAAGGTGGAGGTGGTGGTGCACAGGGATCCGGTTC

TGCTACTGGTGGTTCCGGCTCCACCGCAAGCTCTGGTTCAGGCAGTGC

GACTCATAAATTCAACCCGCTGGACGAACTGGAAGAAACTCTGTACGA

ACAGTTCACTTTCCAGCAGGGTGGTGGCGGTGGTCAGGAAGAATGCGA

ATGGGACCCATGGACTTGCGAACACATGCTCGAGTAA Sequence identification: No. 4 9 .

ATGGGTGCACAGTTCGACTACTGCGAAGGTGTTGAAGACCCGTTCACT

TTCGGTTGCGACAACCACCTCGAGGGTGGAGGCGGTGGGGACAAAAC

TCACACATGTCCACCTTGCCCAGCACCTGAACTCCTGGGGGGACCGTC

AGTTTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGG

ACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCT

GAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCC

AAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGT

CAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTA

CAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAAC

CATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCC

TGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCT

GCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGA

GCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTG

GACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAG

AGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAG

GCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT

AAATAA -151 - 1317281 _ (147) Description of the invention Title column identification: No. 5 0

ATGGGTGCACAGCAGTACGGTTGCGACGGTTTTCTGTACGGTTGCATG

ATCAACCTCGAGGGTGGAGGCGGTGGGGACAAAACTCACACATGTCC

ACCTTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTTTTCCTCTTC

CCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTC

ACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTC

AACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCC

GCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCA

CCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG

GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAA

GCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCC

CGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAA

AGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGC

AGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGAC

GGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGG

CAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCAC

AACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATAA 庠 column identification: 5 1

ATGGGTGCACAGAAACGCCCATGCGAAGAAATGTGGGGTGGTTGCAA

CTACGACCTCGAGGGTGGAGGCGGTGGGGACAAAACTCACACATGTC

CACCTTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTTTTCCTCTT

CCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGT

CACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTT

CAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGC

CGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTC

ACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAA

GGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAA

AGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCAT

CCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCA

AAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGG

CAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGAC

GGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGG

CAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCAC

AACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATAA -152- 1317281 Announcement of Invention (1) Column Identification: 5 2

ATGGGTGCACAGCACCAGATCTGCAAATGGGACCCGTGGACCTGCAA

ACACTGGCTCGAGGGTGGAGGCGGTGGGGACAAAACTCACACATGTC

CACCTTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTTTTCCTCTT

CCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGT

CACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTT

CAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGC

CGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTC

ACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAA

GGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAA

AGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCAT

CCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCA

AAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGG

CAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGAC

GGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGG

CAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCAC

AACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATAA 庠 column identification: 5 3

ATGGGTGCACAGAAACGTCCATGCGAAGAAATCTTCGGTGGTTGCACC

TACCAGCTCGAGGGTGGAGGCGGTGGGGACAAAACTCACACATGTCC

ACCTTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTTTTCCTCTTC

CCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTC

ACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTC

AACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCC

GCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCA

CCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAG

GTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAA

GCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCC

CGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAA

AGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGC

AGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGAC

GGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGG

CAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCAC

AACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATAA -153 - 1317281 DESCRIPTION OF THE INVENTION continued (149) Example 6 Peptibody assay Four peptibodies were tested using a neutralization ELISA and three peptibodies were tested using an affinity ELISA. The results are stated in Table 5.

Table 5 hAng-2 mAng-2 hAng-1 Peptide IC 50 (nM) EC 50 (nM) IC 50 (nM) EC 50 (nM) IC 50 (nM) EC 50 (nM) 2xCon4 (C) 1K 0.04 0.02 Con4-Ll(C) 0.05 0.04 Con4 (C) 0.20 0.30 2xLl (N) 0.65 0.80 Con4 (N) 0.85 0.03 0.72 0.07 No inhibition No binding 2xLl (C) 0.90 1.0 Con4 (N) 1K-WT 1.9 LI ( N) 6 11 No inhibition C17 (N) 9 13 No inhibition 12-9 (N) 21 7.7 No inhibition Coni (N) 26 ~ 200 No inhibition 8-14 (N) 45 33 No inhibition LI ( C) 65 37 8-8 (N) 80 ~700 no inhibition negative control peptibody 4883 no inhibition no binding no inhibition no binding no inhibition no binding negative control peptide 4883 amino acid The sequence is as follows (added in the Fc section -154-1317281 (150). The description of the continuation line is marked with the link ""GGGGG" and the peptide portion is in bold):

MDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCWVD

VSHEDPEVKJNWYVDGVEVHNAKTKPREEOYNSTYRVVSVLTVL

HODWLNGKEYKCKVSNKALPAPIEKTISKAKGOPREPOVYTLPPS

RDELTKNOVSLTCLVKGFYPSDIAVEWESNGOPENNYKTTPPVLD

SDGSFFLYSKLTVDKSRWOOGNVFSCSVMHEALHNHYTOKSLSLS PGK-GGGGG-CTAGYHWNSDCECCRRN (Sequence Identification: No. 243)

It should be understood that the use of "no inhibition" is not intended to mean that the compound has no inhibitory properties. The term "non-inhibiting" as used herein, rather means that those compounds exhibit a greater IC50 value than 1000 nM under the conditions described herein, in use and in an ELISA assay, which is a screening The highest concentration of these compounds. Although no significant inhibitory properties were observed for the molecules labeled "no inhibition", it should be understood that in fact, under different assay conditions' or in different assays' The inhibitory properties of those molecules are confirmed. In a preferred embodiment, it is understood that the present invention relates to peptibodies having inhibitory properties when using the assays described herein. Testing of two peptibodies using an affinity BIAcore assay (like The results are described in Example 2. The results are set forth in Table 6 below. Table 6 Peptide fPh) #hAng-2 and mAng-2 Affinity. Force. hAng-2 hAng-2 Peptide KD (nM) ka ( 1/Ms) kd (1/s) KD (nM) ka (1/Ms) kd (1/s) Pb LI (N) 3.1 2.9 x 105 9.1 x 10-4 0.42 5.6 x 105 2.3 x 10-4 Con4 (N) 0.67 3.3 x 105 2.2 x ισ4 0.60 7.3 x 105 4.4 x 10"4 TN12-9 (N) 8.2 1.2 x 105 1.0 x 10'3 0.32 7.2 x 105 2.3 x 10"4 -155- 1317281 (151) Description of the Invention Continued Page 1 Example 7

The efficacy of systemic administration of Ang-2 peptone was studied by subcutaneous administration of Ang-2 peptone, TN8-Con4-C, 72 hours after tumor challenge. The doses of the peptibodies used were 1000, 200, 40 and 8 μg/mouse/day. All animals were given a total of 20 doses. Tumor volume and body weight were recorded three times a week. At the end of the study, the animals were sacrificed and their serum was collected and the peptone content was measured by ELISA. Tumors and normal tissues in the list were collected from all groups. The result is shown in Figure 1. As can be seen, a significant difference in tumor growth was observed between the Ang-2 peptidial treatment group and the vehicle control group. All four doses of Ang-2 peptibodies inhibited tumor growth compared to the vehicle control group (p<0.0001 versus vehicle control, using repeated measures ANOVA). In contrast, tumors in the control group continued to grow at a much higher rate. Treatment with peptibody did not have a significant effect on the final body weight, organ weight or hematological parameters of the animals treated at the above doses. Example 8

1. Construction of Ang-2 secondary peptide Kang A. Electroreactive potential of E. coli cells Purchased Epicurian Coli® XL 1-Blue MRF electroporation potential cells from Stratagene (Stratagene Cloning Systems, La Jolla, CA) (Stratagene # 200158). B. Strip modification of dCESI vector PCR was performed using an extended-length template PCR system (Roche Diagnostics Corp., Indianapolis, IN) using 1 μg of pCES1 vector (TargetQuest Inc.) as template-156-1317281 (152). . The PCR mixture has a volume of 100 μl, which contains lx PCR buffer solution, two primers: 5, -CAAACGAATGGATCCTCATTAAAGCCAGA_3, (sequence recognition: No. 244) and 5,-GGTGGTGCGGCCGCACTCGAGACTGTTGAAAGTTGTTTAGCA-3, (sequence identification: No. 245) each 200 nM, 200 nM dNTPs and 3 units (U) of Tag DNA polymerase. The TRIO-Thermoblock (Biometra) PCR system was performed as follows: 94 ° C for 5 minutes; 94 ° C for 30 seconds, 50 ° C for 30 seconds, 72 ° C for 45 seconds for 30 rounds: and 72 ° C for 10 minutes; cooling to 4 ° C.

The P C R product was then run on a 1% agarose gel and purified using a QIAGEN spin column (QIAGEN Inc., Valencia, CA) according to the manufacturer's draft. Using 5 μl of PCR product, and two primers, 5匕CAAACGAATGGATCCTCATTAAAGCCAGA-3, (SEQ ID NO: 246) and 5I-AACACAAAAGTGCACAGGGTGGAGGTGGTGGTGCGGCCGCACT-3, (SEQ ID NO: 247) each 200 nM under the same PCR conditions as above , performing a second PCR reaction.

The PCR product and the original pCES1 vector were then separately digested in a 100 μl reaction containing 1 χΝΕΒ 2 buffer solution, 60 units of ApaLI (New England Biolabs, Beverly, ΜΑ), 60 units of BamHI (New England Biolabs) at 37 °C. hour. The digested DNA was then purified using a QIAGEN spin column and incubated overnight at room temperature in a 40 microliter reaction containing lx ligation buffer and 40 units of T4 DNA ligase (New England Biolabs). together. The vector was transferred into E. coli and incubated overnight at 37 °C. A single colony that was isolated was selected and the plastid was purified using a QIAGEN spin column. Confirming the correct insert by DNA sequencing -157· Description of invention Continued: 1317281 (153)

C. Preparation of vector DNA 1 microgram of modified pCES1 vector DNA (obtained from paragraph 1B above) was converted to 2 microvolts, 25 microfarads and 200 ohms of Gene Pulser II (BIO-RAD, Hercules, CA). 40 microliters of electroreactive potential of XLl-blue E. coli (from paragraph 1A above). The transformed bacterial sample was then immediately transferred to contain 960 microliters of SOC (2% trypsin hydrolyzed enzyme, 0.5% yeast extract, 10 mM NaCn, 2.5 mM KC1,

In a test tube of 20 mM glucose, 10 mM MgS 4 , 10 mM MgCh), the culture was allowed to grow at 37 ° C and shaken for 1 hour. The cells were then spread on 2 x YTAGT (2xYT, containing 100 μg/ml ampicillin, 12.5 μg/ml tetracycline and 2% glucose) on agar plates and incubated overnight at 37 ° C. Colonies were used and inoculated with 2 liters of 2x YTAGT medium and shaken overnight at 37 °C. Purify the plastid vector DNA using the QIAGEN plastid Maxi kit according to the manufacturer's draft »

D. Digestion of vector DNA

A total of approximately 2000 μg of carrier DNA (from paragraph ic above) was digested overnight at 37 ° C in a 5000 μl reaction containing 2,NEB buffer solution of 2,300 units of ApaLI and 300 units of Xhol. The restriction digestion reaction was incubated at 37 ° C overnight and analyzed in a pre-made 0.8% agarose gel (Embi Tec, San Diego, CA). The linearized vector DNA was then excised from the gel and extracted with a QIAquick Gel Extraction Kit (QIAGEN Inc.) according to the manufacturer's instructions. E. Borrowing of ruthenium nucleotides - 158- 1317281 (154) Continuation of the page Designing six library oligonucleotides (1 fixed and 5 anesthetic) based on sequences derived from the above results . A fixed library oligonucleotide is: 5'-CACAGTGCACAGGGTNNKNNKNNKNNKNNKNNKNNKS ARTGGGATCCGTGGASCNNKNNKNNKNNKNNKNNKNNKCATT CTCTCGAGATCA-3' (Library 20) (sequence identification |J: 248) and two 70% anesthetized library oligonucleotides are as follows:

5'-CACAGTGCACAGGGTNNKNNKNNKaaKcgKccKNNKga KgaKatKttKggKNNKacKtaKcaKNNKNNKNNKCATTCTC TCGAGATCA-3, (library number 27) (SEQ ID: No. 249) 5'-CACAGTGCACAGGGTNNKaaKttKaaKccKctKgaKgaKctKgaKga KacKctKtaKgaKcaKttKacKttKcaKcaKNNKCATTCTCTCGAGATCA-3 '(library number 99) (SEQ ID: 250);

Lower case letters represent a mixture of 70% of the specified bases and 10% of each of the other three nucleotides. 91% of the other three oligonucleotide library anesthetized drill acid following: 5'-CACAGTGCACAGGGTNNKNNKNNKcaKgaKgaKTGCgaKtg KgaKccKtgKacKTGCgaKcaKatKNNKNNKNNKCATTCTCTCGAGA TCA-3, (library number 94) (SEQ ID: No. 251); 5'-CACAGTGCACAGGGTNNKttKgaKtaKNNKgaKggKgtKgaKgaKcc KttKacKttKggKNNKgaKaaKcaKNNKCATTCTCTCGAGATCA-3 '(library number 25) ( Sequence identification: No. 252); and 5'-CACAGTGCACAGGGTNNKaaKttKaKccKctKgaKgaKctKgaKga KacKctKtaKgaKcaKttKacKttKcaKcaKNNKCATTCTCTCGAGATCA-3' (Library No. 26) (sequence identification: No. 253); -159- 1317281 (155) Description of the continuation on the above oligo, familiar with this artist should Understanding "N" means that each of the four nucleotides (A, T, C, and G) is equally represented during oligo synthesis, and "K" indicates equal representation of nucleotides during oligo synthesis 〇 and τ. Lowercase letters represent a mixture of 91% of the specified bases and 3% of the other three cores: each of the acids. These oligonucleotides are used as templates in PCR. For PCR reactions, PCR using extended high fidelity System kit (R0Che Diagnostics Corp.). Contains InM library Diclofenac, 1 x PCR buffer, primers:

5,-CACAGTGCACAGGGT-3, (sequence recognition: No. 254); and 5'-TGATCTCGAGAGAATG-3 (sequence recognition: No. 255);

Each of the library was expanded in a 96-well 50 μl PCR reaction with 300 μM each; 200 μΜ dNTP, 1.5 mM MgCl 2 and 350 units of extended polymerase. The following procedure was performed using a thermal cycler (GeneAmp PCR System 9700, Applied Biosystems): 94 ° C for 5 minutes; (94 ° C for 30 seconds, 52.5 ° C for 60 seconds, 72 ° C for 30 seconds) X 25 rounds; 72 ° C 10 minutes; cooled to 4 °C. The QIAquick PCR Purification Kit (QIAGEN Inc. Catalog #28104) was then used to remove free nucleotides according to the manufacturer's draft. E. Digestive nucleus decanoic acid The PCR product of each pool (paragraph 1E) was digested overnight at 37 ° C in a 1200 microliter reaction containing 2, 750 units of ApaLI and 750 units of Xhol in 2x NEB buffer solution. The digested DNA was separated on a pre-made 3% agarose gel (Embi Tec). The DNA strand of interest obtained from each reaction was excised from the gel and subjected to COSTAR spin-X centrifuge tube filter paper, 0.22 μm acetic acid-160-1317281 _ (156) Description of the Invention Continued Cellulose (Corning Inc ., catalog #8160) extraction. G. The carrier is stunned and the acid is connected.

450 μl of ligation reaction was carried out overnight at 16 ° C, containing a linearized vector at a molar ratio of 1:5 (paragraph 1D) and each digested library PCR product (paragraph lF), lxNEB ligation buffer solution And 20,000 units of T4 DNA ligase. The ligation product was incubated at 65 °C for 20 minutes to inactivate the T4 DNA ligase and incubated with 100 units of Notl for 2 hours at 37 °C to minimize self-ligation of the vector. The ligation product was then purified by standard phenol/chloroform extraction - (Molecular Cloning: A Laboratory Manual, Maniatis et al., 3rd edition, Cold Spring Harbor Laboratory Press, 2000) and resuspended in 120 microliters of H20. H. Electrical penetration of Koreanization

For each library, 12 electroporation reactions were performed. For each transformation, mix 10 μl of ligation of the carrier DNA (paragraph 1G) and 300 μl of XL1-BLUE MRF' cells (paragraph 1A) in a 0.2-cm cuvette (BIO-RAD). The resulting mixture was pulsed by a Gene Pulser(R) set to 2500 volts, 25 microfarads and 200 ohms. The transformed bacteria obtained from the 12 electroporation reactions were then mixed and transferred to a flask containing 26 ml of SOC and cultured at 37 ° C for 1 hour. The cells were added to 450 ml of 2xYTAG and allowed to grow at 37 ° C for 5 hours with shaking. The cells were centrifuged at 4000 rpm for 15 minutes at 4 °C. The cell pellet was then resuspended in 12 ml of 15% glycerol/2xYT and stored at -8 (TC). This is the original mother liquor of the library. The titration shows that the size of the library is 5·0χ109 (Library 20), 3_3xl01 () (Library 94), 4.7xl09 (Library 25), 5.0xl09 (Library 26), 3.0X109 (Library 27), and 4·2χ109 (Library 99) Single-161 - 1317281 __ (157 The invention describes the continuation of the transformant. 2. The expansion of the library A. The manufacture of secondary clean mother liquor from the original stock cell mother liquor (from paragraph 1H above), using a library size including 10X excess Sufficient cells to inoculate 2xYTAGT (2YT with 100 μg/ml ampicillin, 12.5 μg/ml tetracycline and 2% glucose) to give a starting OD6QG of 0.1. Allow the culture to grow at 37 ° C, and Shake for a few hours until OD6 〇〇 = 0.5. One tenth of an aliquot was taken from each pool and grown in a different flask for an additional 2 hours at 37 ° C. Then at 4 ° C, Using a Beckman JA-14 rotator, centrifuge at 4000 rpm for 10 minutes, and resuspend the bacterial pellets at 7.0 mL (each ) 15% glycerol/2xYT, stored at -80 ° C. B. Phage induction Add the M13K07 helper phage aliquot (Amersham Pharmacia Biotech) to the remaining bacterial culture with OD6 () () = 0.5 (from paragraph 2A above), to a final concentration of 3xl09 pfu/ml. Allow the helper phage to infect the bacteria for 30 minutes at 37 ° C, without shaking up, and then slowly vortex for 30 minutes. The infected cells were centrifuged at 5000 rpm for 15 minutes at ° C. The cell pellet was resuspended in the same volume (from paragraph 2A above) in 2x YTAK medium (with 100 μg/ml ampicillin and 40 μg) /ml of kampromycin in 2YT) Allows the production of phage plastids to occur overnight at 30 ° C, while being shaken up. C. The phage phage is centrifuged at 5000 rpm from 4 ° C. Bacterial Culture of Paragraph 2B 1317281 Description of the Invention (158)

15 minutes. The supernatant was then transferred to a new bottle and 0.2 volume of 20% PEG/2.5 M NaCl was added and incubated on ice for 1 hour to precipitate the phage. The precipitated phage plastids were centrifuged at 10,000 rpm for 30 minutes at 4 ° C and carefully resuspended in 100 ml of ice-cold PBS. The remaining cells were removed by centrifugation at 4000 rpm for 10 minutes at 4 ° C to further purify the phage plastid solution by adding 0.2 volumes of 20% PEG/2.5M.

NaCl, which precipitates phage plastids. Centrifuge at 10,000 rpm at 4 ° C

The plastids were incubated for 30 minutes and the phage plastid pellets were resuspended in 18 ml of ice-cold PBS. 6 ml of a 60% glycerol solution was added to the phage plastid solution and stored at -80 °C. The valence of phage plastids was determined by standard procedures (Molecular Cloning, Maniatis et al., 3rd edition). 3. Select Ang-2 binding phage A. Fix Ang-2 on magnetic beads

Biotinylated Ang-2 (from paragraph 3A above) was immobilized on Dynabead M-280 Streptomyces antibiotics at a concentration of 2000 micrograms of Ang-2 protein per 100 milliliters of the mother's bead mother liquor. Protein (DYNAL, Lake Success, NY). After the beads were attracted to one side of the tube using a magnet and the liquid was aspirated, the beads were washed twice with phosphate buffered saline (PBS) and resuspended in PBS. Biotinylated Ang-2 protein was added to the washed beads at the above concentrations and incubated at room temperature for 1 hour. The Ang-2 coated beads were then blocked by the addition of BSA to a final concentration of 2% and incubated overnight at 4 °C and spun. The resulting Ang-2 coated beads were then washed twice with PBST (PBS with 0.05% Tween-20) before accepting the selection procedure. -163- 1317281 _ (159) Description of the Invention Continued 彳 B. Use the coated Ang-2 beads to select

Approximately 1000-fold pool equivalent of phage plastids (from paragraph 2C above) was blocked using 1 ml of PBS containing 2% BSA for 1 hour. This was added to the blank beads (the same beads as in paragraph 3A but not coated with Ang-2 protein) and the mixture was incubated for 15 minutes at room temperature and spun to allow blocked phage The plastid sample received three negative selection steps. The phage plastid containing the supernatant was aspirated using a magnet and transferred to a second blank containing beads (the same beads as described in paragraph 3A above, but without the coated-Ang-2 protein) The mixture was incubated in a test tube at room temperature for 15 minutes and rotated.

Repeat the program. The phage plastid containing the supernatant was aspirated using a magnet and transferred to a new tube containing the coated Ang-2 protein beads (obtained from paragraph 3A) and the mixture was incubated for 1 hour at room temperature and rotated. . After discarding the supernatant, the beads bound to the phage plastids were washed 10 times with 2% milk-PBS; 10 times with 2% BSA-PBS; 10 times with PBST; and washed twice with PBS. The phage plastids were allowed to elute on a rotator in 1 ml of 100 mM triethylamine solution (Sigma, St. Louis, MO) for 10 minutes. The pH of the solution containing the phage plastid was neutralized by adding 0.5 ml of 1 PCT 1 ^ -11 (: 1 117.5). Using the obtained phage plastid, 10 ml of newly grown XL1-Blue MRF· bacteria (OD6 〇〇 about 0.5) was infected at 37 ° C for 30 minutes, and shaking was continued for 30 minutes without shaking. All infected XL1-BLUE MRF' cells were then plated on a 15 x 15 cm 2x YTAG pan and incubated overnight at 30 °C. C. Induction and contraction of phage -164- 1317281 _ (160) I invention description continuation page

A 10 ml aliquot of 2x YTAGT medium was added to the plate (from paragraph 3B) and the XL1-BLUE MRF' cells were resuspended. All XL1-BLUE MRF' cells were collected in tubes and 250 microliter aliquots of these cells were added to 25 ml 2xYTAGT and grown at 37 °C until OD6 〇〇 = 0.5. M13K07 helper phage was added to a final concentration of 3 x 109 pfu/ml and incubated at 37 ° C for 30 minutes without shaking, and then slowly shaken for 30 minutes. The cells were centrifuged at 5000 rpm for 10 minutes at 4 ° C and resuspended in 25 ml of 2x YTAK. These bacteria were allowed to grow overnight at 30 ° C and shaken. The induced phage plastids were harvested and purified according to paragraph 2C. D. Round 2 selection

The selection of the second round, as outlined in paragraphs 3B to 3C, is except for the following. Approximately 100-fold pool equivalent phage plastids from paragraph 3C were used as input phage plastids. The content of biotinylated Ang-2 protein (paragraph 3A) coated on Dynabead M-280 Streptomyces avidin was reduced to 20 nanograms. The beads bound to the phage plastids were then washed 10 times with 2% milk-PBS; 10 times with 2% BSA-PBS; 10 times with PBST, where the final rinse involved at room temperature in PBST Cultivate for 60 minutes. The beads were rinsed twice with PBS. The elution conditions were the same as in the first round (segment 3B). E. Round 3 Choice The selection of the third round is outlined in paragraphs 3B to 3C, except for the following. Approximately 10-fold pool equivalent phage plastid from paragraph 3D was used as the input phage plastid. Approximately 20 nanograms of biotinylated Ang-2 protein (available from paragraph 3A) was used to coat Dynabead M-280-165-1317281 (161) Illustrated Continued, Streptomyces avidin. The beads bound to the phage plastids were washed 10 times with 2% milk-PBS; 10 times with 2% BSA-PBS; 10 times with PBST, where the final rinse involved incubation in PBST at room temperature 60 minutes. The beads were rinsed twice with PBS. The elution conditions were the same as in the first round (paragraph 3B). F. Round 4 Selection The selection of the fourth round was carried out in accordance with paragraphs 3B to 3C, except for the following. The phage plastid from the library equivalent of paragraph 3E was used as the input phage plastid. For pools 25, 26 and 27, the amount of biotinylated Ang-2 protein (paragraph 3A) coated on Dynabead M-280 Streptomyces avidin was reduced to 0.4 nanograms. For pools 20 and 94, the coating content was maintained at 2 nanograms for the third round. Library 99 does not perform the selection step of the fourth round. The elution conditions were the same as in the first round (paragraph 3B). 4. Selection analysis A. Preparation of the main culture tray A single colony selected from the second round was selected and seeded in a 96-well culture dish containing 120 μl of 2x YTAGT per well. The 96-well culture plate was incubated overnight in a 30 ° C shaker. 40 microliters of 60% glycerol was added to each well and stored at -80 °C.

B. Phage plastid ELISA Approximately 2 microliter aliquots of cells from the primary culture plate (obtained from paragraph 4A above) were seeded in fresh Costar® 96-well plates (Corning incorporated, Corning, NY, catalog) In #9794), it contained 100 μl of 2xYTAGT per well, and the cells of the new plate were grown at 37 ° C until about OD6 〇〇 = 〇 5. 1317281 Description of the Invention (162)

40 μl of 2xYTAGT containing M13K07 helper phage (1·5 X 1013> fii ml) was added to each well, and the 96-well culture plate was incubated at 37 ° C for 30 minutes without shaking, and then slowly oscillated. 30 minutes. The plates were centrifuged at 2000 rpm (Beckman CS-6R tabletop centrifuge) for 10 minutes at 4 °C. The supernatant was removed from the wells and each cell pellet was resuspended using 150 microliters of 2x YTAK per well. For phage plastid expression, the plate was incubated overnight at 30 °C.

Human Ang-2 protein was plated on a 96-well Maxisorp plate (NUNC) overnight at 1 μg/ml in lxPBS at 4 °C. On the next day, the cell culture was centrifuged at 2000 rpm for 10 minutes at 4 °C. Ten microliters of the supernatant from each well was transferred to a new 96-well plate containing BSA/PBS solution, and the supernatant was diluted 1:10. The resulting mixture was incubated at room temperature for 1 hour and shaken to block phage plastids. At this time, block coating at 400 μL of 2% BSA/PBS solution per well at room temperature

The plate of Ang-2 protein was incubated for 1 hour while shaking. The BSA solution was discarded and the wells were rinsed 3 times with PBS solution. In the last rinse step

Thereafter, 100 μl of the blocked phage plastid solution was added to each well of the culture plate coated with Ang-2 protein and the control plate, and incubated at room temperature for 1 hour, and stirred. The liquid was discarded and the wells were washed three times with PBST solution. 100 μl of HRP-conjugated anti-M13 mAb (Amersham Pharmacia Biotech) was added to each well of the Ang-2 protein and control culture plates at a dilution of 15,000, and these cultures were cultured at room temperature. Plate for 1 hour and stir. The liquid was again discarded and the wells were rinsed three times with PBST solution. 100 microliters of LumiGLO chemiluminescent substrate (Kirkegaard & Perry Laboratories, Gaithersburg, MD) was added to the wells, and by -167- invention description continued 丨 1317281 (163)

The holes were read on a Luminoskan Ascent DLRearly machine (Labsystems, Franklin, MA). C. Sequencing of the phagetric pure germ line 1 microliter of bacteria obtained from each well of the main culture plate (paragraph 4A) was used as a template. Each PCR mixture has a volume of 50 μl, which contains a 1 X PCR buffer solution, two primers: S'-GTTAGCTCACTCATTAGGCAd (sequence recognition: No. 256) and 5,-GTACCGTAACACTGAGTTTCG-3, (sequence identification: No. 257);

300 nM each; 200 uM dNTP, 2 mM MgCl2 and 2.5 units of taq DNA polymerase (Roche Molecular Biochemicals). Using the GeneAmp PCR System 9700 (Applied Biosystems), the following procedure was performed: 94°c for 5 minutes; (94°C for 45 seconds '55°C for 45 seconds, 72°C for 90 seconds) for 40 rounds; 72°C for 10 minutes ; Cool to 4 ° C. The PCR product was purified using a QIAquick 96 PCR purification kit (QIAGEN Inc.) according to the manufacturer's instructions. Using the primer 5, -TTACACTTTATGCTTCCG-3, (sequence recognition: No. 258)', the sequence of all purified PCR products was determined using the aBI 3770 sequencer (Perkin Elmer) according to the manufacturer's instructions. 5. Sequencer column F will have the peptide sequence that has been translated from the nucleotide sequence (obtained from paragraph 4 above, consistent with the data of £1^8), which is considered to be shown in the wells coated with 八一^2 High OD readings and pure lines showing low OD readings in wells coated with BSA are more important. It is also considered important to have multiple sequences. 24 peptide sequences were selected from pool 20, from library 94. 26 peptide sequences were selected, 7 peptide sequences were selected from library 25, 18 peptide sequences were selected from library 26, 6 peptide sequences were selected from library 27, and 4 peptide sequences were selected from library 99 for further -168- 1317281 _ (164) Description of the invention Continued page analysis and peptidomimetic production. In addition, 11 consistent sequences were also derived from pools 20 and 94, and 3 consensuses were derived from pools 26 and 99. Sequences, and two consensus sequences were deduced from library 25 for the production of peptibodies. The peptibodies in Table 7 were evaluated using the neutralized ELISA draft described in Example 10 herein. The results. Table 7 Co4-derived Affinity-Mature Pbs hAng-2: Tie2 IC5〇(nM) Peptide Sequence (SEQ ID NO:) Con4-44 (C) 0.09 M-Fc-G GGGGAQ-PIRQEECDWDPWTCEHMWEV-LE (SEQ ID NO: 259) Con4-40 (C) 0.10 M-Fc-GGGGGAQ-TNIQEECEWDPWTCDHMPGK-LE (Sequence recognition: No. 260) Con4-4 (C) 0.12 M-Fc-GGGGGAQ-WYEQDACEWDPWTCEHMAEV- LE (SEQ ID NO: 261) Con4-31 (C) 0.16 M-Fc-GGGGGAQ-MRLQEVCEWDPWTCEHMENV-LE (Sequence recognition: No. 262) Con4-C5 (C) 0.16 M-Fc-GGGGGAQ-AATQEECEWDPWTCEHMPRS-LE (Sequence recognition :263) Con4-42 (C) 0.17 M-Fc-GGGGGAQ-LRHQEGCEWDPWTCEHMFDW-LE (Sequence recognition: No. 264)

-169- 1317281 _ (165) I Invention Description Continued

Con4-35 (C) 0.18 M-Fc-GGGGGAQ-VPRQKDCEWDPWTCEHMYVG-LE (SEQ ID NO: 265) Con4-43 (C) 0.18 M-Fc-GGGGGAQ: SISHEECEWDPWTCEHMQVG-LE (Sequence Recognition: No. 266) Con4-49 ( C) 0.19 M-Fc-GGGGGAQ-WAAQEECEWDPWTCEHMGRM-LE (SEQ ID NO: 267) Con4-27 (C) 0.22 M-Fc-GGGGGAQ-TWPQDKCEWDPWTCEHMGST-LE (Sequence recognition: No. 268) Con4-48 (C) 0.26 M -Fc-GGGGGAQ-GHSQEECGWDPWTCEHMGTS-LE (SEQ ID NO: 269) Con4-46 (C) 0.26 M-Fc-GGGGGAQ-QHWQEECEWDPWTCDHMPSK-LE (Sequence Recognition·· No. 270) Con4-41 (C) 0.26 M-Fc- GGGGGAQ- — NVRQEKCEWDPWTCEHMPVR-LE (SEQ ID NO: 271) Con4-36 (C) 0.28 M-Fc-GGGGGAQ-KSGQVECNWDPWTCEHMPRN-LE (Sequence recognition: No. 272) Con4-34 (C) 0.28 M-Fc-GGGGGAQ- VKTQEHCDWDPWTCEHMREW -LE (SEQ ID NO: 273) Con4-28 (C) 0.30 M-Fc-GGGGGAQ- AWGQEGCDWDPWTCEHMLPM-LE (Sequence recognition: No. 274) Con4-39 (C) 0.30 M-Fc-GGGGGAQ-PVNQEDCEWDPWTCEHMPPM-LE (sequence Identification: No. 275) Con4-25 (C) 0.31 M-Fc-GGGGGAQ-RAPQEDCEWDPWTCAHMDIK-LE (Sequence Identification: No. 276)

-170- 1317281 _ (166) Description of the Invention Continued

Con4-50 (C) 0.38 M-Fc-GGGGGAQ-HGQNMECEWDPWTCEHMFRY-LE (SEQ ID NO: 277) Con4-38 (C) 0.40 M-Fc-GGGGGAQ-PRLQEECVWDPWTCEHMPLR-LE (Sequence Recognition·· No. 278) Con4-29 (C) 0.41 M-Fc-GGGGGAQ-RTTOEKCEWDPWTCEHMESQ-LE (SEQ ID NO: 279) Con4-47 (C) 0.44 M-Fc-GGGGGAQ-QTSQEDCVWDPWTCDHMVSS-LE (Sequence recognition: No. 280) Con4-20 (C) 0.48 M-Fc-GGGGGAQ-QVIGRPCEWDPWTCEHLEGL-LE (SEQ ID NO: 281) Con4-45 (C) 0.48 M-Fc-GGGGGAQ- WAOOEECAWDPWTCDHMYGL-LE (SEQ ID NO: 282) Con4-37 (C) 0.49 M-Fc- GGGGGAQ-T .PGOEDCEWDP WTCEHMVRS-LE (Sequence recognition: No. 283) Con4-33 (C) 0.52 M-Fc-GGGGGAQ-PMNQVECDWDPWTCEHMPRS-LE (Sequence recognition: No. 284) AC2-Con4 (C) 0.52 M-Fc-GGGGGAQ -FGWSHGCEWDPWTCEHMGST-LE (SEQ ID NO: 285) Con4-32 (C) 0.75 M-Fc-GGGGGAQ-KSTQDDCDWDPWTCEHMVGP-LE (SEQ ID NO: 286) Con4-17(C) 0.96 M-Fc-GGGGGAQ- ' GPRISTCQWDPWTCEHMDQL- LE (Sequence recognition: No. 287) Con4-8 (C) 1.20 M-Fc-GGGGGAQ-STIGDMCEWDPWTCAHMQVD-LE (Sequence recognition: No. 288) AC4-Con4 (C) 1.54 M- Fc-GGGGGAQ-VLGGQGCEWDPWTCRLLQGW-LE (SEQ ID NO: 289) Con4-l (C) 2.47 M-Fc-GGGGGAQ-VLGGQGCQWDPWTCSHLEDG-LE (Sequence recognition: No. 290) Con4-Cl (C) 2.75 M-Fc-GGGGGAQ- TTIGSMCEWDPWTCAHMQGG-LE -171 - 1317281 _ (167) Description of the Invention (Sequence Identification: No. 291) Con4-21 (C) 3.21 M-Fc-GGGGGAQ-TKGKSVCQWDPWTCSHMQSG-LE (Sequence Identification: No. 292) Con4-C2 (C) 3.75 M-Fc-GGGGGAQ-TTIGSMCQWDPWTCAHMQGG-LE (SEQ ID NO: 293) Con4-18(C) 4.80 M-Fc-GGGGGAQ- WVNEWCEWDPWTCNHWDTP-LE (Sequence recognition: No. 294) Con4-19 (C) 5.76 M- Fc-GGGGGAQ- VVQVGMCQWDPWTCKHMRLQ-LE (SEQ ID NO: 295) Con4-16(C) 6.94 M-Fc-GGGGGAQ-AVGSQTCEWDPWTCAHLVEV-LE (Sequence recognition: No. 296) Con4-ll (C) 9.70 M-Fc-GGGGGAQ- QGMKMFCEWDPWTCAHIVYR-LE (SEQ ID NO: 297) Con4-C4 (C) 9.80 M-Fc-GGGGGAQ-TTIGSMCQWDPWTCEHMQGG-LE (Sequence recognition: No. 298) Con4-23 (C) 9.88 M-Fc-GGGGGAQ-TSQRVGCEWDPWTCQHLTYT-LE ( Sequence Identification: No. 299) Con4-15 (C) 15.00 M-Fc-GGGGGAQ-QWSWPPCEWDPWTCQTVWPS-LE (Sequence Identification: No. 300) Con4- 9 (C) 20.11 M-Fc-GGGGGAQ-GTSPSFCQWDPWTCSHMVQG-LE (Sequence Recognition·· No. 301) . Con4-10 (C) 86.61 M-Fc-GGGGGAQ-TQGLHQCEWDPWTCKVLWPS-LE (Sequence Identification: No. 302) Con4-22 ( C) 150.00 M-Fc-GGGGGAQ- VWRSQVCQWDPWTCNLGGDW-LE (sequence recognition: No. 303) Con4-3 (C) 281.50 M-Fc-GGGGGAQ-DKILEECQWDPWTCQFFYGA-LE (sequence identification: 304)

-172- 1317281 (168) Description of the Invention Continued

Con4-5 (C) No inhibition M-Fc-GGGGGAQ-ATFARQCQWDPWTCALGGNW-LE (Sequence recognition: No. 305) Con4-30 (C) No inhibition M-Fc-GGGGGAQ-GPAQEECEWDPWTCEPLPLM-LE (Sequence recognition: No. 306) Con4-26 (C) No inhibition M-Fc-GGGGGAQ- RPEDMCSQWDPWTWHLQGYC-LE (SEQ ID NO: 307) Con4-7 (C) No inhibition M-Fc-GGGGGAQ- LWQLAVCQWDPOTCDHMGAL-LE (Sequence recognition: No. 308) Con4-12 (C) no inhibition M-Fc-GGGGGAQ-TOLVSLCEWDPWTCRLLDGW-LE (sequence recognition · No. 309) Con4-13 (C) no inhibition M-Fc-GGGGGAQ- MGGAGRCEWDPWTCQLLQGW-LE (sequence identification: 310) Con4-14 (C) No inhibition M-Fc-GGGGGAQ-MFLPNECQWDPWTCSNLPEA-LE (Sequence recognition: No. 311) Con4-2 (C) No inhibition M-Fc-GGGGGAQ-FGWSHGCEWDPWTCRLLQGW-LE (Sequence recognition: No. 312) Con4-6 (C) No inhibition M-Fc-GGGGGAQ-WPQTEGCQWDPWTCRLLHGW-LE (SEQ ID NO: 313) Con4-24 (C) No inhibition MF c-GGGGGAQ- PDTRQGCQWDPWTCRLYGMW-LE (Sequence recognition: No. 314) ACl -Con4 (C) No inhibition M-Fc-GGGGGAQ- . TWPODKCEWDPWTCRLLQGW-LE (Sequence recognition别: No. 315) AC3-Con4 (C) No inhibition M-Fc-GGGGGAQ-DKILEECEWDPWTCRLLQGW-LE (Sequence recognition: No. 316) AC5-Con4 (C) No inhibition M-Fc-GGGGGAQ-AATQEECEWDPWTCRLLQGW-LE (sequence Identification: No. 317)

-173- 1317281 (169) I INSTRUCTIONS Continuation of L1-derived Affinity-Mature Pbs hAng-2: Tie2 IC5〇(nM) Peptide Sequence (SEQ ID NO:) Ll-7 (N) 0.03 MGAQ- TNFMPMDDLEQRLYEQFILQQG- LEGGGGG-Fc (SEQ ID NO: 318) AC6-L1 (N) 0.03 MGAQ- TNYKPLDELDATLYEHWILQHS LEGGGGG-Fc (SEQ ID NO: 319) Ll-15 (N) 0.04 MGAQ- QKYQPLDELDKTLYDQFMLQQG LEGGGGG-Fc (Sequence recognition: No. 320) Ll-2 (N) 0.04 MGAQ-LNFTPLDELEQTLYEQWTLQQS LEGGGGG-Fc (SEQ ID NO: 321) L1-10(N) 0.05 MGAQ- QKFQPLDELEQTLYEQFMLQQA LEGGGGG-Fc (Sequence recognition: No. 322) Ll-13 (N) 0.05 MGAQ- QEYEPLDELDETLYNQWMFHQR LEGGGGG-Fc (SEQ ID NO: 323) LI-5 (N) 0.05 MGAQ-VKYKPLDELDEILYEQQTFQER LEGGGGG-Fc (SEQ ID NO: 324) L1-C2 (N) 0.05 MGAQ- TKFQPLDELDQTLYEQWTLQQR LEGGGGG-Fc (Sequence recognition: No. 325) L1-C3 (N) 0.06 MGAQ- TNFQPLDELDQTLYEQWTLQQR LEGGGGG-Fc (SEQ ID NO: 326) Ll-11 (N) 0.07 MGAQ- QNFKPMDELEDTLYKQFLFQHS LEGGGGG-Fc (Sequence recognition: No. 327)

• 174- 1317281 _ (170) Description of the Invention Continued Page L1-17(N) 0.08 MGAQ- VKYKPLDELDEWLYHQFTLHHQ LEGGGGG-Fc (Sequence Identification: No. 328) Ll-12 (N) 0.08 MGAQ- YKFTPLDDLEQTLYEQWTLQHV LEGGGGG-Fc (Sequence Identification: 329 No.) Ll-1 (N) 0.08 MGAQ-QNYKPLDELDATLYEHFIFHYT LEGGGGG-Fc (SEQ ID NO: 330) Ll-4 (N) 0.08 MGAQ- VKFKPLDALEQTLYEHWMFQQA LEGGGGG-Fc (Sequence recognition: No. 331) LI-20 (N) 0.09 MGAQ - EDYMPLDALDAQLYEQFILLHG LEGGGGG-Fc (SEQ ID NO: 332) Ll-22 (N) 0.09 MGAQ- YKFNPMDELEQTLYEEFLFQHA LEGGGGG-Fc (SEQ ID NO: 333) L1-14(N) 0.11 MGAQ- SNFMPLDELEQTLYEQFMLQHQ LEGGGGG-Fc (Sequence recognition: 334 No.) L1-16(N) 0.11 MGAQ- QKFQPLDELEETLYKQWTLQQR LEGGGGG-Fc (sequence identification: 335) · L1-18(N) 0.16 MGAQ-QKFMPLDELDEILYEQFMFQQS LEGGGGG-Fc (sequence identification: No. 336) Ll-3 (N) 0.16 MGAQ- TKFNPLDELEQTLYEQWTLQHQ LEGGGGG-Fc (SEQ ID NO: 337) Ll-21 (N) 0.17 MGAQ- HTFQPLDELEETLYYQWLYDQL LEGGGGG-Fc (Sequence recognition: No. 338)

-175- 1317281 (171) I invention description continuation page

Ll-Cl (N) 0.56 MGAQ- QKFKPLDELEQTLYEQWTLQQR LEGGGGG-Fc (SEQ ID NO: 339) Ll-19 (N) 1.26 MGAQ- QTFQPLDDLEEYLYEQWIRRYH LEGGGGG-Fc (Sequence recognition: 34 nickname) Ll-9 (N) 1.62 MGAQ- SKFKPLDELEQTLYEQWTLQHA LEGGGGG-Fc (SEQ ID NO: 341) Coni-derived affinity-mature Pbs hAng-2: Tie2 IC5〇(nM) Peptide sequence (SEQ ID NO:) Coni-4(C) 1.68 M-Fc-GGGGGAQ -SGQLRPCEEIFGCGTQNLAL-LE (Sequence recognition: No. 342) Conl-1 (C) 3.08 M-Fc-GGGGGAQ- AGGMRPYDGMLGWPNYDVQA-LE (Sequence recognition: No. 343) Coni-6(C) 8.60 M-Fc-GGGGGAQ-GODLRPCEDMFGCGTKDWYG-LE (SEQ ID NO: 344) Coni-3 (C) 16.42 M-Fc-GGGGGAQ-APGQRPYDGMLGWPTYQRIV-LE (Sequence recognition: No. 345) Coni-2 (C) No inhibition M-Fc-GGGGGAQ-QTWDDPCMHILGPVTWRRCI-LE (sequence Identification: No. 346) 'Coni-5 (C) No inhibition M-Fc-GGGGGAQ-FGDKRPLECMFGGPIQLCPR-LE (SEQ ID NO: 347) Parental:: Coni (C) 26.00 M-Fc-GGGGGAQ-KRPCEEIFGGCTYQ-LE ( Sequence identification: No. 348) • 176- 1317281 (172) Hair: _ .··· ;···. -s-ϊ· r*,-.. ::- . , ,., ν. v 12-9 Derived Affinity-Mature Pbs hAng-2: Tie2 ICs〇(nM) Peptide Sequence (SEQ ID NO:) 12-9-3 (Ό 0.81 M-Fc-GGGGGAQ -LOEWCEGVEDPFTFGCEKQR-LE (Sequence recognition: No. 349) 12-9-7 ro 0.93 M-Fc-GGGGGAQ-MLDYCEGMDDPFTFGCDKQM-LE (Sequence recognition: No. 350) 12-9-6 (C) 0.95 M-Fc-GGGGGAQ-HOEYCEGMEDPFTFGCEYQG -LE (Sequence recognition: No. 351) 12-9-C2 (C) 1.41 M-Fc-GGGGGAQ-LODYCEGVEDPFTFGCENQR-LE (Sequence recognition: No. 352) 12-9-5 (C) 1.56 M-Fc-GGGGGAQ-LLDYCEGVQDPFTFGCENLD -LE (Sequence recognition: No. 353) 12-9-1 (C) 1.84 M-Fc-GGGGGAQ-GFEYCDGMEDPFTFGCDKQT-LE (Sequence recognition: No. 354) 12-9-4 (C) 2.05 M-Fc-GGGGGAQ-AODYCEGMEDPFTFGCEMQK -LE (Sequence recognition: No. 355) 12-9-C1 (C) 2.68 M-Fc-GGGGGAQ-LQDYCEGVEDPFTFGCEKQR-LE (Sequence recognition: No. 356) 12-9-2 (C) 8.42 M-Fc-GGGGGAQ-KLEYCDGMEDPFTQGCDNQS -LE (sequence recognition: No. 357) Parent: 12-9 (C) 15.00 M-Fc-GGGGGAQ- . FDYCEGVEDPFTFGCDNH-LE (sequence identification: No. 358)

Example 9 A sample of six anti-Ang2 peptibodies was tested for binding to huAng2 (R&D Systems, BNO12103A) on BIAcore. Protein G was immobilized on a CM5 wafer according to standard amine-conjugated grass (BIAcore Inc.), and then the peptide was injected onto the surface of protein G for capture (RL about 100 Ru). To test the binding between hAng2 and the capture peptide, 0.3 nM to 40 nM of huAng2 177-1317281 (173) was injected onto the surface of the capture peptide body and analyzed using a ΒΙΑ evaluation 3.0 (BIAcore Inc.) binding sensor Figure. Table 8 summarizes the results of this experiment. Table 8 Peptide lot number KD (M) ka (1/Ms) kd (1/s) Con4-44 (C) 011702 2.1E-10 2.9E+05 5.9E-05 Ll-7 (N) 022102 2.4E- 10 3.7E+05 8.7E-05 Ll-10 (N) 021302 7.7E-10 1.5E+05 1.1E-04 Ll-21 (N) 021802 2.4E-10 5.6E+05 1.4E-04 Con4 (C 33456-77 3.8E-10 5.3E+05 2.0E-04 2xCon4 (C) IK 092501 3.4E-10 4.8E+05 1.6E-04 Example 10

Neutralization ELISA Diluted human, mouse, canine and rat Ang-2 ' and human and mouse Ang-ΐ conditioned medium with DMEM/50 μg/ml BSA as follows: hAng_2-i:64 dilution; mAng-2-l:64 Dilution; rat Ang_2_ not diluted; canine 叩_2_1:32 dilution; hAng-ll: 4 dilution; and dilution. By virtue of its 50% achievable maximum binding (ie, the plateau phase), with 1 nM hTie2-Fc (provided by the Tie-2-Fc molecule, where the Tie-2 moiety contains only soluble molecules outside the molecule) Part; R&D Systems, Catalog No. 313-TI) Combining Ability' to determine the degree of dilution of each of these conditioning media. The microtiter plate was coated with 100 microliters of diluted conditioning medium. Regarding the Ang-2 Neutralization ELISA, containing approximately 1% BSA and approximately 1 nM Tie-2 (provided as a Tie-2-Fc molecule, wherein the Tie-2 moiety contains only soluble extracellular portions of the molecule; R& Candidate anti-Ang-2 peptibodies were titrated by 4-fold dilution of PBS solution from D Systems, catalog No. 3 13-TI, from 62.5 nM to 0.015 pM. 1317281 ι_ι (174) | The invention was ironed to Ang-2 Neutralization ELISA to contain approximately 1% BSA and approximately 1 nM Tie-2 (provided as a Tie-2-Fc molecule, wherein the Tie-2 moiety contains only this molecule The soluble extracellular portion; R&D Systems, catalogue No. 3 13-TI) was diluted 4-fold in PBS solution from 1000 nM to 0.2 pM to titrate candidate anti-Ang-2 peptibodies. After adding approximately 100 microliters of peptibody/Tie-2 solution to each well, the plate was incubated overnight at room temperature and then washed 5 times with PBS containing approximately 1% Tween-20. After rinsing, add approximately 100 microliters of anti-Tie-2 antibody per well (卩1^1>111丨11§611111 (:., catalog #55703 9) to approximately 1 μg per ml of final concentration _ degrees, The plate was incubated for approximately 1 hour at room temperature. Next, approximately 100 microliters of goat anti-mouse-IgG-HRP per well was added at a dilution of 1:10,000 in PBS containing approximately 1% BSA (Pierce Chemical) Co., Catalog #31432). Incubate the plate for approximately 1 hour at room temperature, then rinse 5 times with PBS containing approximately 0.1% Tween-20. Then add approximately 100 μl of TMB solution per well (SIGMA, catalogue #T8665), and allow blue to appear. Then absorb the absorbance at 370 nm of the spectrophotometer. The results are shown in Table 9 below. Table 9 Peptide-regulated angiopoietin: Tie2 interactive evaluation Neutralization hAng-2 mAng-2 rAng-2 cAng-2 hAng-1 mAng-2 Peptide IC5〇(nM) IC5〇(nM) IC5〇(nM) IC5〇(nM) IC5〇(nM) IC5〇 (nM) 2xCon4 (C) 0.026 0.035 0.024 0.047 3.0 3.2 Con4 (C) 0.197 0.298 0.236 0.540 200 300 C〇n4-44 (C) 0.08 0.16 0.22 43 «·_ Con4-40 (C) 0.20 0.27 0. 35 > 1000 .... Ll-7 (N) 0.046 0.063 0.035 0.108 > 1000 > 1000 Ll-21 (N) 0.179 0.249 0.204 0.608 > 1000 > 1000 L1-10(N) 0.06 0.06 0.06 — > 1000 — -179- 1317281 (i75) [usa Example 11 PK Study _ Study Shu tf· Male CD-I mice weighing 20-30 grams were randomly assigned to each peptoid treatment group (2xC〇n4-C, Within L1-7-N and L1-21-N), animals received a single IV mass of 50 μg peptibodies (n=38/group) or single administration (n=34/group) for IV and SC administrations respectively Injections were made via the tail vein and under the skin of the shoulder. The predetermined dose measured for each anti-Ang2 peptoid concentration was measured and divided into 5 (: and 1, 4, 4, 8, after administration of the Lu IV group). Blood samples were collected at 16, 24, 48, 72, 96, 120, 144 168, 216, 264, 312 and 336 hours. For group IV, additional samples were collected at 5 and 30 minutes after administration. Two animals were bled at each time point and sacrificed after sampling. From the heart puncture, blood (approximately 0.50 ml) was collected into a polyphosphate such as (10) (6) (iv) 8 serum separation tubes. Place the sample on ice for approximately 20 minutes or until clot formation occurs. The serum was separated from the blood sample by centrifugation at 2-8 C for approximately 1 minute and stored at approximately -7 Torr until the assay was performed. The sample β was coated with the recombinant mouse barley 蛋白质2 protein coated with NUNC fluoroMaxisorp using a Lu, calibrated, time-resolved fluorescence (TRF) assay with a lower limit of quantitation (LL〇Q) of 100 ng/ml. Titration plate. The plate is then blocked with a protein solution to reduce non-specific binding, and standards, quality control groups, and unknown samples are prepared in a 1%% mouse serum assay buffer solution and pipetted to the well of a microtiter plate. in. The peptibodies are specifically combined with the already fixed Ang 2" to rinse off any unbound receptors (Kirkegaard & -180 - 1317281 Invented Tablets (176)

After Perry Laboratories Inc.), biotinylated goat anti-human IgG (H+L) monoclonal antibody (Jackson ImmunoResearch Laboratories Inc.) was added to the wells. After the rinsing step of removing any unbound biotinylated monoclonal antibody, sputum-labeled streptavidin was added to the wells. After rinsing off unbound streptavidin, the acidic solution is pipetted into each well to release the bound streptavidin. A fluorescent signal is generated and read in the Wallaces Fluorescent Reader. The assay for anti-Ang-2 peptibody was analyzed in mouse serum: the circumference was 0.078-5 μg/ml. Drug Dynamics Analysis The integrated mean concentration-time data for each group was subjected to a compartment-free analysis using WinNonlin Professional (version 3.3, Pharsight Corp., Mountain View, CA). For the face sampling time used in the PK analysis, samples were collected within 10% of the face time. All concentration values below LLOQ were set to zero prior to PK analysis. Estimate the following PK parameters:

• Calculate the endpoint half-life at t1/2=In(2)/Kel, where kel is the linear regression of the log-linear variability period through the endpoint to estimate the first-order endpoint rate constant. • Use the linear/logarithmic trapezoid method to estimate the area under the serum concentration-time curve (AUC(〇.last)) from time 0 to the end, the last quantifiable concentration time (Clast). • Evaluate the area under the curve from time 0 to infinity (AUC^oo) by the sum of the corresponding AUC (〇.last) and predicted Clast/ke^): Predicted Clast AUC (〇.〇〇) )=AUC (0-Ust)+ - kel -181 - 1317281 Description of the invention 'Continued (177) • Calculate the absolute bioavailability (F) after SC administration as follows: AUC(〇-〇〇)sc F= + - x 100 AUC (〇.〇〇) iv The results are presented in Figure 2. Example 12

On day 0 of the study, female nude mice were injected subcutaneously with lxlO7 A431 cells. On day 3, the Ang-2 peptide 2xCon4-C was administered subcutaneously at a dose of 200 μg/mouse/day. Tumor volume and body weight were recorded at regular intervals as shown in the figure: Significant differences in tumor growth were observed between the vehicle control group and the control peptibodies in the Ang-2 peptide-treated group (p<0.0001 for each control group, using repeated measures ANOVA, and Scheffe's post hoc assay) ). Treatment with this peptibody did not have a significant effect on body weight. The result is stated in Figure 3. Example 13 Growth curve of A431 in vitro

A431 cells were seeded on 96-well tissue culture plates in 2000 microliters per well in 200 microliters of DMEM supplemented with 10% fetal bovine serum (FBS). After sowing, the medium was evacuated for 16 hours. The following materials were then added back to the wells and established in triplicate: 100 microliters of DMEM per well, 10% FBS, 1 mg/ml negative control peptibody 4883, or peptidomimetic TN8-Con4. Repeat the same combination on 5 plates. The medium from one plate was evacuated at 24, 48, 72, 96 and 120 hours after the treatment. Then, 100 μl of 10% trichloroacetic acid (TCA) was added to each well, and the plate was then stored at 4 °C. All plates were collected and the last plate was at least 4 hours in 10% TCA -182-1317281 (178). Drain 10% TCA' and rinse each well 5 times with tap water and then at room temperature with 100 μl of 0.4% sulphur rhodamine B (Sigma S-9012) in 1% acetic acid (Sigma Α-6283) The cells were stained for 1 minute and washed 5 times with 1% ethyl red. The plate is then air dried. The dye was solubilized for 2 hours on a rotary vibrator using 3 Torr of 20 mM unbuffered Tris (pH > 10). The optical density (〇D) was then read at 540 nm at the microtiter plate reader. The result is stated in Figure 4. Example 14

Female nude mice were injected subcutaneously with 2x10 Colo-205 cells plus Matrigel (2:1) on study day 0. On day 3, at a dose of 14 μg/mouse, two weeks after the human skin was again subcutaneous with the Ang-2 skins L1-7-N, L1-21-N, Con4-C and 2xCon4-C. Anti-Ang-2 antibody, Ab536, 47 μg/mouse was administered three times a week as a positive control group. Tumor volume and body weight were recorded at regular intervals.

Significant differences in tumor growth were observed between each of the vehicle control group and the control peptibosome in each Ang-2 peptidial treatment group (p<1 for each control group, using repeated measures ANOVA, With Scheffe's post hoc accreditation). Treatment with these peptibodies had no significant effect on body weight (results not shown). The result is stated in Figure 5. Example 15 Female nude mice were injected subcutaneously with 2 χ 10δ Colo-205 cells plus Matrigel (2:1) at study day 0. On day 3, Ang-2 peptidomimetic 2xCon4-C was administered subcutaneously twice a week at doses of 14, 2.8 and 0.56 μg/mouse. Depending on the regulatory interval shown, § Tumor volume and body weight have been recorded. Between the two higher doses of the Ang_2 peptoid treatment group, between the vehicle control group and the control peptide, tumor-183-1317281 (179) was observed. Continued _ ., . , .1 . . Significant differences in AJv growth (p=0.003 for intermediate dose, and p<0001 for high dose, using repeated measures ANOVA, with Scheffe's post hoc assay). Treatment with these peptides had no significant effect on body weight. The dashed line represents the total η value of the group, which decreased from 10 to 9 mice, due to a mouse that died unexplained. The result is stated in Figure 6. Example 16

Tumors of anti-Ang-2 peptidoglycan on Colo-205 xenograft On day 0 of the study, 2 x 106 Colo-205 cells plus Matrigel (2:1) were subcutaneously injected: female nude mice were injected. On day 3, Ang-2 peptidomimetic 2xCon4-C or control peptibody was administered subcutaneously at a dose of 350 μg/day. Tumors from the group treated with the control peptoids were obtained on day 14 (size-match control group) or day 18 (time-match control group) (as described in Table 5). On day 18, tumors obtained from 2xCon4(C) were recorded at regular intervals as shown, and tumor volumes were recorded. Between the time-matched control group and the 2xCon4-C treatment group,

Significant differences in tumor growth were observed (p=0.0154, with an emphasis on ANOV, and Scheffe's post hoc assay). Treatment with these peptibodies did not have a significant effect on body weight. The tumor prepared for image analysis was cut from the top portion to two and half was suddenly cold; east in OCT (Sakura Finetek USA Inc., Torrance, CA). Frozen-slices were stained immunohistochemically using 2 μg/ml anti-mouse CD31 (catalog #553370, BD PharMingen, San Diego, CA) with DAB as chromogen. Digital photos of tumor sections were taken at 20X objective magnification. Capturing the four "Bound-Points" fields for each tumor, each treatment group has 10 tumors. For the threshold of blood vessels stained with CD3 1 in the image -184 - 1317281

1 Λ . ι·..,. Shoulder 1, :η. * ,: -« ftSfiSAS (180), using MetaMorph (Universal Imaging Corporation, Downington, PA) image analysis system. The area of CD3 1 positive staining was expressed in proportion to the total tumor tissue in each field. The result is stated in Figure 7. Example 17

On day 0 of the study, female nude mice were injected subcutaneously with 2 x 106 Colo-205 cells plus Matrigel (2:1). From the 3rd, 10th or 15th day of the study, Ang-2 peptibo 2xCon4-C was treated with 350 μg/mouse, subcutaneously twice a week, or with an equivalent control peptibody. Record tumor volume and body weight at regular intervals. Significant differences in tumor growth were observed between all Ang-2 peptidial treatment groups versus vehicle control groups (Phase 15 day group, p=0.089, and Days 3 and 10, p<0.0001, heavy use) Overlay measurement ANOVA, with Scheffe's post hoc test). Treatment with these peptibodies did not have a significant effect on body weight. The results are shown in Figure 8 (weight is not shown). Example 18

In the A431 and Colo-205 xenograft mode, antibody Ab53 6, administered at a dose of 47 μg/female nude mice, was administered intraperitoneally three times a week, or in different long-term studies (> 10 weeks of dosing). A multi-drug schedule was given to give a summary of the complete response (CR) rate of the skin 2xCon4(C)'. As used herein, CR' means the result of a measurable tumor remaining after treatment. The result is stated in Figure 9. Example 19 In the C〇1〇_205 tumor model, mixed Pb plaques were injected subcutaneously into female nude mice at 2×10 6 Colo-205 cells plus Matrigel (2:1). On study day 14 'start with a) 350 μg/mouse, -185- 1317281 (181) Invention Description Continue Xi subcutaneous 'twice a week Ang-2 peptibo 2xCon4-C, b) 20 mg/kg three times a week , the cancer in the abdominal cavity is easy, or c) mixed treatment. Tumor volume and body weight were recorded at regular intervals. Significant differences in tumor growth were observed between all treatment groups versus vehicle control groups (p<; 〇〇〇 , 1, using repeated measures ANOVA' and Scheffe's post hoc assay). In addition, there was a significant difference between the mixed treatment group and any single therapeutic agent (p < 〇〇〇〇 1 versus 2xCon4-C ' and p = 0.0122 for gram cancer easily) The dashed line represents the total η value of the group 'from 10 Reduced to 9 mice, due to a mouse die unexplained: Death. Treatment with these peptibodies had no significant effect on body weight. The results are presented in Figure 10a.

b) In the Colo-205 tumor model, mixed pb舆5-FU was injected subcutaneously into female nude mice at 2x106 Colo-205 cells plus Matrigel (2:1) on study day 0. On study day 14, start with a) 350 μg/mouse, subcutaneous, twice a week of Ang-2 peptibody 2xCon4-C, b) 50 mg/kg, 5 times a week, intraperitoneal 5-FU 'or c Both are mixed. Tumor volume and body weight were recorded at the regular intervals shown. Significant differences in tumor growth were observed between all treatment groups versus vehicle control groups (ρ < 0.00 (Η, using repeated measures ANOV, and Scheffe's post hoc assay). In addition, the mixed treatment group and either single treatment There were significant differences in the agents (P=〇.〇375 vs. 2xCon4-C' and p=〇.〇453 vs. 5-FU). In the 5-FU treatment group (18% in the study 20 days) and the mixed treatment group (A temporary decrease in body weight was observed in the 16% of the study on the 2nd day of the study) and then the body weight was completely restored. The results are stated in Figure 1 Ob. -186- 1317281 ...,

(182) IISH Adjuvant Arthritis, male Lewis rats (120-130 g in a 12-hour light/dark cycle environmental control room (temperature 23 ± 2 ° C, relative humidity 50 ± 20%) , Charles River, Wilmington ΜΑ) is housed in two cages covered with filter paper. The caries animal food (formulation 8640; Tek Lab, Madison, WI) is commercially available and can receive filter paper-purified tap water without restriction. The levels of calcium and phosphorus in the diet were 1.2% and 1.0%, respectively.

A single intradermal injection of 0.5 mg of heat-killed Mycobacterium tuberculosis H37Ra (Difco Laboratories, Detroit, MI) in a 5 ml paraffin oil: (Crescent Chemical Co., Hauppauge, NY) ), inducing adjuvant arthritis. On the 9th day, clinical symptoms of arthritis represented by swelling of the hind paw and difficulty in walking occurred. Except for the 2xCon4(C) treatment group (treated from day 1 after immunization), 'on the 9th day after immunization, the daily subcutaneous injection (before the onset of arthritis) was given, and continued until the 18th day. . Adjuvant arthritis of the skin. In clinically, by intermittently measuring the volume of the hind paw, use according to Feige

Et al., Cellular Molec. Life Sci., 57: 1457-1470 (2000) describes the method of water plethysmography to assess the progression of inflammation based on the area under the curve (AUC), using the following formula Trapezoidal rule to calculate the inhibition of paw inflammation: [1-{(Processed AdA)-normal value)/(Untreated AdA_normal value)}]χ100 In addition, during the 9-day treatment regimen Within the day, the overall weight is judged as a supplementary end point, because in the arthritis mode, weight loss has been shown to proceed in parallel with joint inflammation. On the 18th day, sacrifice animals under C02 -187- 1317281 __ face (183)

At the time of post-mortem (18th day after immunization), the loss of bone mineral density (BMD) was examined. Remove the hind paw at the soft hair boundary (near the ankle (after the ankle)), soak in 70% ethanol, and then use a fan beam X-ray densitometer in the horizontal direction (QDR-4500A; Hologic, Waltham, MA) scan. See Feige et al., before. After the scan, place the rectangular box (29x25 mm) centered on the calcaneus to depict the contour of the location to be analyzed and calculate the bone area and bone minerals using a patented algorithm (Hologic software). Intra-substance content and bone mineral density. All results are expressed as mean soil standard deviation. A significant difference between the groups is described using a P value of 0.05. Kruskal-Wallis ANOVA and Mann-Whitney U. assays were performed on clinical data (continuous variables) using commercially available statistical software (StatSoft version 3.0; StatSoft, Tulsa, OK). The results are presented in Figures la, 1 lb and 1 lc, respectively. Example 21 Corneal Angiogenesis Pattern

Effect of VEGF-guided angiogenesis in rats Ang-2 peptibody Con4 (C) was evaluated in the angiogenic cornea model of rats. Angiogenesis was induced by implantation of a VEGF- (or BSA control) soaked nylon disc into the corneal stroma (n=8/group). The peptibody TN8Con4-C was administered 1.0 or 0.1 mg/rat/day by subcutaneous injection for 7 days. The other two groups of animals were treated with the same dose of the negative control peptibody 4883. All groups were pre-treated with a single loading dose of 3.0 or 0.3 mg, which was 3 times the maintenance dose of 1.0 or 0.1 mg (see figure). After 7 days of treatment, from the digital photograph of each rat cornea, the two vascular end points were determined: at the mid-point between the circle -188-1317281 (184) __ disc and the corneal sclera junction The number of blood vessels, as well as the area of the blood vessels. Treatment with TN8CON4-C significantly inhibited VEGF-induced angiogenesis (p<0.04) in a dose-dependent manner, whereas treatment with the control group had no significant effect on either endpoint. Based on the body weight of the treated animals, there is no evidence of significant toxicity. The result is illustrated in Figure 12. Example 22 epitope mapping

The human Ang-2 (hAng-2) protein of full length (amino acid 1-495), N-terminal (amino acid 1-254) and C-terminal (amino acid 255-495) was selected into the band. There is a C-terminal 6xHis tag in a CMV-driven mammalian expression vector. Three resulting constructs were transiently expressed in 293T cells, and the control group was loaded. The conditioned medium was then collected from the transfected cells and the degree of expression of Ang-2 in the medium was assessed by anti--6xhis ELIS A and Western blotting.

The binding epitopes of anti-Ang-2 antibodies and peptibodies were determined by ELISA according to the following draft, by their ability to bind to three versions of human hAng-2: coating high-binding with 100 microliters of conditioning medium The 96-well wells were assayed for each well and incubated for 1 hour at 37 °C. The conditioning medium was aspirated and the plate was blocked for 1 hour at room temperature with 200 microliters of 5% BSA in PBS per well. The blocking solution is then aspirated. At 1 μg/ml (in 1% BSA in PBS), 100 μl of antibody, peptibody or Tie2-Fc per well was added and incubated for 1 hour at room temperature. The well was rinsed 4 times with 200 microliters of 0.1% Tween in PBS. Add 100 μl of HRP-conjugated goat anti-human IgG or goat anti-aging to each well • 189- 1317281 (185)

Murine IgG was incubated for 45 minutes at room temperature. The well was then rinsed 4 times with 200 microliters of 0.1% Tween in PBS. Then 100 microliters of TMB substrate was added to each well. The O.D. value was read at 370 nm. The results are set forth in Figures 13a, 13b and 13c, respectively. Example 23 Because of certain sensitivity limitations in the BiaCore assay, the Sepidyne KinExA assay was also used to assess binding affinity.

The binding of 2xCON4-C (Pb5714) to -huAng-2 was tested on KinExA (Sapidyne, Boise, ID). Reacti-Gel 6x beads (Pierce, Rockford, IL) were pre-coated with huAng-2 and blocked with BSA. Samples of 10 pM and 30 pM of 2xCON4-C were incubated with various concentrations (0.3 pM - 3 nM) of huAng-2 for 8 hours at room temperature before running the beads coated with huAng-2. The amount of peptibosome bound to the beads was quantified by Lua's (Cy5) labeled goat anti-human-Fc antibody (Jackson Immuno Research, West Grove, PA). The binding signal is proportional to the concentration of the free peptibosome at equilibrium.

The dissociation equilibrium constant (KD) was obtained from the nonlinear regression of the competitive curve using a two-curve single-position uniform binding mode (KinExTM software). It was then determined that approximately 2 pM of 2xCON4-C binds to HuAng-2. As shown in Figure 14, using the KinExA assay, it was shown that the body 2xCon4 has an affinity of about 2 pM for hAng-2. Example 24 PEGylated at with a 43 1 ABI synthesizer using standard coupling drafts and double coupling, from residue 14 (met) to N-terminal residue l (Cys) (from N_terminal to (: _Terminal number), -190- 1317281 (186) Invention said continuation page! li. 3 Synthesis of L1-7 peptide. L1-7 peptide 舆-methyl-poly(ethylene glycol maleimide) Yoke effect; molecular weight: 5KDa; called "mPEG5K-(Ll-7 peptide

Using 13.5 mg of methoxy-poly(ethylene glycol)-maleimide (molecular weight = 5 KDa; Shearwater Corp.) (0.27 ml of 50.0 mg/ml solution in buffer solution 1), treated at 400 μm A solution of 0.8 mg of L1-7 peptide in buffer solution 1 (20 mM phosphate, 5 mM EDTA, pH 6.5). The reaction mixture was incubated at 4 ° C overnight, then diluted with 1.6 ml of buffer solution A (20 mM: Tris hydrochloride, pH 7.2) and in a Slide-A-Lyzer cassette (3500 MWCO, Pierce), Dialysis of the same buffer solution. The dialyzed reaction mixture was purified by ion exchange chromatography on a 1.0 ml HiTrap Q agarose HP column (Amersham Biosciences Corp.). The product peak eluted in two 1.0 ml fractions over a gradient of more than 40 column volumes via a gradient from 100% buffer solution A to 100% buffer solution B (buffer solution A + 0.5 M NaCl). The combined product fractions were concentrated to 250 microliters using a Microsep 1K centrifuge (Pall Life Sciences) containing 0.23 mg protein/ml. Conjugation of L1-7 peptide 舆1.11-bis-maleimidoiminotetraethylene glycol; called "? Ugly 04 (!^1-7 peptide>)," with 0.0175 mg of 1,11-bis-cis-butanediamine-imidotetraethylene glycol (Pierce) (0.375 ml of 0.1 in buffer solution 1) a milligram/ml solution), a solution of 1.0 mg of L1-7 peptide in 500 μl of buffer solution 1 (20 mM phosphate, 5 mM EDTA, pH 6.5) was treated. The reaction mixture was incubated at 4 ° C for 3.33 hours. It was then dialyzed against buffer solution A (20 mM Tris hydrochloride, pH 7.2) in a Slide-A-Lyzer cassette (3500 MWCO, Pierce) by ion exchange -191 - 1317281 (187) chromatography. The dialyzed reaction mixture was purified on a 1.0 ml HiTrap Q ί-lipose HP column (Amersham Biosciences Corp.) via a gradient from 100% buffer solution A to 100% buffer solution B (buffer solution A + 0.5 M NaCl) The product peak of the dimer eluted in three 1.0 ml fractions over 40 column volumes. The mixed product fractions were concentrated to 550 microliters using a Microsep 1K centrifuge (Pall Life Sciences). Contains 0.12 mg protein/ml.

Conjugation of L1-7 peptide with poly(ethylene glycol V bis-maleimide; molecular weight: 3.4KDa; called "PEG3.4K (X1-7 peptide V'

1.125 mg of poly(ethylene glycol)-bis-cis-butane quinone imine (molecular weight = 3.4 KDa, Shearwater Corp.) (0.563 ml of a 2.0 mg/ml solution in buffer solution 1), treated at 1.5 ml A solution of 3.0 mg of L1-7 peptide in buffer solution 1 (20 mM phosphate, 5 mM EDTA, pH 6.5). The reaction mixture was incubated at 4 ° C overnight, and then dialyzed against buffer solution A (20 mM Tris hydrochloride, pH 7.2) in a Slide-A-Lyzer cassette (3500 MWCO, Pierce). The dialyzed reaction mixture was purified by ion exchange chromatography on a 5.0 ml HiTrap Q Sepharose HP column (Amersham Biosciences Corp.). Product peaks were eluted in three 3.0 ml liters of the fraction over a volume of more than 40 column volumes via a gradient from 100% buffer solution A to 100% buffer solution B (buffer solution A + 0.5 M NaCl). The combined product fractions were concentrated to 850 μL using two Microsep 1K centrifuges (Pall Life Sciences) containing 0.24 mg protein/ml. The results of MALDI-TOF mass spectrometry are as follows: -192- 1317281 (188) • ..·· -;' ,-· 'V;·. V ' - invention says _ sample number identity MS experimental value MS observation value 1 L1-7 (unpegylated peptide) 3,545 3,538.7 2 mPEG5K-(Ll-7 peptide) 8.500 8,851 3 PE04 (Ll-7 peptide) 2 7,443 7,446.29 4 PEG3.4K (Ll-7 peptide) 2 10,550 10,552 6,882.61 3,550.13

It should be understood that the subscripts "2" of PEG3.4K (Ll-7 peptide) and PE04 (Ll-7 form) represent two peptides per polymer chain, located at both ends of the polymer. 1 £5 〇 judgment

The IC50 of inhibition of hAng-2:hTie2-Fc interaction was determined by the neutralization of the L1-7 and PEGylated peptides as described in Example 2 and ELISA. For neutralization ELISA, microtiter plates that have bound human Ang-2 polypeptide were prepared as described for the affinity ELISA in Example 2. Titration of candidate anti-Ang-2 PEGylated L1-7 and free peptides from 1〇〇〇nM to 0.2 pM to contain approximately 1% BSA and approximately 1 nM Tie-2 (with Tie-2-Fc molecule) The form is provided in which the Tie-2 portion contains only the soluble extracellular external fraction of the molecule; R&D Systems, catalog No. 3 13-TI) in a 4-fold dilution of PBS solution. After adding approximately 100 microliters of antibody/Tie-2 solution to each well, the plate was incubated overnight at room temperature and then washed 5 times with PBS containing approximately 0.1% Tween-20. After rinsing, add approximately 1 μL/well of anti-Tie-2 antibody (Pharmingen Inc., catalog #557039) to a final concentration of approximately 1 μg/ml and incubate the plate at room temperature approximately 1 hour. Next, dilute with 1:1 〇, 〇〇〇 containing approximately 1% BSA in PBS, add approximately 100 μl/well of -193-1317281 (189) of goat anti-mouse-IgG-HPR (Pierce Chemical Co ., catalog #31432). The plate was incubated for about 1 hour at room temperature and then washed 5 times with PBS containing about 0.1% Tween-20. Approximately 100 microliters/well of TMB substrate (described above) was then added and color rendering was allowed. The absorbance was then read at 370 nm in a spectrophotometer. The L1-7 peptide (C-GGGGG-AQ-TNFMPMDDLEQRLYEQFILQQG-LE) (SEQ ID NO: 359) includes: N-terminal cysteine coupled to PEG; and 5Gly linker. Both the AQ and LE side sequences are present in the original phage pure line and the peptide. hAng-2: Tie2 inhibition IC50 results are as follows: peptide IC5 〇 (nM) L1-7 peptide 0.49 mPEG5K- (Ll-7 peptide) 11.7 PE04 (Ll-7 peptide) 2 0.064 PEG3.4K (Ll-7 peptide) 2 0.058 -194- Description of Inventions Continuation Page η ·· : ";··. · :--·:,·····.: ·- -V? 1317281 (190) Sequence List <110> AMGEN INC. < 120> a specific binding agent for human angiopoietin-2

<130> A-810A <140> has not been assigned <141> 2002-10-11

<150>US (not yet assigned) <151> 2002-09-27 <150> US 60/328,624 <151> 2001-10-11 <160>359

<170> PatentIN Version 3.1 <210> 1 <211> 14 <212> Protein <213> Artificial Sequence <220><223> Ang-2 Binding Peptide-195- (191) 1317281 <;400> 1

Lys Arg Pro Cys Glu Glu Met Trp Gly Gly Cys Asn Tyr Asp 15 10 <210> 2 <211> 14 ' <212> Protein <213> Artificial Sequence <220>

<223> Ang-2 binding peptide <400> 2

His Gin lie Cys Lys Trp Asp Pro Trp Thr Cys Lys His Trp 1 5 10 <210> 3 <211> 14 <212> Protein <213> Artificial Sequence <220>

<223> Ang-2 binding polypeptide <400> 3

Lys Arg Pro Cys Glu Glu lie Phe Gly Gly Cys Thr Tyr Gin 15 10 <210> 4 <211> 14 <212> Protein <213> Artificial sequence -196· 1317281 (192) Invention says contiguous page % <220><223> Ang-2 binding polypeptide <400> 4

Gin Glu Glu Cys Glu Trp Asp Pro Trp Thr Cys Glu His Met 1 5 10 <210> 5 <211> 18 <212> Protein <213> Artificial Sequence <220><223> Ang-2 Binding polypeptide <400> 5

Cys Asp 15

Phe Asp Tyr Cys Glu Gly Val Glu Asp Pro Phe Thr Phe Gly 15 10

Asn His <210> 6 <211> 20 <212> Protein <213> Artificial sequence <220><223> Ang-2 combined polypeptide <400>

Gin Phe 15

Lys Phe Asn Pro Leu Asp Glu Leu Glu Glu Thr Leu Tyr Glu 15 10

Thr Phe Gin Gin 20 197- 1317281 (193) Description of the invention Net page 3 ................ -'r.ii..'. . . . 1 <210> 7 <211> 14 <212> Protein <213> Artificial sequence <220><223> Ang-2 binding peptide <400>

Gin Tyr Gly Cys Asp Gly Phe Leu Tyr Gly Cys Met work le Asn 15 10 <210> 8 <211> 67

<212> DNA <213> Artificial sequence <220><223>oligonucleotide<400> 8 acaaacaaac atatgggtgc acagaaagcg gccgcaaaaa aactcgaggg tggaggcggt 60

Ggggaca 67 <210> 9 <211> 20 <212> DNA <213> Artificial sequence <220><223> Oligonucleotide-198- 1317281 (194) <400> 9 ggtcattact ggaccggatc <210> 10 <211> 25 <212> . DNA ' < 213 > artificial sequence <220><223> Oligonucleotide <400> 10 cgtacaggtt tacgcaagaa aatgg <210> 11 <;211> 66 <212> DNA <213> Artificial Sequence Invention Description Continued Page j - 20 25

<220><223> Oligonucleotide <400> 11 tttgttggat ccattactcg agtttttttg cggccgcttt ctgtgcacca ccacctccac ctttac <210> 12 <211> 32 <212> Protein <213> Artificial sequence <220><223> Peptide 60 66 capable of binding to Ang-2

-199- (195) 1317281 ΐϋ _ <220><22i> misc - feature <222> (32)..(32) <223> Xaa = Fc <400> 12

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

Tyr Glu Gin Phe Thr Phe Gin Gin Leu Glu Gly Gly Gly Gly Gly Xaa 20 25 30 <210> 13 <211> 29. <212> Protein <213> Artificial Sequence <220><223> Peptide capable of binding to Ang-2 <220>

<22i> misc_ feature <222> (29) .. (29) <223> Xaa = Fc <400> 13

Met Lys Phe Asn Pro Leu Asp Glu Leu Glu Glu Thr Leu Tyr Glu Gin 15 10 15

Phe Thr Phe Gin Gin Leu Glu Gly Gly Gly Gly Gly Xaa 20 25 <210> 14 200- 1317281 (196) Inventive Note Continued] <211> 51 <212> Protein <213> Artificial Sequence<220><223> Peptide capable of binding to Ang-2 <220><22i> misc_ feature

<222> (51)..(51) <223> Xs.h = F* c <400> 14 Met Lys Phe Asn Pro Leu Asp Glu Leu Glu Glu Thr Leu Tyr' Glu Gin 1 5 10 15 Phe Thr Phe Gin Gin Gly Ser Gly Ser Ala Thr Gly Gly Ser Gly Ser 20 25 30 Ala Ser Ser Gly Ser Gly Ser Ala Thr His Leu Glu Gly Gly Gly 35 40 45

Gly Gly Xaa 50 <210> 15 <211> 60 <212> Protein <213> Artificial sequence <220><223> Peptide capable of binding to Ang-2 <220> 1317281 (197) Description of the invention: Poverty reduction: <22i> misc_ feature <222> (60). (60) <223> Xaa = Fc <400> 15 '

Met Gly Ala Gin Lys Phe Asn Pro Leu Asp Glu Leu Glu Glu Thr Leu 1 5 l〇 15

Tyr Glu Gin Ghe Ghe Gly Gly Gly Gly Gly Gly 20 25 30

Lys Phe Asn Pro Leu Asp Glu Leu Glu Glu Thr Leu Tyr Glu Gin Phe 35 40 45

Thr Phe Gin Gin Leu Glu Gly Gly Gly Gly Gly Xaa 50 55 60 <210> 16 <211> 56 <212> Protein <213> Artificial Sequence <220><223><220> Peptide bound to Ang-2 <221> misc_ feature <222> (56)..(56) <223> Xaa = Fc <400>

Met Lys Phe Asn Pro Leu Asp Glu Leu Glu Glu Thr Leu Tyr Glu Gin 15 10 15 •202· (198)1317281 Description of the Invention Continued: ··· . ·.. -乂:

Phe Thr Phe Gin Gin Gly Gly Gly Gly Gly Gly Gly Lys Phe Asn Pro 20 25 30

Leu Asp Glu Leu Glu Glu Thr Leu Tyr Glu Gin Phe Thr Phe Gin Gin 35 40 45

Leu Glu Gly Gly Gly Gly Gly Xaa 50 55 <210> 17 <211><212><213> 26 Protein artificial sequence <220><223> Peptide capable of binding to Ang-2 <;220><221> misc_ feature <222> (26)..(26) <223> Xaa * Fc <400> 17

Met Gly Ala Gin Gin Glu Glu Cys Glu Trp Asp Pro Trp Thr Cys Glu 15 10 15

His Met Leu Glu Gly Gly Gly Gly Gly Xaa 20 25 <210> 18 <211> 45 <212> Protein <213> Artificial Sequence 203 - 1317281 _ (199) I Invention Description Continued <220><223> Peptide capable of binding to Ang-2 <220><22i> misc_ feature <222> (45)..(45) <223> Xaa «Fc <400>

Met Gin Glu Glu Cys Glu Trp Asp Pro Trp Thr Cys Glu His Met Gly 15 10 15

Ser Gly Ser Ala Thr Gly Gly Ser Gly Ser Thr Ala Ser Ser Gly Ser 20 25 30

Gly Ser Ala Thr His Leu Glu Gly Gly Gly Gly Gly Xaa 35 40 45 <210> 19 <211> 62 <212> Protein <213> Artificial Sequence

<220><223> Peptide capable of binding to Ang-2 <220><22i> misc_feature <222> (62)* (62) <223> Xaa = Fc <400>; 19

Met Gly Ala Gin Gin Glu Glu Cys Glu Trp Asp Pro Trp Thr Cys Glu 15 10 15 -204- (200) 1317281 Description of the Invention Continued ^

His Met Gly Ser Gly Ser Ala Thr Gly Gly Ser Gly Ser Thr Ala Ser 20 25 30

Ser Gly Ser Gly Ser Ala Thr His Gin Glu Glu Cys Glu Trp Asp Pro 35 -40 45

Trp Thr Cys Glu His Met Leu Glu Gly Gly Gly Gly Gly Xaa 50 55 60 <210> 20 <211> 31

<212> Protein <213> Artificial sequence <220><223> Peptide capable of binding to Ang-2 <220><221> misc-feature <222> (2).. ( 2) <223> Xaa = Fc <400> 20

Met Xaa Gly Gly Gly Gly Gly Ala Gin Lys Phe Asn Pro Leu Asp Glu 15 10 15

Leu Glu Glu Thr Leu Tyr Glu Gin Phe Thr Phe Gin Gin Leu Glu 20 25 30 <210> 21 <211> 53 <212> Protein <213> Artificial Sequence - 205 - 1317281 (201) Gong <220><223> Peptide capable of binding to Ang-2 <220><221> misc_ feature <222> (2)..(2) <223> Xaa = Fc <400&gt ; twenty one

Met Xaa Gly Gly Gly Gly Gly Ala Gin Gly Ser Gly Ser 15 10

Gly Ser Gly Ser Thr Ala Ser Ser Gly Ser Gly Ser Ala 20 25

Phe Asn Pro Leu Asp Glu Leu Glu Glu Thr Leu Tyr Glu 35 40 45

Phe Gin Gin Leu Glu 50 <210> 22 <211> 59 <212> Protein <213> Artificial sequence <220><223> Peptide capable of binding to Ang-2 <220>;22i>misc_features<222> (2)..(2) <223> Xaa = Fc

Ala Thr Gly 15

Thr His Lys 30

Gin Phe Thr •206- 1317281 (202) Skillfully said continuation page ^ <400> 22

Met Xaa Gly Gly Gly Gly Gly Ala Gin Lys Phe Asn Pro Leu Asp Glu 15 10 15

Leu Glu Glu Gly Gly Gly Gly 20 25 30 Leu Glu Glu Gin Ghe Gly Gly 20 25 30

Gly Gly Gly Gly Gly Lys Phe Asn Pro Leu Asp Glu Leu Glu Glu Thr 35 40 45

Leu Tyr Glu Gin Phe Thr Phe Gin Gin Leu Glu 50 55 <210> 23 <211> 25 <212> Protein <213> Artificial sequence <220><223> capable of binding to Ang-2 Peptide <220><221>misc-feature<222> (2).. (2) <223> Xaa = Fc <400> 23

Met Xaa Gly Gly Gly Gly Gly Ala Gin Gin Glu Glu Cys Glu Trp Asp 15 10 15

Pro Trp Thr Cys Glu His Met Leu Glu 20 25 <210> 24 207- 1317281 __ (203) Description of the Invention Continued: <211> 47 <212> Protein <213> Artificial Sequence <220><;223> Peptide capable of binding to Ang-2 <220〉

<22i> misc_ feature <222> (2)..(2) <223> X33. — F c <400> 24

Met Xaa Gly Gly Gly Gly Gly Ala Gin Gly Ser Gly Ser Ala Thr Gly 15 10 15

Gly Ser Gly Ser Thr Ala Ser Ser Gly Ser Gly Ser Ala Thr His Gin 20 25 30

Glu Glu Cys Glu Trp Asp Pro Trp Thr Cys Glu His Met Leu Glu 35 40 45

<210> 25 <211> 61 <212> Protein <213> Artificial sequence <220><223><220> Peptide capable of binding to Ang-2 <221><222> (2) · · (2) -208- 1317281 (204) Description of the invention page) <223> Xaa = Fc <400> 25

Met Xaa Gly Gly Gly Gly Gly Ala Gin Gin Glu Glu Cys Glu Trp Asp 1 , 5 10 15

Pro Trp Thr Cys Glu His Met Gly Ser Gly Ser Ala Thr Gly Gly Ser 20 25 30

Gly Ser Thr Ala Ser Ser Gly Ser Gly Ser Ala Thr His Gin Glu Glu 35 40 45

Cys Glu Trp Asp Pro Trp Thr Cys Glu His Met Leu Glu 50 55 60 <210> 26 <211> 75 <212> Protein <213> Artificial Sequence <220><223> 2-bound peptide body <220><221> misc_ feature <222> (75)..(75) <223> Xsd. = Fc <400>

Met Gly Ala Gin Glu Glu Cys Glu Trp Asp Pro Trp Thr Cys Glu His 15 10 15

Met Gly Gly Gly Gly Gly Gly Gly Gly Gly Lys Phe Asn Pro Leu Asp Glu 20 25 30 -209 (205) (205) 13172281 Invention: Alum page 3

Leu Glu Glu Thr Leu Tyr Glu Gin Phe Thr Phe Gin Gin Gly Ser Gly 35 40 45

Ser Ala Thr Gly Gly Ser Gly Ser Thr Ala Ser Ser Gly Ser Gly Ser 50 55 60

Ala Thr His Leu Glu Gly Gly Gly Gly Gly Xaa 65 70 75 <210> 27 <211> 72 <212>Protein<213> Artificial sequence <220><223> can be combined with Ang-2 Peptide <220><221> misc_feature <222> (2)..(2) <223> Kaa = Fc <400>

Met Xaa Gly Gly Gly Gly Gly Ala Gin Gly Ser Gly Ser Ala Thr Gly 15 10 15

Gly Ser Gly Ser Thr Ala Ser Ser Gly Ser Gly Ser Ala Thr His Lys 20 25 30

Phe Asn Pro Leu Asp Glu Leu Glu Glu Thr Leu Tyr Glu Gin Phe Thr 35 40 45

Phe Gin Gin Gly Gly Gly Gly Gly Gin Glu Glu Cys Glu Trp Asp Pro 50 55 60

Trp Thr Cys Glu His Met Leu Glu 65 70 -210- (206) 13172281 Summary of the Invention Page: <210> 28 <211> 30 Protein <213> Artificial Sequence <220><223> Ang-2 bound peptide body <220>

<22i> misc - feature <222> (30), (30) <223> Xaa = Fc <400> 28

Met Gly Ala Gin Phe Asp Tyr Cys Glu Gly Val Glu Asp Pro Phe Thr 15 10 15

Phe Gly Cys Asp Asn His Leu Glu Gly Gly Gly Gly Gly Xaa 20 25 30 <210> 29

<211> 26 <212> Protein <213> Artificial sequence <220><223> Peptide capable of binding to Ang-2 <220><22i> misc_ feature <222> )..(26) -211 - 1317281 (207) Description of the Invention Continued Page 4 <223> Xaa = Fc <400> 29

Met Gly Ala Gin Gin Tyr Gly Cys Asp Gly 1 . 5 l〇lie Asn Leu Glu Gly Gly Gly Gly Gly Xaa 20 25 <210> 30 <211> 26 <212> Protein <213> Artificial Sequence <;220><223> Peptide capable of binding to Ang-2 <220><22i> misc - feature <222> (26) (26) <223> Xaa == Fc

Phe Leu Tyr

Gly Cys Met 15

<400> 30 Met Gly Ala Gin Lys

Arq Pro Cys Glu Glu 10

Met Trp Gly Gly Cys Asn 15

Tyr Asp Leu Glu Gly Gly Gly Gly Gly Xaa 20 25 <210> 31 <211> 26 <212> Protein <n3> Artificial Sequence 212-1317281 Description of the Invention Continued Page (208) <220> 223> Peptide capable of binding to Ang-2 <220><22i> misc_ feature <222> (26)..(26) <223> Xaa = Fc <400>

Met Gly Ala Gin His Gin lie Cys Lys Trp Asp Pro Trp Thr Cys Lys 15 10 15

His Trp Leu Glu Gly Gly Gly Gly Gly Xaa 20 25 <210> 32 <211> 26 <212>Protein<213>. Artificial sequence <220><223> capable of binding to Ang-2 Peptide <220><22i> misc_ Feature: <222> (26)..(26) <223> Xaa = Fc <400> 32

Met Gly Ala Gin Lys Arg Pro Cys Glu Glu lie Phe Gly Gly Cys Thr 15 10 15 -213- (209)1317281 Description of the Invention Continued:

Tyr Gin Leu Glu Gly Gly Gly Gly Gly Xssi 20 25 <210> 33 <211> 784

<212> DNA <213> artificial sequence <220>

≪ 223 > coding capable of binding to Ang-2 of peptibodies DNA < 400 > 33 atgggtgcac agaaattcaa cccgctggac gaactggaag aaactctgta cgaacagttc actttccagc agctcgaggg tggaggcggt ggggacaaaa ctcacacatg tccaccttgc ccagcacctg aactcctggg gggaccgtca gttttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga tgagctgacc aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag agcctctccc tgtctccggg taaataatgg atcc <210> 34 <211> 768

<212> DNA <213> artificial sequence

60 120 180 240 300 360 420 480 540 600 660 720 780 784

-214· 1317281 (210) "Invention Description Continued Page <220>

≪ 223 > coding capable of binding to Ang-2 of peptibodies DNA < 400 > 34 atgaaattca acccgctgga 4 cgaactggaa gaaactctgt acgaacagtt cactttccag 60 cagctcgagg gtggaggcgg tggggacaaa actcacacat gtccaccttg cccagcacct 120 gaactcctgg ggggaccgtc agttttcctc ttccccccaa aacccaagga caccctcatg 180 atctcccgga cccctgaggt cacatgcgtg gtggtggacg tgagccacga agaccctgag 240 gtcaagttca actggtacgt ggacggcgtg gaggtgcata atgccaagac aaagccgcgg 300 gaggagcagt acaacagcac gtaccgtgtg gtcagcgtcc tcaccgtcct gcaccaggac 360 - tggctgaatg gcaaggagta caagtgcaag gtctccaaca aagccctccc agcccccatc 420 gagaaaacca tctccaaagc caaagggcag ccccgagaac cacaggtgta caccctgccc 480 ccatcccggg atgagctgac caagaaccag gtcagcctga cctgcctggt caaaggcttc 540 tatcccagcg acatcgccgt ggagtgggag agcaatgggc agccggagaa caactacaag 600 accacgcctc ccgtgctgga ctccgacggc tccttcttcc tctacagcaa gctcaccgtg 660 Gacaagagca ggtggcagca ggggaacgtc ttctcatgct ccgtgatgca tgaggctctg 720 cacaaccact acacgcagaa gagcctctcc ctgtctccgg gtaaataa 768 <210> 35 <211> 834 <212> DNA <213> Artificial sequence <220><223> DNA encoding a peptibosome capable of binding to Ang-2 <400> 35 atgaaattca acccgctgga cgaactggaa gaaactctgt acgaacagtt cactttccag 60 cagggatccg gttctgctac tggtggttcc ggctccaccg caagctctgg ttcaggcagt 120 gcgactcatc tcgagggtgg aggcggtggg gacaaaactc acacatgtcc accttgccca 180 gcacctgaac tcctgggggg accgtcagtt ttcctcttcc ccccaaaacc caaggacacc 240 -215- 1317281 (211) Grade invention is described on page ctcatgatct cccggacccc tgaggtcaca tgcgtggtgg tggacgtgag ccacgaagac 300 cctgaggtca agttcaactg gtacgtggac ggcgtggagg tgcataatgc caagacaaag 360 ccgcgggagg agcagtacaa cagcacgtac cgtgtggtca gcgtcctcac cgtcctgcac 420 caggactggc tgaatggcaa ggagtacaag tgcaaggtct ccaacaaagc cctcccagcc 480 cccatcgaga aaaccatctc caaagccaaa gggcagcccc gagaaccaca ggtgtacacc 540 ctgcccccat cccgggatga gctgaccaag aaccaggtca gcctgacctg cctggtcaaa 600 ggcttctatc ccagcgacat cgccgtggag tgggagagca atgggcagcc ggagaacaac 660 tacaagacca cgcct Cccgt gctggactcc gacggctcct tcttcctcta cagcaagctc 720 accgtggaca agagcaggtg gcagcagggg aacgtcttct catgctccgt gatgcatgag 780- gctctgcaca accactacac gcagaagagc ctctccctgt ctccgggtaa ataa 834 <210> 36 <211> 861 <212> DNA <213> artificial sequence <220><223>; the DNA peptibody encoding capable of binding of Ang-2 ... < 400 > 36 atgggtgcac agaaattcaa cccgctggac gaactggaag aaactctgta cgaacagttc 60 actttccagc agggtggtgg tggtggtggc ggtggtaagt tcaacccact ggatgagctg 120 gaagagactc tgtatgaaca gttcactttc cagcaactcg agggtggagg cggtggggac 180 aaaactcaca catgtccacc ttgcccagca cctgaactcc tggggggacc gtcagttttc 240 ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga ggtcacatgc 300 gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta cgtggacggc 360 gtggaggtgc ataatgccaa gacaaagccg cgggaggagc agtacaacag cacgtaccgt 420 gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga gtacaagtgc 480 aaggtctcca acaaagccct cccagccccc atcgagaaaa ccatctccaa agccaaaggg 540 cagccccgag aaccacaggt gt acaccctg cccccatccc gggatgagct gaccaagaac 600 -216- 1317281 _ (212) described on page Continued caggtcagcc invention tgacctgcct ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg 660 gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct ggactccgac 720 ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca gcaggggaac 780 gtcttctcat gctccgtgat gcatgaggct ctgcacaacc actacacgca gaagagcctc 840 tccctgtctc cgggtaaata a 861 <210> 37 <211> 849

<212> DNA <213> artificial sequence <220>

240 aaacccaagg 37 atgaaattca acccgctgga cgaactggaa gaaactctgt acgaacagtt cactttccag 60 cagggtggtg gtggtggcgg tggtaagttc aacccactgg atgagctgga agagactctg 120 tatgaacagt tcactttcca gcaactcgag ggtggaggcg gtggggacaa aactcacaca 180 tgtccacctt gcccagcacc tgaactcctg gggggaccgt cagttttcct cttcccccca; < 223 > peptibody encoding capable of binding of Ang-2 DNA < 400 & gt acaccctcat gatctcccgg acccctgagg tcacatgcgt ggtggtggac 300 gtgagccacg aagaccctga ggtcaagttc aactggtacg tggacggcgt ggaggtgcat 360 aatgccaaga caaagccgcg ggaggagcag tacaacagca cgtaccgtgt ggtcagcgtc 420 ctcaccgtcc tgcaccagga ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac 480 aaagccctcc cagcccccat cgagaaaacc atctccaaag ccaaagggca gccccgagaa 540 ccacaggtgt acaccctgcc cccatcccgg gatgagctga ccaagaacca ggtcagcctg 600 acctgcctgg tcaaaggctt ctatcccagc gacatcgccg tggagtggga gagcaatggg 660 cagccggaga acaactacaa Gaccacgcct cccgtgctgg actccgacgg ctccttcttc 720 ctctacagca agctcaccgt ggacaagagc aggtggcagc aggggaacgt cttctcatgc 780 tccgtgatgc atgaggctct Gcacaaccac tacacgcaga agagcctctc cctgtctccg 840 ggtaaataa 849 •217- 1317281 (213) Description of the Invention Continued: <210> 38 <211> 759

<212> DNA <213> artificial sequence <220>

≪ 223 > peptibody encoding capable of binding of Ang-2 DNA < 400 > 38 atgggtgcac agcaggaaga atgcgaatgg gacccatgga cttgcgaaca catgctcgag 60 ggtggaggcg gtggggacaa aactcacaca tgtccacctt gcccagcacc tgaactcctg 120 gggggaccgt cagttttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 180 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 240 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 300 tacaacagca cgtaccgtgt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 360 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc 420 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 480 gatgagctga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 540 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 600 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 660 aggtggcagc aggggaacgt Cttctcatgc tccgtgatgc atgaggctct gcacaaccac 720 tacacgcaga agagcctctc cctgtctccg ggtaaataa 759

<210> 39 <211> 816 <212> DNA <213> Artificial sequence-218·<220> 1317281 (214) Description of the invention i........ 1 60 120 180 240 300 360 420 480 540 600 660 720 780 816

catgcgtggt ggtggacgtg 39 atgcaggaag aatgcgaatg ggacccatgg acttgcgaac acatgggatc cggttctgct actggtggtt ccggctccac cgcaagctct ggttcaggca gtgcgactca tctcg ^ agggt ggaggcggtg gggacaaaac tcacacatgt ccaccttgcc cagcacctga actcctgggg ggaccgtcag ttttcctctt ccccccaaaa cccaaggaca ccctcatgat ctcccggacc cctgaggtca; < 223> peptide precursor coding capable of binding of Ang-2 DNA < 400 & gt agccacgaag accctgaggt caagttcaac tggtacgtgg acggcgtgga ggtgcataat gccaagacaa agccgcggga ggagcagtac aacagcacgt accgtgtggt cagcgtcctc accgtcctgc accaggactg gctgaatggc aaggagtaca agtgcaaggt ctccaacaaa gccctcccag cccccatcga gaaaaccatc tccaaagcca aagggcagcc ccgagaacca caggtgtaca ccctgccccc atcccgggat gagctgacca agaaccaggt cagcctgacc tgcctggtca aaggcttcta tcccagcgac atcgccgtgg agtgggagag caatgggcag ccggagaaca actacaagac cacgcctccc gtgctggact ccgacggctc cttcttcctc tacagcaagc tcaccgtgga caagagcagg tggcagcagg ggaacgtctt ctcatgctcc gtgatgcatg aggctctgca Caaccactac acgcagaaga gcctctccct gtctccgggt aaataa <210> 40 <211> 867

<212> DNA <213> artificial sequence <220>

≪ 223 > skin member encoding a polypeptide capable binding of Ang-2 DNA < 400 > 40 atgggtgcac agcaggaaga atgcgaatgg gacccatgga cttgcgaaca catgggatcc 60 ggttctgcta ctggtggttc cggctccacc gcaagctctg gttcaggcag tgcgactcat 120 caggaagaat gcgaatggga cccatggact tgcgaacaca tgctcgaggg tggaggcggt 180 ggggacaaaa ctcacacatg tccaccttgc ccagcacctg aactcctggg gggaccgtca 240 gttttcctct Tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 30.0 -219· 1317281 (215) Description of the Invention Continued

acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 360 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 420 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 480 aagtgcaagg tcfccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc 540 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga tgagctgacc 600 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 660 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac · 720 Tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag 780 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 840 agcctctccc tgtctccggg taaataa 867 <210> 41 <211> 774

<212> DNA <213> artificial sequence <220>

<223> DNA encoding a peptibosome capable of binding to Ang-2 <400>

atggacaaaa ctcacacatg tccaccttgc ccagcacctg aactcctggg gggaccgtca 60 gttttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 120 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 180 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 240 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 300 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc 360 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga tgagctgacc 420 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540 tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag 600 -220- 1317281 (216) Grade invention is described on page gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 660 agcctctccc tgtctccggg taaaggtgga ggtggtggtg cacagaaatt caacccgctg 720 gacgagctgg aagagactct gtacgaacag tttacttttc aacagctcga Gtaa 774 <210> 42 ^ <211> 840 <212> DNA <213&g t; artificial sequence

<220>

240 taccgtgtgg 42 atggacaaaa ctcacacatg tccaccttgc ccagcacctg aactcctggg gggaccgtca 60 gttttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 120 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 180 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg; < 223 > peptibody encoding capable of binding of Ang-2 DNA < 400 & gt tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 300 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc 360 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga tgagctgacc 420 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540 tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag 600 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 660 agcctctccc tgtctccggg Taaaggtgga ggtggtggtg cacagggatc cggttctgct 720 actggtggtt ccggctccac cgcaagctct ggttcaggca gtgcgactca taaattcaac 780 ccgctggac g aactggaaga aactctgtac gaacagttca ctttccagca actcgagtaa 840

<210> 43 <211> 858 •221 - 1317281 __ (217) Invention Description

<212> DNA <213> artificial sequence <220>

240 taccgtgtgg 43 atggacaaaa ctcacacatg tccaccttgc ccagcacctg aactcctggg gggaccgtca 60 gttttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 120 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 180 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg; < 223 > peptibody encoding capable of binding of Ang-2 DNA < 400 & gt tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 300 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc 360 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga tgagctgacc 420 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540 tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag 600 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 660 agcctctccc tgtctccggg Taaaggtgga ggtggtggtg cacagaaatt caacccgctg 720 gacgaactgg aagaaactct gtacgaacag ttcactttcc agcagggtgg tggtggtggt 780 ggcggtg Gta agttcaaccc actggatgag ctggaagaga ctctgtatga acagttcact 840 ttccagcaac tcgagtaa 858

<210> 44 <211〉 756

<212> DNA <213> artificial sequence <220>

<223> DNA encoding a peptibosome capable of binding to Ang-2 - 222 - 1317281 (218) Description of the invention Continuation page <400> 44 atggacaaaa ctcacacatg tccaccttgc ccagcacctg aactcctggg gggaccgtca 60 gttttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 120 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 180 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 240 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 300 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc 360 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga tgagctgacc 420 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540 tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag 600 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 660 agcctctccc tgtctccggg taaaggtgga ggtggtggtg cacagcagga agaatgcgaa 720 tgggacccat ggacttgcga acacatgctc gagtaa 756 < 210> 45, < 211> 822 < 212 > DMA <213>man-made sequence

<220>

240 taccgtgtgg 45 atggacaaaa ctcacacatg tccaccttgc ccagcacctg aactcctggg gggaccgtca 60 gttttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 120 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 180 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg; < 223 > peptibody encoding capable of binding of Ang-2 DNA < 400 & gt tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 300 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc 360 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga tgagctgacc 420 -223 - 1317281 _ (219) described on page continued invention 480 540 600 660 720 780 822 aagaaccagg gagtgggaga tccgacggct gggaacgtct agcctctccc actggtggtt tgcgaatggg tcagcctgac gcaatgggca ccttcttcct tctcatgctc Tgtctccggg ccggctccac acccatggac ctgcctggtc gccggagaac ctacagcaag cgtgatgcat taaaggtgga cgcaagctct ttgcgaacac aaaggcttct aactacaaga ctcaccgtgg gaggctctgc ggtggtggtg ggttcaggca atgctcgagt atcccagcga ccacgcctcc acaagagcag acaaccacta cacagggatc gtgcgactca Aa catcgccgtg cgtgctggac gtggcagcag cacgcagaag cggttctgct tcaggaagaa <210> 46 <211> 864

<212> DNA <213> artificial sequence <22〇>

<223> DNA encoding a peptibosome capable of binding to Ang-2 <400> 46 atggacaaaa ctcacacatg tccaccttgc ccagcacctg aactcctggg gggaccgtca 60 gttttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 120 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 180

gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 240 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 300 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc 360 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga tgagctgacc 420 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540 tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag 600 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 660 agcctctccc tgtctccggg taaaggtgga ggtggtggtg cacagcagga agaatgcgaa 720 tgggacccat ggacttgcga acacatggga tccggttctg ctactggtgg ttccggctcc 780 -224- 1317281 (220) the invention described performance page accgcaagct ctggttcagg cagcgcgact catcaggaag aatgcgaatg ggacccatgg 840 acttgcgaac acatgctcga gtaa 864 < 210 > 47 < 211 > 906 '

<212> DNA <213> artificial sequence

≪ 223 > peptibody encoding capable of binding of Ang-2 DNA < 400 > 47 atgggtgcac aggaagaatg cgaatgggac ccatggactt gcgaacacat gggtggtggt 60 ggtggtggcg gtggtaaatt caacccgctg gacgaactgg aagaaactct gtacgaacag 120 ttcactttcc agcagggatc cggttctgct actggtggtt ccggctccac cgcaagctct 180 ggttcaggca gtgcgactca tctcgagggt ggaggcggtg gggacaaaac tcacacatgt 240 ccaccttgcc cagcacctga actcctgggg ggaccgtcag ttttcctctt ccccccaaaa 300 cccaaggaca ccctcatgat ctcccggacc cctgaggtca catgcgtggt ggtggacgtg 360 agccacgaag accctgaggt caagttcaac tggtacgtgg acggcgtgga ggtgcataat 420 gccaagacaa agccgcggga ggagcagtac aacagcacgt accgtgtggt cagcgtcctc 480 accgtcctgc accaggactg gctgaatggc aaggagtaca agtgcaaggt ctccaacaaa 540 gccctcccag cccccatcga gaaaaccatc tccaaagcca aagggcagcc ccgagaacca 600 caggtgtaca ccctgccccc atcccgggat gagctgacca agaaccaggt cagcctgacc 660 tgcctggtca aaggcttcta Tcccagcgac atcgccgtgg agtgggagag caatgggcag 720 ccggagaaca actacaagac cacgcctccc gtgctggact ccgacggctc cttcttcctc 780 tacagcaagc Tcaccgtgga caagagcagg tggcagcagg ggaacgtctt ctcatgctcc 840 gtgatgcatg aggctctgca caaccactac acgcagaaga gcctctccct gtctccgggt 900 aaataa 906

<210> 48 • 225· 1317281 (221) I invention description continuation page <211> 897 <212> DNA <213> artificial sequence <22〇>

240 taccgtgtgg 48 atggacaaaa ctcacacatg tccaccttgc ccagcacctg aactcctggg gggaccgtca 60 gttttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 120 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 180 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg; < 223 > peptibody encoding capable of binding of Ang-2 DNA < 400 & gt tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 300 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc 360 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga tgagctgacc 420 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540 tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag 600 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 660 agcctctccc tgtctccggg Taaaggtgga ggtggtggtg cacagggatc cggttctgct 720 actggtggtt ccggctccac cgcaagctct ggttcaggca gtgcgactca taaattcaac 780 ccgctgga Cg aactggaaga aactctgtac gaacagttca ctttccagca gggtggtggc 840 ggtggtcagg aagaatgcga atgggaccca tggacttgcg aacacatgct cgagtaa 897

<210> 49 <211> 771 <212> DNA <213> Artificial sequence -226· <220> 1317281 (222) Description of the invention

≪ 223 > peptibody encoding capable of binding of Ang-2 DNA < 400 > 49 atgggtgcac agttcgacta ctgcgaaggt gttgaagacc cgttcacttt cggttgcgac 60 aaccacctcg agggtggagg cggtggggac aaaactcaca catgtccacc ttgcccagca 120 cctgaactcc tggggggacc gtcagttttc ctcttccccc caaaacccaa ggacaccctc 180 atgatctccc ggacccctga ggtcacatgc gtggtggtgg acgtgagcca cgaagaccct 240 gaggtcaagt Tcaactggta cgtggacggc gtggaggtgc ataatgccaa gacaaagccg 300 cgggaggagc agtacaacag cacgtaccgt gtggtcagcg tcctcaccgt cctgcaccag 360

gactggctga atggcaagga gtacaagtgc aaggtctcca acaaagccct cccagccccc 420 atcgagaaaa ccatctccaa agccaaaggg cagccccgag aaccacaggt gtacaccctg 480 cccccatccc gggatgagct gaccaagaac caggtcagcc tgacctgcct ggtcaaaggc 540 ttctatccca gcgacatcgc cgtggagtgg gagagcaatg ggcagccgga gaacaactac 600 aagaccacgc ctcccgtgct ggactccgac ggctccttct tcctctacag caagctcacc 660 gtggacaaga gcaggtggca gcaggggaac gtcttctcat gctccgtgat gcatgaggct 720 ctgcacaacc actacacgca gaagagcctc tccctgtctc cgggtaaata a 771 <210> 50 <211> 759

<212> DNA <213> artificial sequence <220>

≪ 223 > peptibody encoding capable of binding of Ang-2 DNA < 400 > 50 atgggtgcac agcagtacgg ttgcgacggt tttctgtacg gttgcatgat caacctcgag 60 ggtggaggcg gtggggacaa aactcacaca tgtccacctt gcccagcacc tgaactcctg 120 gggggaccgt cagttttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 180 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 240 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag. 300 tacaacagca cgtaccgtgt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 360 -227- 1317281 (223) described on page continued invention ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc 420 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 480 gatgagctga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 540 gacatcgccg tg ^ agtggga Gagcaatggg cagccggaga acaactacaa gaccacgcct 600 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 660 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 720 tacacgcaga agagcctctc cctgtctccg ggtaaataa 7 59 <210> 51 <211> 759

<212> DNA <213> artificial sequence <220>

<223> DNA encoding a peptibosome capable of binding to Ang-2

≪ 400 > 51 atgggtgcac agaaacgccc atgcgaagaa atgtggggtg gttgcaacta cgacctcgag 60 ggtggaggcg gtggggacaa aactcacaca tgtccacctt gcccagcacc tgaactcctg 120 gggggaccgt cagttttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 180 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 240 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 300 tacaacagca cgtaccgtgt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 360 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc 420 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 480 gatgagctga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 540 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 600 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 660 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 720 tacacgcaga agagcctctc cctgtctccg ggtaaataa 759 • 228- 1317281 (224 ) Description of the Invention Continued: <210> 52 <211> 759

<212> DNA <213> artificial sequence <220>

aatgccaaga caaagccgcg 52 atgggtgcac agcaccagat ctgcaaatgg gacccgtgga cctgcaaaca ctggctcgag ggtggaggcg gtggggacaa aactcacaca tgtccacctt gcccagcacc tgaactcctg gggggaccgt cagttttcct cttcccccca aaacccaagg acaccctcat gatctcccgg acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc aactggtacg tggacggcgt ggaggtgcat; < 223 > peptibody encoding capable of binding of Ang-2 DNA < 400 & gt ggaggagcag tacaaicagca cgtaccgtgt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc 'cagcccccat cgagaaaacc atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg gatgagctga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac tacacgcaga agagcctctc cctgtctccg ggtaaataa <210> 53 <211> 759

<212> DNA <2i3> artificial sequence <220> 60 120 180 240 300 360 420 480 540 600 660 720 759

•229- 1317281 (225) Description of the Invention Continued

≪ 223 > coding capable of binding to Ang-2 of peptibodies DN A < 400 > 53 atgggtgcac agaaacgtcc atgcgaagaa atcttcggtg gttgcaccta ccagctcgag 60 ggtggaggcg gtggggacaa aactcacaca tgtccacctt gcccagcacc tgaactcctg 120 gggggaccgt cagttttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 180 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 240 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 300 tacaacagca cgtaccgtgt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 360 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat cgagaaaacc 420 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 480 gatgagctga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 540 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 600 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 660 aggtggcagc Aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 720 tacacgcaga agagcctctc cctgtctccg ggtaaataa 759 <210> 54 <211> 25

<212> DNA <213> Artificial sequence <220><223> Oligonucleotide <400> 54 cgqcqcaact atcggtatca agctg 25 <210> 55

<211> 26 <212> DNA <213> Artificial sequence - 230 - 1317281 Description of the invention (226) <220><223> Oligonucleotide <400> 55 26 catgtaccgt aacactgagt ttcgtc <lt;;210> 56 <211> 14 <212>protein <213> artificial sequence

<220><223> Consistent theme <220><221>misc_ feature <222> (7, 12 and)..(14) <223> Xaa means produced from the TN8-IX library Any naturally occurring amino acid. <400> 56

Lys Arg Pro Cys Glu Glu Xaa Trp Gly Gly Cys Xaa Tyr Xaa 15 10 <210> 57 <211> 14 <212> Protein <213> Artificial Sequence <220><223>&TN8- Consistent theme produced in the IX library <220> -231 - 1317281 Description of the invention (227) <221>misc_features<222>(7,12 and)..(14) <223> Xaa Refers to any naturally occurring amino acid. <400> 57

Lys Arg Pro Cys Glu Glu Xaa Phe Gly Gly Cys Xaa Tyr Xaa 5 10

<210> 58 <211> 14 <212> Protein <213> Artificial sequence <220><223> Consistent theme <220><221> misc_ produced from the TN8-IX library Characteristic <222> (1, 2, 3, 5 and).. (13) <223> Xaa means any naturally occurring amino acid.

<400〉58

Xaa Xaa Xaa Cys Xaa Trp Asp Pro Trp Thr Cys Glu Xaa Met 1 5 l〇<210> 59 <211>18 <212>Protein<213> Artificial Sequence 232 - <220> 1317281 Page (228) <223> Consistent theme <220><221> misc_features <222> (3, 8, 10-14 and) (18) <223> Xaa means any naturally occurring amino acid. <400> 59

Trp Ser Xaa Cys Ala Trp Phe Xaa Gly Xaa Xaa Xaa Xaa Xaa Cys Arg 15 10 15

Arg Xaa <210> 60 <211> 227 <212> Protein <213> Artificial sequence <220><223> Human Fc IgG1 <400> 60

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

Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 20 25 30 lie Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 35 40 45

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

His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gin Tyr Asn Ser Thr Tyr 65 70 75 80 -233 - 1317281 _ (229) Description of the Invention Continued

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

Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro lie I90 105 110

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

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

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

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

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

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

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

Pro Gly Lys 225 <210> 61 <211>14 <212> Protein <213> Artificial Sequence <220><223> Consistent Theme Produced From TN8-IX Library - 234 - 1317281 Continued (230) <220><221>misc_features<222> (1-3, 5, 7, 12, 13 and)..(14) <223> Xaa means any naturally occurring Amino acid. <400> 61

Xaa Xaa Xaa Cys Xaa Asp Xaa Tyr Trp Tyr Cys Xaa Xaa Xaa 15 10

<210> 62 <211> 14 <212>protein <213> artificial sequence <220><223> Consistent theme produced from TN8-IX library <220><221>misc_#

<222> (1-3, 5, 7, 12, 13 and).. (14) <223> Xaa means any naturally occurring amino acid. <400> 62

Xaa Xaa Xaa Cys Xaa Asp Xaa Tyr Thr Tyr Cys Xaa Xaa Xaa 1 5 10 <210> 63 <211>14 <212> Protein <213> Artificial sequence 235 - 1317281 Description of the invention (231) <;220><223> Consistent theme <220><221>misc_features<222> (1-3, 5, 7, 12, 13 and) (.14) produced from the TN8-IX library <223> Xaa means any naturally occurring amino acid. <400> 63

Xaa Xaa Xaa Cys Xaa Asp Xaa Phe Trp Tyr Cys Xaa Xaa Xaa <210> 64 <211> 14 <212> Protein <213> Artificial Sequence <220><223> From TN8-IX Library Consistent theme

<220><221>misc_ feature <222> (1-3, 5, 7, 12, 13 and).. (14) <223> Xaa means any naturally occurring amino acid. <400> 64

Xaa Xaa Xaa Cys Xaa Asp Xaa Phe Thr Tyr Cys Xaa Xaa Xaa 1 5 10 <210> 65 <211> 5 236- 1317281 Description of the Invention Continued (232) <212> Protein <213> Artificial Sequence <;220><223> A polypeptide capable of binding to Ang-2 <400> 65

Trp Asp Pro Trp Thr 1 5

<210> 66 <211> 6 <212>Protein <213> Artificial sequence <220><223> A polypeptide capable of binding to Ang-2 <400>

Trp Asp Pro Trp Thr Cys 1 5

<210> 67 <211> 7 <212>Protein<213> Artificial sequence <220><223>>223 capable of binding to Ang-2 <220><221>misc_feature<;222> (2)..(2) -237- 1317281 (233) <223> Xaa is an acidic or neutral polar amino acid residue <400> 67

Cys Xaa Trp Asp Pro Trp Thr 1 , 5

<210> 68 <211> 8 <212> Protein < 213 > Artificial Synthetic Sequence <220><223> 223 <220><221><221> misc_characteristic <221>;222> (2)..(2) <223> Xaa is an acidic or neutral polar amino acid residue <400>

Cys Xaa Trp Asp Pro Trp Thr Cys 1 5 <210> 69 <211>14 <212> Protein <213>Artificial Sequence <220><223> A polypeptide capable of binding to Ang-2 <220> -238 - 1317281 Description of the Invention (234) <221>misc_Characteristics<222> (1, 2 and).. (3) <223> Xaa are each an amino acid residue. <220><221> misc_characteristic <222> (5).. (5) <223> Xaa is an amino acid residue. <220><221> misc_ feature <222>(12)..(12) <223> Xaa lacks or is an amino acid residue. <220><22 1 > misc_feature <222> (13)..(13)

<223> Xaa lacks either a neutral hydrophobic, a neutral polar, or a basic amino acid residue. <220><221> misc_characteristics <222> (14).. (14) <223> Xaa is a neutral hydrophobic or neutral polar amino acid residue. <400> 69

Xaa Xaa Xaa Cyas Xaa Trp Asp Pro Trp Thr Cys Xaa Xaa Xaa 1 5 10 • 239- 1317281 (235) Summary of the Invention Page <210> 70 <211> 20 <212> Protein <213><220><223> A polypeptide capable of binding to Ang-2 <220><221> misc_characteristic <222> (1 and)..(15) <223> Xaa is absent or is an amine group Acid residue. <220><221> misc_features <222> (2 and)..(16) <223> Xaa lacks or is a neutral hydrophobic or neutral polar group. <220><221> misc_characteristic <222> (3-6, 18, 19 and).. (20) <223> Xaa lacks or is an amino acid residue, respectively. <220><22 1 > misc_ feature <222> (8).. (8) <223> Xaa is an amino acid residue. Alkaline amino acid residue

-240- 1317281 Description of the Invention Page (236) <220><221>misc_Features<222> (17)..(17) <223> Xaa lacks or is neutral hydrophobic, neutral Polar amino acid residue. <400> 70

Xaa Xaa Xaa Xaa Xaa Xaa Cys Xaa Trp Asp Pro Trp Thr Cys Xaa Xaa 1 5 10 15

Xaa Xaa Xaa Xaa 20 <210> 71 <211> 10 <212> Protein <213> Artificial sequence <220>

<223> Polypeptide capable of binding to Ang-2 <200><221> misc_characteristic <222> (2)..(2) <223> Xaa is a neutral hydrophobic amino acid residue base. <220><221>misc_ feature <222> (4)..(4) <223>Xaa is a, A, D, or E. -241 - Invention Description Continued

1317281 (237) <220><22 1 > miscj+ sign <222> (6).. (6) <223> Xaa is an acidic amino acid residue. <220><221>misc_ feature <222> (7).. (7) <223> Xaa is an amino acid residue. <220><221> misc_characteristics <222> (8).. (8) <223> Xaa is a neutral hydrophobic, neutral polar or an experimental amino acid residue. <400> 71

Pro Xaa Asp Xaa Leu Xaa Xaa Xaa Leu 1 5 <210> 72 <211> 22 <212>Protein<213> Artificial sequence <220><223> A polypeptide capable of binding to Ang-2 ;220><221>misc characteristic-242- 1317281 Description of the Invention (238) <222> (1,4 and) (20) <223> Xaa lacks or is an amino acid residue. <220><221> misc_characteristic <222> (2, 15, 16 and)..(21) <223> Xaa lacks or is a neutral polar, acidic or atrial amino acid Residues.

<220><221> misc_characteristic <222> (3, 17 and)..(18) <223> Xaa is absent, or is a neutral hydrophobic or neutral polar amino acid residue . <220><221> misc_characteristic <222> (6).. (6) <223> Xaa is a neutral hydrophobic amino acid residue. <220><221>misc_feature<222> (8)..(8) <223> Xaa is a A, D, or E 〇 <220><221> misc_ feature <222> (10)..(10) -243 - 1317281 Description of the Invention (239) <223> Xaa is an acidic amino acid residue. <220><221> misc_characteristic <222> (11).. (11) <223> Xaa is an amino acid residue. <220>

<221>misc_ feature <222> (12).. (12) <223> Xaa is a neutral hydrophobic, neutral polar or an experimental amino acid residue. <220><221> misc_characteristic <222> (19 and).. (22) <223> Xaa lacks or a neutral hydrophobic, neutral polar or basic amino acid residue.

<400> 72

Xaa Xaa Xaa Xaa Pro Xaa Asp Xaa Leu Xaa Xaa Xaa Leu Tyr Xaa Xaa 1 5 10 15

Xaa Xaa Xaa Xaa Xaa Xaa 20 <210> 73 <211> 8 <212> Protein <213> Artificial sequence 244 - 1317281 Description of the invention (240) <220><223> -2 bound polypeptide <220><221> misc_characteristic <222> (3).. (3) <223> Xaa is a neutral polar amino acid residue. <220>

<22 1 > misc_ feature <222> (4).. (4) <223> Xaa is an acidic amino acid residue. <220><221> misc_characteristic <222> (5).. (5) <223> Xaa is a neutral polar or acidic amino acid residue. <220>

<221>misc_ feature <222> (6 and).. (7) <223> Xaa is a neutral hydrophobic amino acid residue. <400> 73

Arg Pro Xaa Xaa Xaa Xaa Xaa Gly 1 5 <210> 74 <211> 20 <212> Protein-245 - 1317281 (241) Description of the Invention Continued <213> Artificial Order 歹|J <220><223> A polypeptide capable of binding to Ang-2 <220><221> misc_features <222> (1, 2, 4, 13, 14, 19 and) (20) <223> Xaa is a neutral hydrophobic or neutral polar amino acid residue.

<220><221> misc_characteristic <222> (3, 9 and) (17) <223> Xaa is a neutral polar or acidic amino acid residue. <220><221> misc_characteristic <222> (7, 15 and).. (16) <223> Xaa is a neutral polar amino acid residue.

<220><221> misc_characteristic <222> (8).. (8) <223> Xaa is an acidic amino acid residue. <220><221> misc_characteristic <222> (10 and)..(11) -246 - 1317281 Description of the Invention (242) <223> Xaa is a neutral hydrophobic amino acid Residues. <220><221> misc_ feature <222>(18)..(18) <223> Xaa is a neutral hydrophobic or basic amino acid residue. <400> 74

Xaa Xaa Xaa Xaa Arg Pro Xaa Xaa Xaa Xaa Xaa Gly Xaa Xaa Xaa Xaa 1 5 10 15

Xaa Xaa Xaa Xaa 20 <210> 75 <211> 13 <212> Protein < 213 > Artificial Synthetic Sequence <220><223><223> Touching polypeptide bound to Ang-2 <220> 221 > misc_characteristic <222> (2).. (2) <223> Xaa is an acidic amino acid residue. <220><221> misc_characteristic <222> (4).. (4) <223> Xaa is a neutral hydrophobic amino acid residue. -247- Description of the Invention Continued

1317281 (243) <220><22 1 > misc_ feature <222> (5)..(5) <223> Xaa is E, D, or Q. <220><221> misc_characteristic <222> (10).. (10) <223> Xaa is a neutral hydrophobic or neutral polar amino acid residue. <220><221> misc_ feature <222>(13)..(13) <223> Xaa is an acidic residue. <400> 75

Cys Xaa Gly Xaa Xaa Asp Pro Phe Thr Xaa Gly Cys Xaa 1 5 l〇<210> 76 <211> 20 <212> Protein <213>Artificial Sequence <220><223> -2 bound polypeptide <400> 76

Pro He Arg Gin Glu Glu Cys Asp Trp Asp Pro Trp Thr Cys Glu His 15 10 15 -248- 1317281 _ (244) Description of the Invention Continued

<212&gt

Thr Asn lie Gin Glu Glu Cys Glu Trp Asp Pro Trp Thr Cys Asp His 15 10 15

<220&gt

Trp Tyr Glu Gin Asp Ala Cys Glu Trp Asp Pro Trp Thr Cys Glu His 15 10 15

Met Ala Glu Val 20 <210〉 79

249-(245)1317281 Description of the Invention Continued <211> 20 <212> Protein <213> Artificial Sequence <220>'<223> A polypeptide capable of binding to Ang-2 <400>

Asn Arg Leu Gin Glu Val Cys Glu Trp Asp Pro Trp Thr Cys Glu His 1 5 10 15

<220&gt

Ala Ala Thr Gin Glu Glu Cvs Glu Trp Asp Pro Trp Thr Cys Glu His 15 10 15

Met Pro Arg Ser 20 <210> 81 <211> 20 <212> Protein <213> Artificial sequence 250-(246) 1317281 Description of the invention Continued page <220> <223> Compatible with Ang-2 Bound polypeptide <400> 81

Leu Arg His Gin Glu Gly Cys Glu Trp Asp Pro Trp Thr Cys Glu His 1 . ; 5 10 15

Met Phe Asp Trp .20

<210> 82 <211> 20 <212> Protein <213> Artificial sequence <220><223> A polypeptide capable of binding to Ang-2 <400>

Val Pro Arg Gin Lys Asp Cys Glu Trp Asp Pro Trp Thr Cys Glu His 15 10 15

Met Tyr Val Gly 20

<210> 83 <211> 20 <212> Protein <213> Artificial sequence <220><223> A polypeptide capable of binding to Ang-2 <400>

Ser lie Ser His Glu Glu Cys Glu Trp Asp Pro Trp Thr Cys Glu His 15 10 15 -251 - 1317281 Description of the Invention Continued Page (247)

Met Gin Val Gly . 20 <210> 84 <211> 20 ' <212> Protein <213> Artificial Sequence

<220><223> A polypeptide capable of binding to Ang-2 <400> 84

Trp Ala Ala Gin Glu Glu Cys Glu Trp Asp Pro Trp Thr Cys Glu His 15 10 15

Met Gly Arg Met 20 <210> 85

<211> 20 <212> Protein <213> Artificial sequence <220><223> can be combined with Ang-2 <400>

Thr Trp Pro Gin Asp Lys Cys Glu Trp Asp Pro Trp Thr Cys Glu His 15 10 15

Met Gly Ser Thr 20 <210> 86 252 - Summary of Invention Continuation 1137281 (248) <211> 20 <212> Protein. <213> Artificial sequence <220>'<223> can be combined with Ang -2 bound polypeptide <400> 86

Gly His Ser Gin Glu Glu Cys Gly Trp Asp Pro Trp Thr Cys Glu His 15 10 15

Met Gly Thr Ser 20 <210> 87 <211> 20 <212>protein <213> artificial sequence <220>

<223> A polypeptide capable of binding to Ang-2 <400> 87

Gin His Trp Gin Glu Glu Cys Glu Trp Asp Pro Trp Thr Cys Asp His 15 10 15

Met Pro Ser Lys 20 <210> 88 <211> 20 <212>Protein<213> Artificial Sequence 253 - 1317281 (249) ΓInvention Description Continued <220> <223> 2 bound polypeptide <400> 88

Asn Val Arg Gin Glu Lys Cys Glu Trp Asp Pro Trp Thr Cys Glu His 1 ...5 10 15

Met Pro Val Arg 20 <210> 89 <211> 20 <212> Protein <213> Artificial sequence <220><223> A polypeptide capable of binding to Ang-2 <400>

Lys Ser Gly Gin Val Glu Cys Asn Trp Asp Pro Trp Thr Cys Glu His 15 10 15

Met Pro Arg Asn 20 <210> 90 <211> 20 <212> Protein <213> Artificial sequence <220><223> A polypeptide capable of binding to Ang-2 <400>

Val Lys Thr Gin Glu His Cys Asp Trp Asp Pro Trp Thr Cys Glu His 15 10 15 -254- 1317281 (250)

Met Arg Glu Trp 20 <210> 91 <211> 20 <212> Protein <213> Artificial sequence <220><223> A polypeptide capable of binding to Ang-2

<400> 91

Ala Trp Gly Gin Glu Gly Cys Asp Trp Asp Pro Trp Thr Cys Glu His 15 10 15

Met Leu Pro Met 20 <210> 92 <211> 20 <212>protein <213> artificial sequence

<220><223> A polypeptide capable of binding to Ang-2 <400> 92

Pro Val Asn Gin Glu Asp Cys Glu Trp Asp Pro Trp Thr Cys Glu His 15 l〇 15

Met Pro Pro Met 20 <210> 93 • 255-(251) 13172281 Description of the Invention Continued <211> 20 <212> Protein <213> Artificial Sequence <220>'<223> can be combined with Ang -2 bound polypeptide <400> 93

Arg Ala Pro Gin Glu Asp Cys Glu Trp Asp Pro Trp Thr Cys Ala His 1 5 10 15

Met Asp lie Lys 20 <210> 94 <211> 20 <212> Protein <213> Artificial sequence <220><223> Multi-limb capable of binding to Ang-2

<400> 94

His Gly Gin Asn Met Glu Cys Glu Trp Asp Pro Trp Thr Cys G丄u His 1 5 10* 15

Met Phe Arg Tyr 20 <210> 95 <211> 20 <212> Protein <213> Artificial sequence -256 - 1317281 (252) Description of the Invention Page <220><223> 2 bound polypeptide <400> 95

Pro Arg Leu Gin Gla Glu Cys Val Trp Asp Pro Trp Thr Cys Glu His 1,5 10 15

Met Pro Leu Arg 20 <210> 96 <211> 20 <212> Protein <213> Artificial sequence <220><223> Polymorphism capable of binding to Ang-2 <400>

Arg Thr Thr Gin Glu Lvs Cys Glu Trp Asp Pro Trp Thr Cys Glu His 1 5 10 15

Met Glu Ser Gin 20 <210> 97 <211> 20 <212> Protein <213> Artificial sequence <220><223> A polypeptide capable of binding to Ang-2 <400>

Gin Thr Ser Gin Glu Asp Cys Val Trp Asp Pro Trp Thr Cys Asp His 15 10 15 -257- 1317281 __ (253) Description of the Invention Continued

Met Val Ser Ser 20 <210> 98 <211> 20 <212> Protein <213> Artificial Sequence <220>

<223> A polypeptide capable of binding to Ang-2 <400> 98

Gin Val He Gly Arg Pro Cys Gin Trp Asp Pro Trp Thr Cys Glu His 15 10 15

Leu Glu Gly Leu 20 <210> 99' <211> 20 <212>protein <213> artificial sequence

<220><223> A polypeptide capable of binding to Ang-2 <400>·*99 Τγό Ala Gin Gin Glu Glu Cys Ala Trp Asp Pro Trp Thr Cys Asp His 1 5 10 15

Met Val Gly Leu 20 <210> 100 -258- Invention Description Page 1327281 (254) <211> 20 <212> Protein <213> Man-made Order. Column <220><223> Polypeptide combined with Ang-2 <400> 100

Leu Pro Gly Gin Glu Asp Cys Glu Trp Asp Pro Trp Thr Cys Glu His 15 10 15

<210&gt

Pro Met Asn Gin Val Glu Cys Asp Trp Asp Pro Trp Thr Cys Glu His 15 10 15

Met Pro Arg Ser 20 <210> 102 <211> 20 <212>Protein<213> Artificial sequence 259 - Description of the invention continued pp. 1317281 (4) <220><223> capable of binding to Ang-2 Peptide-<400> 102

Phe Gly Trp Ser His Gly Cys Glu Trp Asp Pro Trp Thr Cys Glu His 1,5 10 15

<210&gt

Lys Ser Thr Gin Asp Asp Cys Asp Trp Asp Pro Trp Thr Cys Glu His 15 10 15

Met Val Gly Pro 20 <210> 104 <211> 20 <212> Protein <213> Artificial sequence <220><223> A polypeptide capable of binding to Ang-2 <400>

Gly Pro Arg lie Ser Thr Cys Gin Trp Asp Pro Trp Thr Cys Glu His 1 5 10 15 -260- 1317281 (256)

Met Asp Gin Leu 20 <210> 105 <211> 20 <212> Protein <213> Artificial sequence <220><223> A polypeptide capable of binding to Ang-2

<400> 105

Ser Thr He Gly Asp Met Cys Glu Trp Asp Pro Trp Thr Cys Ala His 15 l〇 15

Met Gin Val Asp 20 <210> 106 <211> 20 <212>protein <213> artificial sequence

<220><223> A polypeptide capable of binding to Ang-2 <400> 106

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

Leu Gin Gly Trp 20 &lt;210&gt; 107 •261 · 1317281 (257) Description of the Invention Continued <211> 20 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;'&lt;223&gt; can be combined with Ang -2 bound polypeptide &lt;400&gt; 107

Val Leu Gly Gly Gin Gly Cys Gin Trp Asp Pro Trp Thr Cys Ser His 1 5 10 15

Leu Glu Asp Gly 20 &lt;210> 108 &lt;211&gt; 20 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt;

Thr Thr lie Gly Ser Met Cys Glu Trp Asp Pro Trp Thr Cys Ala His 1 5 10 15

&lt;210&gt Bound polypeptide &lt;400&gt; 109

Thr Lys Gly Lys Ser Val Cys Gin Trp Asp Pro Trp Thr Cys Ser His 1,5 10 15

&lt;220&gt

Thr Thr lie Gly Ser Met Cys Gin Trp Asp Pro Trp Thr Cys Ala His 15 10 15

&lt;210&gt

Trp Val Asn Glu Val Val Cys Glu Trp Asp Pro Trp Thr Cys Asn His 15 10 15 -263· 1317281 _ (259) Description of the Invention Continued

Trp Asp Thr Pro 20 &lt;210&gt; 112 &lt;211&gt; 20 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;

&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt;

Val Val Gin Val Gly Met Cys Gin Trp Asp Pro Trp Thr Cys Lys His 15 10 15

Met Arg Leu G丄n 20 &lt;210&gt; 113 &lt;211&gt; 20 &lt;212&gt;protein &lt;213&gt; artificial sequence

&lt;220&gt;&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt; 113

Ala Val Gly Ser Gin Thr Cys Glu Trp Asp Pro Trp Thr Cys Ala His 15 10 15

Leu Val Glu Val 20 &lt;210&gt; 114 264- 1317281 (260) "Invention Description Continued Continuation Page &lt;211&gt; 20 - &lt;212&gt; Protein &lt;213&gt; Man-made Sequence &lt;220&gt;&lt;223&gt; -2 bound polypeptide &lt;400&gt; 114

Gin Gly Met Lys Met Phe Cys Glu Trp Asp Pro Trp Thr Cys Ala His 15 10 15

Lie Val Tyr Arg 20 &lt;210&gt; 115 &lt;211&gt; 20 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt;

Thr Thr lie Gly Ser Met Cys Gin Trp Asp Pro Trp Thr Cys Glu His 1 5 l〇 . 15

Met Gin Gly Gly 20 &lt;210&gt; 116 &lt;211&gt; 20 &lt;212&gt;Protein&lt;213&gt; Artificial sequence 265- 1317281 _ (261) I invention description continuation page &lt;220&gt;&lt;223&gt; -2 bound polypeptide &lt;400&gt; 116

Thr Ser Gin Arg Val Gly Cys Glu Trp Asp Pro Trp Thr Cys Gin His 1.. , 5 10 15

Leu Thr Tyr Thr 20 &lt;210&gt; 117

&lt;211&gt; 20 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt;

Gin Trp Ser Trp Pro .Pro Cys Glu Trp Asp Pro Trp Thr Cys Gin Thr 1 5 10 15

Val Trp Pro Ser 20

&lt;210&gt; 118 &lt;211&gt; 20 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt;

Gly Thr Ser Pro Ser Phe Cys Gin Trp Asp Pro Trp Thr Cys Ser His 15 10 15 -266- 1317281 _ (262) Description of the Invention Continued

Met Val Gin Gly 20 &lt;210&gt; 119 &lt;211&gt; 22 * &lt;212&gt;protein &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt;

Gin Asn Tyr Lys Pro Leu Asp Glu Leu Asp Ala Thr Leu Tyr Glu His 1 5 10 15

Phe lie Phe His Tyr Thr 20 &lt;210&gt; 120 &lt;211&gt; 22 &lt;212&gt;protein &lt;213&gt; artificial sequence

&lt;220&gt;&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt;

Leu Asn Phe Thr Pro Leu Asp Glu Leu Glu Gin Thr Leu Tyr Glu Gin 15 10 15

Trn Thr Leu Gin Gin Ser &quot; 20 &lt;210&gt; 121 -267· Description of the Invention Continued page 1317281 (263) &lt;211&gt; 22 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt; ' &lt;223&gt a polypeptide capable of binding to Ang-2 &lt;400&gt; 121

Thr Lys Phe Asn Pro Leu Asp Glu Leu Glu Gin Thr Leu Tyr Glu Gin

&lt;220&gt; Protein &lt

&lt;400&gt; 122

Val Lys Phe Lys Pro Leu Asp Ma Leu Glu Gin Thr Leu Tyr Glu His 1 5 ' 10 15

Trp Met Phe Gin Gin Ala 20 &lt;210&gt; 123 &lt;211&gt; 22 &lt;212&gt;Protein&lt;213&gt; Artificial sequence 268-1317281 (264) Illustrated continuation page &lt;220&gt;&lt;223&gt; -2 bound polypeptide &lt;400&gt; 123

Val Lys Tyr Lys Pro Leu Asp Glu Leu Asp Glu lie Leu Tyr Glu Gin 1,5 10 15

Gin Thr Phe Gin Glu Arg 20 &lt;210&gt; 124 &lt;211&gt; 22 &lt;212&gt;Protein&lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Multiple jump capable of binding to Ang-2 &lt;400&gt; 124

Thr Asn Phe Met Pro Met Asp Asp Leu Glu Gin Arg Leu Tyr Glu Gin 15 10 15

Phe lie Leu Gin G丄n Gly 20

&lt;210&gt; 125 &lt;211&gt;22 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt;

Ser Lys Phe Lys Pro Leu Asp Glu Leu Glu Gin Thr Leu Tyr Glu Gin 15 10 15 269- 1317281 (265)

Trp Thr Leu Gin His Ala 20 Description of the Invention Continuation page &lt;210&gt; 126 &lt;211&gt; 22 , &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt; 126 Gin Lys Phe Gin Pro Leu Asp Glu Leu Glu Gin Thr Leu Tyr Glu Gin 1 5 10 15 Phe Met Leu Gin Gin Ala 20 &lt;210&gt; 127 &lt;211&gt; 22 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Polypeptide capable of binding to Ang-2 &lt;400&gt; 127 Gin Asn Phe Lys Pro Met Asp Glu Leu Glu Asp Thr Leu Tyr Lys Gin 1 5 10 15

Phe Leu Phe Gin His Ser 20 &lt;210&gt; 128 -270- Description of the Invention Continued page 1317281 (266) &lt;211&gt; 22 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt; * &lt;223&gt; Polypeptide bound to Ang-2 &lt;400&gt; 128

Tyr Lys Phe Thr Pro Leu Asp Asp Leu Glu Gin Thr. Leu Tyr Glu Gin 1 - 5 * ..10 15

Trp Thr Leu Gin His Val 20 &lt;210&gt; 129 &lt;211&gt; 22 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt;

Gin Glu Tyr Glu Pro Leu Asp Glu Leu Asp Glu Thr Leu Tyr Asn Gin 1 5 10* 15

Trp Met Phe His Gin Arg 20 &lt;210&gt; 130 &lt;211&gt; 22 &lt;212&gt;Protein &lt;213&gt; Artificial Sequence 271 - Invention Description Continued 13172281 (267) &lt;220&gt;&lt;223&gt; -2 bound polypeptide &lt;400&gt; 130

Ser Asn Phe Met Pro Leu Asp Glu Leu Glu Gin Thr Leu Tyr Glu Gin 1 5 10 15

Phe Met Leu Gin His Gin 20 &lt;210&gt; 131

&lt;211&gt; 22 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt;

Gin Lys Tyr Gin Pro Leu Asp Glu Leu Asp Lys Thr Leu Tyr Asp Gin 15 10 15

Phe Met Leu Gin Gin Gly 20

&lt;210&gt; 132 &lt;211&gt; 22 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt;

Gin Lys Phe Gin Pro Leu Asp Glu Leu Glu Glu Thr Leu Tyr Lys Gin 15 10 15 -272· 1317281 Announcement of Invention Page (268)

Trp Thr Leu Gin Gin Arg 20 &lt;210&gt; 133 &lt;211&gt; 22 &lt;212&gt;protein &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt;

Val Lys Tyr Lys Pro Leu Asp Glu Leu Asp Glu Trp Leu Tyr His Gin 15 10 15

Phe Thr Leu His His Gin 20 &lt;210&gt; 134 &lt;211&gt; 22 &lt;212&gt;protein &lt;213&gt; artificial sequence

&lt;220&gt;&lt;223&gt; can be combined with Ang-2 &lt;400&gt; 134

Gin Lys Phe Met Pro Leu Asp Glu Leu Asp Glu lie Leu Tyr Glu Gin 15 10 15

Phe Met Phe Gin Gin Ser 20 &lt;210&gt; 135 -273 - 1317281 (269) &lt;211&gt; 22 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;'&lt;223&gt; capable of interacting with Ang-2 Bound polypeptide - &lt;400&gt; 135 Gin Thr Phe Gin Pro Leu Asp Asp Leu Glu Glu Tyr Leu Tyr 1 5 10 Trp worker Arg Arg Tyr His 20 &lt;210&gt; 136 &lt;211&gt; 22 &lt;212&gt; Protein &lt;;213&gt; artificial sequence^ &lt;220&gt;&lt;223&gt; polypeptide capable of binding to Ang-2 &lt;400&gt; 136 Glu Asp Tyr Met Pro Leu Asp Ala Leu Asp Ala Gin Leu Tyr 15 10 Phe lie Leu Leu His Gly 20 &lt;210&gt; 137 &lt;211&gt; 22 &lt;212&gt;Protein&lt;213&gt; Artificial Sequence Invention Description Continued

Glu Gin 15

Glu Gin 15 274- SYNTHESIS PROCEDURE CONTINUED 1317281 (270) &lt;220&gt;&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt;

His Thr Phe Gin Pro Leu Asp Glu Leu Glu Glu Thr Leu Tyr Tyr Gin 1,5 10 15

Trp Leu Tyr Asp Gin Leu 20 &lt;210&gt; 138

&lt;211&gt; 22 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt;

Tyr Lys Phe Asn Pro Met Asp Glu Leu Glu Gin Thr Leu Tyr Glu Glu 15 10 15

Phe Leu Phe Gin His Ala 20

&lt;210&gt; 139 &lt;211&gt; 22 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;4〇0&gt; 139

Thr Asn Tyr Lys Pro Leu Asp Glu Leu Asp Ala Thr Leu Tyr Glu His 15 10 15 • 275 - 1317281 (271)

Trp lie Leu Gin His Ser 20 &lt;210&gt; 140 &lt;211&gt; 22 * &lt;212&gt; Protein &lt;213&gt; Artificial sequence 5 &lt;220&gt;&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt; 140 Gin Lys Phe Lys Pro Leu Asp Glu Leu Glu Gin Thr Leu Tyr 15 10 Trp Thr Leu Gin Gin Arg 20 &lt;210&gt; 141 &lt;211&gt; 22 &lt;212&gt; Protein &lt;213&gt; Man-made Sequence &lt;220&gt;&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt; 141 Thr Lys Phe Gin Pro Leu Asp Glu Leu Asp Gin Thr Leu Tyr 15 10 Trp Thr Leu Gin Gin Arg 20 &lt;210&gt; 142 Description of the Invention

Glu Gin15

Glu Gin15 -276- 1317281 _ (272) Description of the Invention Continued &lt;211&gt; 22 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt; 1 &lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt; 142

Thr Asn Phe Gin Pro Leu Asp Glu Leu Asp Gin Thr Leu Tyr Glu Gin 15 10 15

Trp Thr Leu Gin Gin Arg 20 &lt;210&gt; 143 &lt;211&gt; 20 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; A polypeptide capable of binding to Ang-2

&lt;400&gt; 143

Ala Gly Gly Met Arg Pro Tyr Asp Gly Met Leu Gly Trp Pro Asn Tyr 15 10 15

Asp Val G丄n Ala 20 &lt;210&gt; 144 &lt;211&gt; 20 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence 277 - Invention Description Continued Page 1327881 (273) &lt;220&gt;&lt;223&gt; -2 bound polypeptide &lt;400&gt; 144

Gin Thr Trp Asp Asp Pro Cys Met His He Leu Gly Pro Val Thr Trp 1 5 10 15

Arg Arg Cys lie 20 &lt;210&gt; 145 &lt;211&gt; 20 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt;

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

Gin Arg worker le Val 20 &lt;210&gt; 146 &lt;211&gt; 20 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt;

Ser Gly Gin Leu Arg Pro Cys Glu Glu lie Phe Gly Cys Gly Thr Gin 1 5 10 15 -278· 1317281 (274)

Asn Leu Ala Leu 20 &lt;210&gt; 147 &lt;211&gt; 20 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt;

Phe Gly Asp Lys Arg Pro Leu Glu Cys Met Phe Gly Gly 15 10

Leu Cys Pro Arg 20 &lt;210&gt; 148 &lt;211&gt; 20 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt;

Gly Gin Asp Leu Arg Pro Cys Glu Asp Met Phe Gly Cys 15 10

Asp Trp Tyr Gly 20 &lt;210&gt; 149

Pro lie Gin 15

Gly Thr Lys 15 279- 1317281 (275) Summary of Invention Continuation Page &lt;211&gt; 20 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; A polypeptide capable of binding to Ang_2 &lt;400&gt;

Gly Phe Glu Tyr Cys Asp Gly Met Glu Asp Pro Phe Thr Phe Gly Cys 1 5 10 ' 15

&lt;210&gt

&lt;400〉 150

Lys Leu Glu Tyr Cys Asp Gly Met Glu Asp Pro Phe Thr Gin Gly Cys 15 10 15

Asp Asn Gin Ser 20 &lt;210&gt; 151 &lt;211&gt; 20 &lt;212&gt; Protein &lt;n3&gt; Artificial Sequence -280 - 1317281 Description of the Invention (276) &lt;220&gt;&lt;223&gt; 2 bound polypeptide &lt;400&gt; 151

Gly Cys 15

Leu Gin Glu Trp Cys Glu Gly Val Glu Asp Pro Phe Thr Phe 1 , 5 10

Glu Lys Gin Arg 20 &lt;210&gt; 152 &lt;211&gt; 20 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt;

Gly Cys 15

Ala Gin Asp Tyr Cys G丄u Gly Met Glu Asp Pro Phe Thr Phe 15 10

Glu Met Gin Lys 20 &lt;210&gt; 153 &lt;211&gt; 20 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt;

Gly Cys 15

Leu Leu Asp Tyr Cys Glu Gly Val Gin Asp Pro Phe Thr Phe 15 10 •281 - 1317281 (277)

Glu Asn Leu Asp 20 &lt;210&gt; 154 &lt;211&gt; 20 &lt;212&gt; Protein &lt;n3&gt; Artificial Sequence &lt;220&gt;

&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400> 154

His Gin Glu Tyr Cys Glu Gly Met Glu Asp Pro Phe Thr ?he Gly Cys 15 10 15

Glu Tyr Gin Gly 20 &lt;210&gt; 155 &lt;211&gt; 20 &lt;212&gt;protein &lt;213&gt; artificial sequence

&lt;220&gt;&lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt;

Met Leu Asp Tyr Cys Glu Gly Met Asp Asp Pro Phe Thr Phe Gly Cys 1 * 5 10 15

Asp Lys Gin Met 20 &lt;210&gt; 156 •282, 1317281 _ (278) Description of the Invention Continued

&lt;211> 20 &lt;212&gt; Protein &lt;213&gt; Artificial sequence J &lt;220&gt; ί &lt;223&gt; A polypeptide capable of binding to Ang-2 &lt;400&gt;

Leu Gin Asp Tyr Cys Glu Gly Val Glu Asp Pro Phe Thr Phe Gly Cys 1 5 10 15

Glu Asn Gin Arg 20 &lt;210&gt; 157 &lt;211&gt; 20 &lt;212&gt;_protein &lt;213&gt; artificial sequence &lt;220&gt;&lt;223&gt; polypeptide capable of binding to Ang-2 &lt;400&gt;

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

Glu Lys Gin Arg 20 &lt;210&gt; 158 &lt;211&gt; 20 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence - 283 -

1317281 (279) &lt;220&gt;&lt;223> polypeptide capable of binding to Ang-2 &lt;220&gt;&lt;221&gt; misc_feature &lt;222&gt; (1)..(1) &lt;223&gt; Xaa is absent Or a neutral hydrophobic, neutral polar or basic amino acid residue. &lt;220&gt;

&lt;22 1&gt; misc_ feature &lt;222&gt; (2, 4, 14).. (19) &lt;223&gt; Xaa neutral hydrophobic or neutral polar amino acid residue. &lt;220&gt;&lt;221&gt; misc_characteristic &lt;222&gt; (3, 6).. (17) &lt;223&gt; Xaa is an acidic amino acid residue.

&lt;220&gt;&lt;221&gt; misc_ feature &lt;222>(8)..(8) &lt;223> Xaa is a neutral hydrophobic amino acid residue. &lt;220&gt;&lt;221&gt; misc_features &lt;222&gt; (9)..(9) &lt;223&gt; Xaa is E, D, or Q. -284- 奋丽_闻1317281 (280) &lt;220&gt;&lt;221&gt;misc_feature&lt;222&gt; (18)..(18) &lt;223&gt; Xaa is neutral hydrophobic, neutral polarity or testability Amino acid residue. &lt;220&gt;&lt;221&gt; misc_characteristic &lt;222&gt; (20).. (20) &lt;223&gt; Xaa is absent or an amino acid residue. &lt;400&gt; 158

Xaa Xaa Xaa Xaa Cys Xaa Glv Xaa Xaa Asp Pro Phe Thr Xaa Gly Cys 1 5 * 10 15

Xaa Xaa Xaa Xaa 20

&lt;210&gt; 159 &lt;211&gt; 60 &lt;212&gt; DNA &lt; 213 &gt; artificial sequence&lt;220&gt;&lt;223&gt; </ br> DNA capable of binding to Ang-2 peptide &lt;400&gt; 159 ccg.atccgtc Aggaagaatg cgactgggac ccgtggacct gcgaacacat gtgggaagtt 60

&lt;210&gt; 160 &lt;211&gt; 60 &lt;212&gt; DNA -285 - 1317281 (281) &lt;213&gt; Artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400> 160 accaacatcc aggaagaatg cgaatgggac ccgtggacct gcgaccacat gccgggtaaa 60 &lt;210&gt; 161 &lt;211&gt; 60 &lt;212&gt; Dm &lt;213&gt;;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 161 tggtacgaac aggacgcttg cgaatgggac ccgtggacct gcgaacacat ggctgaagtt 60 &lt;210&gt; 162 &lt;211&gt; 60

&lt;212> DNA &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 162 aaccgtctgc aggaagtttg cgaatgggac ccgtggacct gcgaacacat ggaaaacgtt 60 &lt;210> 163

&lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; artificial sequence -286 - 1317281 (282) &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 163 gctgctaccc aggaagaatg cgaatgggac ccgtggacct gcgaacacat gccgcgttcc &lt;210&gt; 164 &lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; artificial sequence&lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 164 ctgcgtcacc aggaaggttg cgaatgggac ccgtggacct gcgaacacat gttcgactgg &lt;210&gt; 165 &lt;211&gt; 60

&lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 165 gttccgcgtc agaaagactg cgaatgggac ccgtggacct gcgaacacat gtacgttggt 60 60

60 &lt;210&gt; 166 &lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; artificial sequence -287 - 1317281 issued by month description i (283) &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 166 tgggctgctc aggaagaatg cgaatgggat ccgtggactt gcgaacacat gggtcgtatg 60 &lt;210&gt;

&lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt;: Artificial sequence &lt;220&gt;&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2: &lt;400&gt; 167 acttggccgc aggacaaatg cgaatgggat ccgtggactt gcgaacacat gggttctact 60 &lt;210&gt; 168 &lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 168 ggtcactccc aggaagaatg cggttgggac ccgtggacct gcgaacacat gggtacgtcc 60 &lt;210&gt;

&lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt; •288 - 1317281 (284)

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 169 cagcactggc aggaagaatg cgaatgggac ccgtggacct gcgaccacat gccgtccaaa 60 &lt;210&gt; 170

&lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; A DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 170 aacgttcgtc aggaaaaatg cgaatgggac ccgtggacct gcgaacacat gccggttcgt 60 &lt;210&gt; 171 &lt;211&gt;

&lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 171 aaatccggtc aggttgaatg caactgggac ccgtggacct gcgaacacat gccgcgtaac 60 &lt;210&gt;

&lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 1317281 (285) I "SO® &lt;400&gt; 172

Gttaaaaccc aggaacactg cgactgggac ccgtggacct gcgaacacat gcgtgaatgg SO &lt;210&gt; 173

&lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 60

&lt;400&gt; 173 gcttggggtc aggaaggttg cgactgggac ccgtggacct gcgaacacat gctgccgatg &lt;210&gt; 174

&lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt;

Ccggttaacc aggaagactg cgaatgggac ccgtggacct gcgaacacat gccgccgatg 60 &lt;210&gt; 175 &lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; A DNA encoding a binding to Ang-2 &lt;400&gt; 175 cgtgctccgc aggaagactg cgaatgggac ccgtggacct gcgctcacat ggacatcaaa 60 -290· 1317281 (286) &lt;210&gt; 176 &lt;211&gt; 60 &lt;212&gt; DNA &lt;;213&gt;manmadesequence&lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 176 cacggtcaga acatggaatg cgaatgggac ccgtggacct gcgaacacat gttccgttac 60 &lt;210&gt; 177 &lt;211&gt; 60

&lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 177 ccgcgtctgc aggaagaatg cgtttgggac ccgtggacct gcgaacacat gccgctgcgt 60 &lt;210&gt;

&lt;211&gt; 60 &lt;212> DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA of a peptide capable of binding to Ang-2 &lt;400&gt; 178 cgtaccaccc aggaaaaatg cgaatgggac ccgtggacct gcgaacacat ggaatcccag 60 -291 - 1317281 丨 cage (287) &lt;Z10&gt; 17? &lt;2Ii&gt; 60 &lt;2I2&gt; DK?·· &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; % code DNA of peptide capable of binding to Ang-2

&lt;400&gt; P sgaccic twenty two aggaagact'g cgtttgggac ccgtggacct gcgaccacat ggtttcctcc 60 &lt;210&gt; I=: &lt;21i&gt; 5: &lt;212&gt;D&amp;&lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA of the peptide capable of binding to Ang-2 &lt;400&gt; 1Ξ3 csggr 223 gg gtcgtccgtg cgaatgggac ccgtggacct gcgaacacct ggaaggtctg 60 &lt;210&gt; 1=1 &lt;211&gt; 5:

&lt;212&gt; IKA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; • DNA of the peptide capable of binding to Ang-2 &lt;400&gt; :3 1 -aggcrrz=rc aggaagaatg cgcttgggac ccgtggacct gcgaccacat ggttggtctg 60 &lt;210&gt; 2 32 (288) (288) 60

1317281 &lt;211&gt; 60

&lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 182 ctgccgggtc aggaagactg cgaatgggac ccgtggacct gcgaacacat ggttcgttcc 60 &lt;210&gt; 183 &lt;211&gt; 60

&lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 183 ccgatgaacc aggttgaatg cgactgggac ccgtggacct gcgaacacat gccgcgttcc &lt;210&gt;

&lt;211&gt; 60 &lt;212&gt; DNA, &lt;213&gt; artificial sequence &lt;220&gt;&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 184 ttcggttggt ctcacggttg cgaatgggat ccgtggactt gcgaacacat gggttctacc 60 &lt ;210&gt; 185 &lt;211&gt; 60 •293 · 1317281 (289)

mMMMM

&lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 185 aaatccaccc aggacgactg cgactgggac ccgtggacct gcgaacacat ggttggtccg 60 &lt;210&gt; 186 &lt;2U&gt; 60 _

&lt;212> DNA &lt;213&gt; Artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 136 ggtccgcgta tctccacctg ccagtgggac ccgtggacct gcgaacacat ggaccagctg 60 &lt;210&gt; 187

&lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 187 tccaccatcg gtgacatgtg cgaatgggac ccgtggacct gcgctcacat gcaggttgac 60

&lt;210&gt; 188 &lt;211&gt; 60 &lt;212&gt; DNA -294·

60

60

1317281 (290) &lt;213&gt;man-made sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 188 gttctgggtg gtcagggttg cgaatgggac ccgtggacct gccgtctgct gcagggttgg &lt;210&gt; 189 &lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; artificial sequence&lt;220&gt;&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 189 gttctgggtg gtcagggttg ccagtgggac ccgtggacct gctcccacct ggaagacggt &lt;210&gt; 190 &lt;211&gt; 60

&lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 190 accaccatcg gttccatgtg cgaatgggac ccgtggacct gcgctcacat gcagggtggt 60 &lt;210&gt;191 &lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; 295- 1317281 (291) &lt;220&gt;

&lt;223&gt; DNA &lt;400&gt; 191 accaaaggta aatccgtttg ccagtgggac ccgtggacct gctcccacat gcagtccggt 60 &lt;210&gt; 192 encoding a peptide capable of binding to Ang-2

&lt;211&gt; 60 &lt;212&gt; DNA

&lt;213&gt;man-made sequence &lt;220&gt;

&lt;223&gt; DNA &lt;400&gt; 192 accaccatcg gttccatgtg ccagtgggac ccgtggacct gcgctcacat gcagggtggt 60 &lt;210&gt; 193 &lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; artificial sequence encoding peptide capable of binding to Ang-2

&lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 193 tgggttaacg aagttgtttg cgaatgggac ccgtggacct gcaaccactg ggacaccccg 60 &lt;210&gt; 194 &lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; -296· 1317281 (292) Face period fine &lt;220&gt;

&lt;223&gt; A DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 194 gttgttcagg ttggtatgtg 'ccagtgggac ccgtggacct gcaaacacat gcgtctgcag 60 &lt;210&gt; 195 &lt;211&gt; 60

&lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 195 gctgttggtt cccagacctg cgaatgggac ccgtggacct gcgctcacct ggttgaagtt 60 &lt;210&gt; 196

&lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; A DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 196 cagggtatga aaatgttctg cgaatgggac ccgtggacct gcgctcacat cgtttaccgt 60 &lt;210&gt; 197 &lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt;;220&gt; -297- 1317281 (293) &lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 197 accaccatcg gttccatgtg ccagtgggac ccgtggacct gcgaacacat gcagggtggt 60 &lt;210&gt; 198 &lt;211&gt; 60 &lt;212&gt;; DNA &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt;

DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 198 acctcccagc gtgttggttg cgaatgggac ccgtggacct gccagcacct gacctacacc 60 &lt;210&gt; 199 &lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; Artificial sequence &lt;220&gt;;223&gt; encoding can be combined with &lt;400&gt; 199 cagtggtcct ggccgccgtg cgaatgggac ccgtggacct gccagaccgt ttggccgtcc 60

&lt;210&gt; 200 &lt;211&gt; 60 &lt;212&gt; DNA

&lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 • 298 · 1317281 (294) &lt;400&gt; 200 ggtacctccc cgtccttctg ccagtgggac ccgtggacct gctcccacat ggttcagggt 60 &lt;210&gt;&lt;211&gt; 42 '

&lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 201 caggaagaat gcgaatggga cccatggact tgcgaacaca tg 42 &lt;210&gt; 202 &lt;211&gt; 66 &lt;212&gt; DNA &lt;213&gt; artificial sequence&lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 202 cagaactaca aaccgctgga cgaactggac gctaccctgt acgaacactt catcttccac 60 tacacc 66 &lt;210&gt;

&lt;211&gt; 66 &lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 -299 - 1317281 (295) mmm &lt;400&gt; 203 - ctgaacttca ccccgctgga cgaactggaa cagaccctgt acgaacagtg gaccctgcag 60 cagtcc 66 &lt;210&gt; 204 &lt;211&gt; 66 , &lt;;212&gt; DNA &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 • &lt;400&gt; 204 accaaattca acccgctgga cgaactggaa cagaccctgt acgaacagtg gaccctgcag 60 caccag 66 &lt;210&gt;&lt;211&gt; 66 &lt;212&gt; DNA &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; DN A encoding a peptide capable of binding to Ang-2 • &lt;400&gt; 205 gttaaattca aaccgctgga cgctctggaa cagaccctgt acgaacactg gatgttccag 60 Caggct 66 &lt;210&gt; 206 &lt;211&gt; 66

&lt;212&gt; DNA &lt;213&gt; artificial sequence -300-

1317281 (4) &lt;220&gt;

&lt;223&gt; DNA of a peptide capable of binding to Ang-2 &lt;400&gt; 206 gttaaataca aaccgctgga cgaactggac gaaatcctgt acgaacagca gaccttccag 60 gaacgt 66 &lt;210&gt; 207 &lt;211&gt;

&lt;212&gt; DNA &lt;213&gt; artificial sequence

&lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 207 accaacttca tgccgatgga cgacctggaa cagcgtctgt acgaacagtt catcctgcag 60 cagggt 66 &lt;210&gt; 208 &lt;211> 66 &lt;212&gt; DNA &lt;213&gt; sequence

&lt;220&gt;

&lt;223&gt; DNA of a peptide capable of binding to Ang-2 &lt;400&gt; 208 tccaaattca aaccgctgga cgaactggaa cagaccctgt acgaacagtg gaccctgcag 60 cacgct 66 &lt;210> 209

&lt;211&gt; 66 &lt;212&gt; DNA -301· 1317281 (297) &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 209 - cagaaattcc agccgctgga cgaactggaa cagaccctgt acgaacagtt catgctgcag 60 caggct 66

&lt;210&gt; 210 &lt;211&gt; 66 &lt;212&gt; DNA &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; DNA encoding peptide capable of binding to Ang-2 &lt;400&gt; 210 cagaacttca aaccgatgga cgaattggaa gacaccctgt Acaaacagtt cctgttccag 60 cactcc 66

&lt;210&gt; 211 &lt;211&gt; 66 &lt;212&gt; DNA &lt;213&gt; Artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 211 tacaaattca ccccgctgga cgacctggaa cagaccctgt acgaacagtg gaccctgcag 60 cacgtt 66 &lt;210&gt; 212 •302 · 1317281 (298) &lt;211&gt;65&lt;212&gt;DNA&lt;;213&gt; Artificial sequence_hetero_&lt;220&gt;&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2&lt;400&gt; 212 aggaatacga accgctggac gaactggacg aaaccctgta caaccagtgg atgttccacc agcgt&lt;210&gt;213&lt;211&gt; 66 &lt;212&gt DNA &lt;213&gt; artificial sequence &lt;220&gt;&lt;223&gt; DNA encoding peptide capable of binding to Ang-2 &lt;400&gt; 213 tccaacttca·tgccgctgga cgaactggaa cagaccctgt acgaacagtt catgctgcag caccag&lt;210&gt;214&lt;211&gt; 66 &lt;212&gt; DNA &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; DNA encoding peptide capable of binding to Ang-2&lt;400&gt; 214 cagaaatacc agccgctgga cgaactggac aaaaccctgt acgatcagtt catgctgcag cagggt 6065 60 66 60

-303· 66 1317281(299) &lt;210&gt; 215

&lt;211&gt; 66 &lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 215 cagaaattcc agccgctgga cgaactggaa gaaaccctgt acaaacagtg gaccctgcag 60 cagcgt 66 &lt;210&gt; 216 &lt;211&gt;

&lt;212&gt; DNA &lt;213&gt; Artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 216 gttaaataca aaccgctgga cgaactggac gaatggctgt accaccagtt caccctgcac 60 caccag 66 &lt;210&gt; 217 &lt;211&gt;

&lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 -304- 1317281 (300) &lt;400&gt; 217 cagaaattca tgccgctgga cgaactggac gaaatcctgt acgaacagtt catgttccag 60 cagtccc g7 &lt;210&gt;

&lt;211&gt; 66 1 &lt;212&gt; DNA

&lt;213&gt; artificial sequence &lt;220〉

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 218 cagaccttcc agccgctgga cgacctggaa gaatacttgt acgaacagtg gatccgtcgt 60 taccac 66 &lt;210&gt;

&lt;211&gt; 66 &lt;212&gt; DNA

&lt;213&gt;man-made sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 219 gaagactaca tgccgctgga cgctctggac gctcagctgt acgaacagtt catcctgctg 60 cacggt 66 &lt;210&gt; 220 &lt;211&gt;

&lt;212&gt; DNA &lt;213&gt; artificial sequence -305- 1317281 (301) — -ίτ—, I — ·Μ发明说^度页i &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 220 cacaccttcc agccgctgga cgaactggaa gaaaccctgt actaccagtg gctgtacgac 60 cagctg · 66 &lt;210&gt; 221 &lt;211&gt;

&lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 221 tacaaattca acccgatgga cgaactggaa cagaccctgt acgaagaatt cctgttccag 60 cacgct 66 &lt;210&gt; 222 &lt;211&gt;

&lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 222 accaactaca aaccgctgga cgaactggac gctaccctgt acgaacactg gatcctgcag 60 cactcc 66 &lt;210&gt;

&lt;211&gt; 66 &lt;212&gt; DNA -306 - 1317281 (302) mmmt &lt;213&gt; Artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 223 : 60 66 cagaaattca aaccgctgga cgaactggaa cagaccctgt acgaacagtg gaccctgcag cagcgt _ &lt;210&gt; 224 &lt;211&gt;

&lt;212&gt; DNA &lt;n3&gt; artificial sequence &lt;220&gt;

&lt;^3&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 224 60 66 accaaattcc agccgctgga cgaactggac cagaccctgt acgaacagtg gaccctgcag cagcgt &lt;210&gt; 225 &lt;211&gt; 66 &lt;212&gt; DNA &lt;213&gt; Sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400> 225 60 ββ accaacttcc agccgctgga cgaactggac cagaccctgt acgaacagtg gaccctgcag cagcgt &lt;210&gt; 226 -307· 1317281 (303) 二卢I,·&lt;211&gt; 60

&lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding &lt;400&gt; 226 aaattcaacc cgctggacga gctggaagag actctgtacg aacagtttac ttttcaacag 60 &lt;210&gt; 227 &lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; artificial sequence&lt;212&gt;;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 227 gctggtggta tgcgtccgta cgacggtatg ctgggttggc cgaactacga cgttcaggct 60

&lt;210&gt; 228 &lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; Artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 228 cagacttggg acgatccgtg catgcacatt ctgggtccgg ttacttggcg tcgttgcatc 60 &lt;210&gt; 229 &lt;211&gt; 60 1317281 _

&lt;212> DNA &lt;213&gt; Artificial sequence &lt;220&gt;

&lt;223&gt; A DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 229 gctccgggtc agcgtccgta cgacggtatg ctgggttggc cgacctacca gcgtatcgtt 60 &lt;210&gt; 230 &lt;211&gt; 60

&lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 230 tccggtcagc tgcgtccgtg cgaagaaatc ttcggttgcg gtacccagaa cctggctctg 60 &lt;210&gt;

&lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 231 ttcggtgaca aacgtccgct ggaatgcatg ttcggtggtc cgatccagct gtgcccgcgt 60

&lt;210&gt; 232 &lt;211&gt; 60 &lt;212&gt; DNA -309·

Ml: Meng Gu 1317281 (4) &lt;213&gt; Man-made sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 232 'ggtcaggacc tgcgtccgtg cgaagacatg ttcggttgcg gtaccaaaga ctggtacggt 60 &lt;210&gt; 233

&lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 233 ggtttcgaat actgcgacgg tatggaagac ccgttcacct tcggttgcga caaacagacc 60 &lt;210> 234

&lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a skin capable of binding to Ang-2 &lt;400&gt; 234 aaactggaat actgcgacgg tatggaagac ccgttcaccc agggttgcga caaccagtcc 60 &lt;210&gt;

&lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; Artificial sequence - 310 - 1317281 (306) Looking at &lt;220&gt;&lt;223&gt; DN A &lt;400&gt; encoding a peptide capable of binding to Ang-2 ; 235 ctgcaggaat ggtgcgaagg tgttgaagac ccgttcacct tcggttgcga aaaacagcgt 60 &lt;210&gt; 236 &lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;&lt;223&gt; coding DN of peptide capable of binding to Ang-2 A &lt;400&gt; 236 gctcaggact actgcgaagg tatggaagac ccgttcacct tcggttgcga aatgcagaaa

60 &lt;210&gt; 237 &lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;&lt;223&gt; DN A &lt;400&gt; 237 ctgctggact actgcgaagg encoding peptide capable of binding to Ang-2 Tgttcaggac ccgttcacct tcggttgcga aaacctggac

60 &lt;210&gt; 238 &lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; artificial sequence -311 - 1317281 (307)

.... , r. ' ·-» * '·«· ·-&amp;BMM &lt;220&gt;&lt;223&gt;DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 238 caccaggaat actgcgaagg tatggaagac ccgttcacct tcggttgcga Ataccagggt 60 &lt;210&gt; 239 &lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;'&lt;223&gt; DNA encoding peptide capable of binding to Ang-2 &lt;400&gt; 239 atgctggact Actgcgaagg tatggacgac ccgttcacct tcggttgcga caaacagatg

60 &lt;210> 240 &lt; 211 &gt; 60 &lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;&lt;223&gt; DNA encoding peptide capable of binding to Ang-2 &lt;400&gt; 240 ctgcaggact actgcgaagg tgttgaagac Ccgttcacct tcggttgcga aaaccagcgt

60 &lt;210&gt; 241 &lt;211&gt; 60 &lt;212&gt; DNA &lt;213&gt; Artificial sequence &lt;220&gt; •312-1317281 (308) Seven _ _ invention. Description, continued, 3⁄4

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 241 ctgcaggact actgcgaagg tgttgaagac ccgttcacct tcggttgcga aaaacagcgt 60 &lt;210&gt; 242 &lt;211&gt; 54 *

&lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 242 ttcgactact gcgaaggtgt tgaagacccg ttcactttcg gctgtgataa ccac &lt;210&gt; 243 &lt;211&gt; 250 &lt;212&gt; Protein &lt;213&gt; Artificial sequence&lt;220&gt;

&lt;223&gt; DNA encoding a peptide capable of binding to Ang-2 &lt;400&gt; 243

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 He 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 54

•313- (309) (309) 1317281 wmm

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 Gly Gly Gly Gly Gly Cys Thr Ala Gly Tyr His Trp 225 230 235 240

Asn Ser Asp Cys Glu Cys Cys Arg Arg Asn 245 250 &lt;210&gt; 244 &lt;211&gt; 29

&lt;212&gt; DNA &lt;213&gt; Artificial sequence 314· 1317281 (3.0) &lt;220&gt;&lt;2Z3&gt; Oligonucleotide &lt;400&gt; 244 caaacgaatg gatcctcatt aaagccaga &lt;210&gt; 245 &lt;211&gt; 42 &lt; 212> DNA &lt;213&gt; artificial sequence &lt;220&gt;&lt;223&gt; oligonucleic acid &lt;400&gt; 245 ggtggtgcgg ccgcactcga gactgttgaa agttgtttag ca &lt;210&gt; 246 &lt;211&gt; 29 &lt;212&gt; DNA &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt;oligonucleotide..&lt;400&gt; 246 caaacgaatg gatcctcatt aaagccaga &lt;210&gt; 247 &lt;211&gt; 43 &lt;212&gt;DNA&lt;213&gt; artificial sequence invention Continued fi 29

42

29 -315- Send her to see «iisssa 1317281 (311) &lt;220&gt;&lt;223> Oligonucleotide &lt;400&gt; 247 aacacaaaag tgcacagggt ggaggtggtg gtgcggccgc act 43 &lt;210&gt; 248 &lt;211&gt; 91 &lt;212&gt ; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Oligonucleotide &lt;400&gt;

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

Asn Lys Asn Asn Lys Asn Asn Lys Asn Asn Lys Asn Asn Lys Asn Asn 20 25 30

Lys Asn Asn Lys Ser Ala Arg Thr Gly Gly Gly Ala Thr Cys Cys Gly 35 40 45

Thr Gly Gly Ala Ser Cys Asn Asn Lys Asn Asn Lys Asn Asn Lys Asn 50 55 60

Asn Lys Asn Asn Lys Asn Asn Lys Asn Asn Lys Cys Ala Thr Thr Cys 65 70 75 80

Thr Cys Thr Cys Gly Ala Gly Ala Thr Cys Ala 85 90 &lt;210> 249 &lt;211&gt; 91 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence, 316 - Invention Description % 1317281 (312) &lt;220&gt;&lt;223&gt; Oligonucleotide &lt;400&gt; 249

Cys Ala Cys Ala'Gly Thr Gly Cys Ala Cys Ala Gly GXy Gly Thr Asn 15 10 15

Asn Lys Asn Asn Lys Asn Asn Lys Ala Ala Lys Cys Gly Lvs Cys Cys 20 25 30

Lys Asn Asn Lys Gly Ala Lys Gly Ala Lys Ala Thr Lys Thr Thr Lys 35 40 45

Gly Gly Lys Gly Gly Lys Asn Asn Lys Ala Cys Lys Thr Ala Lys Cys 50 55 60

Ala Lys Asn Asn Lys Asn Asn Lys Asn Asn Lys Cys Ala Thr Thr Cys 65 70 75 80

Thr Cys Thr Cys Gly Ala Gly Ala Thr Cys Ala 85 90 &lt;210&gt; 250 &lt;211&gt; 95

&lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Oligonucleotide &lt;400&gt; 250

Cys Ala Cys Ala Gly Thr Gly Cys Ala Cys Ma Gly Gly Gly Thr Asn 15 10 15

Asn Lys Ala Ala Lys Thr Thr Lys Ala Ala Lys Cys Cys Lys Cys Thr 20 25 30 317- Send-Description Continued: &quot;-s .- .. ' 1317281 (3〇)

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

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

Thr Lys Ala Cys Lys Thr Thr Lys Cys Ala Lys Cys Ala Lys Asn Asn 65 70 75 80

Lys Cys Ala Thr Thr Cys Thr Cys Thr Cys Gly Ala Gly Ala Thr 85 90 95 &lt;210&gt; 251

&lt;211&gt; 91 &lt;212&gt; Protein &lt; 213 &gt; artificial sequence &lt;220&gt;&lt;223&gt; Oligonucleotide &lt;400&gt;

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

Asn Lys Asn Asn Lys Asn Asn Lys Cys Ala Lys Gly Ala Lys Gly Ala 20 25 30

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

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

Thr Lys Asn Asn Lys Asn Asn Lys Asn Asn Lys Cys Ala Thr Thr Cys 65 70 75 80

Thr Cys Thr Cys Gly Ala Gly Ala Thr Cys Ala 85 90 318- (314) (314)1317281 劈 劈 确 « ** ** ** ** ** ** ** ** ϋ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ τ ·-| &lt;210&gt; 252 &lt;211&gt; 89 &lt;212&gt;Protein._ &lt;213&gt; Artificial sequence &lt;220&gt; - &lt;223&gt; Oligonucleotide &lt;400&gt;

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

Asn Lys Thr Thr Lys Gly Ala Lys Thr Ala Lys Asn Asn Lys Gly Ala 20 25 30

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

Thr Thr Lys' Ala Cys Lys Thr Thr Lys Gly Gly Lys Asn Asn Lys Gly 50 55 60

Ala Lys Ala- Ala Lys Cys Ala Lys Asn Asn Lys Cys Ala Thr Thr Cys 65 70 75 80

Thr Cys Thr Cys Gly Ala Gly Ala Thr 85 &lt;210&gt; 253 &lt;211&gt; 95 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Oligonucleotide &lt;400&gt; 253 1317281 (315) Yan Ming said that the monument continued in summer:

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

Asn Lys Ala Ala Lys Thr Thr Lys Ala Ala Lys Cys Cys Lys Cys Thr 20 25 30

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

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

Thr Lys Ala Cys Lys Thr Thr Lys Cys Ala Lys Cys Ala Lys Asn Asn 65 70 75 80

Lys Cys Ala Thr Thr Cys Thr Cys Thr Cys Gly Ala Gly Ala Thr 85 90 95 &lt;210&gt; 254 &lt;211&gt;

&lt;212&gt; DNA &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Oligonucleotide &lt;400&gt;254 cacagtgcac agggt &lt;210&gt; 255 &lt;211&gt; 16 &lt;212&gt; DNA &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Oligo: y: acid 15

320· 1317281 (316) Sound $ Description, &&lt;400&gt; 255 tgatctcgag agaatg &lt;210&gt; 256 &lt;211&gt; 21 &lt;212&gt; DNA 1 &lt;213&gt; Man-made sequence &lt;220&gt;&lt;223&gt; Oligonucleotide &lt;400&gt; 256 gttagctcac tcattaggca c &lt;210> 257 &lt;211&gt; 21 &lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;&lt;223&gt; Oligonucleotide &lt;400&gt; 257 gtaccgtaac actgagtttc g &lt;210&gt; 258 &lt;211&gt; 13

&lt;212&gt; DNA &lt;213&gt; artificial sequence &lt;220&gt;&lt;223&gt; Oligonucleus: a:acid &lt;400&gt; 258 ttacacttta tgcttccg 16

21 21

Further, the &lt;210&gt; Peptide &lt;220&gt;

&lt;221&gt; misc^ Feature &lt;222&gt; (2),.(2) &lt;223&gt; Xaa = Fc &lt;_&gt; 259

Met Xaa Gly Gly Gly Gly Gly Ala Gin Pro He Arg Gin Giu Glu Cys 15 l〇 15

Asp Trp Asp Pro Trp Thr Cys Glu His Met Trp Glu Val Leu Glu ' 20 25 30

&lt;210&gt; 260 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc_ feature &lt;;222&gt; (2)..(2) -322- 1317281 (318) Medullary surface &lt;223&gt; Xaa = Fc &lt;400&gt; 260

Met Xaa Gly Gly Gly Gly Gly Ala Gin Thr Asn lie Gin Glu Glu Cys 1 , 5 10 15

Glu Trp Asp Pro Trp Thr Cys Asp His Met Pro Gly Lys Leu Glu 20 25 30 &lt;210> 261 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Ang-2-bound peptide body &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (2).. (2) &lt;223&gt; Xaa * Fc &lt;400&gt;

Met Xaa Gly Gly Gly Gly Gly Ala Gin Trp Tyr Glu Gin Asp Ala Cys 15 10 15

Glu Trp Asp Pro Trp Thr Cys Glu His Met Ala Glu Val Leu Glu 20 &lt;210&gt; 262 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence 323 · Fen Ming Description Continued La 13172281 (4) &lt;220&gt &lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;221>misc_ feature &lt;222&gt; (2).. (2) &lt;223&gt; Xaa = Fc &lt;400&gt;

Met Xaa Gly Gly Gly Gly Gly Ala Gin Asn Arg Leu Gin Glu Val Cys 15 10 15

Glu Trp Asp Pro Trp Thr Cys Glu His Met Glu Asn Val Leu Glu 20 25 30 &lt;210&gt; 263 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Ang-2-bound peptide body &lt;220&gt;&lt;22i&gt; misc_feature &lt;222&gt; (2)..(2) &lt;223&gt; Xaa = Fc &lt;400&gt;

Met Xaa Gly Gly Gly Gly Gly Ala Gin Ala Ala Thr Gin Glu Glu Cys 1.5 10 15 -324- Extraordinary % Description Continued; 1317281 (320)

Glu Trp Asp Pro Trp Thr Cys Glu His Met Pro Arg Ser Leu Glu 20 25 30 &lt;210&gt; 264 &lt;211&gt; 31 &lt;212&gt; PRT &lt;213&gt; Protein artificial sequence &lt;220&gt;&lt;223&gt;&lt;;22〇&gt; Peptide capable of binding to Ang-2 &lt;221> &lt;222&gt; misc_ feature &lt;223&gt; Xaa = Fc &lt;400&gt;

Met Xaa Gly Gly Gly G丄y Gly Ala G丄n lieu Arg His Gin Glu Gly Cys 15 10 15

Glu Trp Asp Pro Trp Thr Cys Glu His Met Phe Asp Trp Leu Glu 20 25 30 &lt;210&gt; 265 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Ang-2-bound peptide body &lt;220&gt;&lt;221&gt; misc characteristic -325- (321) 1317281 &lt;222&gt; (2)..(2) &lt;223&gt; Xaa = Fc &lt;400&gt;

Met Xaa Gly Gly Gly Gly Gly Ala Gin Val Pro Arg Gin Lys Asp Cys 15 10 15

Glu Trp Asp Pro Trp Thr Cys Glu His Met Tyr Val Gly Leu Glu 20 25 30

&lt;210&gt; 266 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;221&gt;&lt;222&gt; (2) · · (2) &lt;223&gt; Xaa - Fc &lt;400&gt; 266

Met Xaa Gly Gly Gly Gly Gly Ala Gin Ser lie Ser His Glu Glu Cys 1 5 10 15

Glu Trp Asp Pro Trp Thr Cys Glu His Met Gin Val Gly Leu Glu 20 25 30 &lt;210&gt; 267 &lt;211&gt; 31 &lt;212&gt; Protein 326 - 1317281 (322) Fen Ming Description Continued: &lt;213&gt; Sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (2) . (2) &lt;223&gt; Xaa = Fc

&lt;400&gt; 267

Met Kaa Gly Gly Gly Gly Gly Ala Gin Trp Ala Ala Gin Glu Glu Cys 15 10 15

Glu Trp Asp Pro Trp Thr Cys Glu His Met Gly Arg Met Leu Glu 20 25 30 &lt;210&gt; 268 &lt;211&gt; 31 &lt;212&gt;Protein &lt;213&gt;

&lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt;misc-feature&lt;222&gt; (2)..(2) &lt;223&gt; Xaa = Fc &lt;400&gt ; 268

Met Xaa Gly Gly Gly Gly Gly Ala Gin Thr Trp Pro Gin Asp Lys Cys 15 10 15 -327- 1317281 (323)

Glu Trp Asp Pro Trp Thr Cys Glu His Met Gly Ser Thr Leu Glu 20 25 30 &lt;210&gt; 269 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;

&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc-features &lt;222&gt; (2)..(2) &lt;223&gt; Xaa = Fc &lt;400&gt;

Met Xaa Gly Gly Gly Gly Gly Ala Gin Gly His Ser Gin Glu Glu Cys 15 10 15

Gly Trp Asp Pro Trp Thr Cys Glu His Met Gly Thr Ser Leu Glu 20 25 · 30

&lt;210&gt; 270 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt; •328 - 1317281 (324) Will explain the Buddha page: &lt;22i&gt;misc_features&lt;222&gt; (2)..(2) &lt;223&gt; Xaa = Fc &lt;400&gt; 270

Met Xaa Gly Gly Gly Gly Gly Ala Gin Gin His Trp Gin Glu Glu Cys 15 10 15 25 30

Glu Trp Asp Pro Trp Thr Cys Asp His Met Pro Ser Lys Leu Glu 20

&lt;210&gt; 271 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt;&lt;222&gt; (2)..(2)

&lt;223&gt; Xaa = Fc &lt;400&gt; 271

Met Xaa Gly Gly Gly Gly Gly Ala Gin Asn Val Arg Gin Glu Lys Cys 15 10 15

Glu Trp Asp Pro Trp Thr Cys Glu His Met Pro Val Arg Leu Glu 20 25 30 &lt;210&gt; 272 &lt;211&gt; 31 329 - 奋衲娆明续瓦i

1317281 (325) &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc_features &lt;222&gt; )..(2) &lt;223&gt; Xaa = Fc &lt;400&gt; 272

Met Xaa Gly Gly Gly Gly Gly Ala Gin Lys Ser Gly Gin Val Glu Cys 15 10 15

Asn Trp Asp Pro Trp Thr Cys Glu His Met Pro Arg Asn Leu Glu 20 25 30 &lt;210&gt; 273 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Ang-2 bound peptide body &lt;220&gt;

&lt;22i&gt;misc_##L&lt;222> (2)..(2) &lt;223&gt; Xaa = Fc &lt;400&gt; 273

•330 - face 1317281 (326)

Met Xaa Gly Gly Gly Gly Gly Ala Gin Val Lys Thr G丄n Glu His Cys 15 10 15

Asp Trp Asp Pro Trp Thr Cys Glu His Met Arg Glu Trp Leu Glu 20 25 30 &lt;210&gt; 274 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence

&lt;220&gt;&lt;223&gt; A limb capable of binding to Ang-2 &lt;220&gt;&lt;221&gt; misc_feature &lt;222&gt; (2)..(2) &lt;223&gt; Xaa = Fc &lt;400&gt; 274

Met Xaa G丄y Gly Gly Gly Gly Ala Gin Ala Trp Gly Gin Glu Gly Cys 15 10 15

Asp Trp Asp Pro Trp Thr Cys Glu His Met Leu Pro Met Leu Glu 20 25 30 &lt;210&gt; 275 &lt;211&gt;&lt;212&gt;&lt;213&gt; 31 Protein artificial sequence &lt;220&gt;&lt;223&gt; Ang-2 combined belly-331 · 1317281 (327) &lt;220&gt;&lt;22i&gt; misc - feature &lt;222&gt; (2). (2) &lt;223&gt; Xaa = Fc &lt;400&gt;

Met Xaa Gly Gly Gly Gly Gly Ala Gin Pro Val Asn Gin Glu Asp Cys 15 10 15

Glu Trp Asp Pro Trp Thr Cys Glu His Met Pro Pro Met Leu Glu 20 25 30 &lt;210&gt; 276 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Ang-2-bound peptide body &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (2) .. (2) &lt;223&gt; Xaa = Fc &lt;400&gt;

Met Xaa Gly Gly Gly Gly Gly Ala Gin Arg Ala Pro Gin Glu Asp Cys 1 5 10 15

Glu Trp Asp Pro Trp Thr Cys Ala His Met Asp lie Lys Leu Glu 20 25 30 &lt;210&gt; 277 -332- 1317281 (328)

&lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc_features &lt;222&gt; 2)..(2.) &lt;223&gt; Xaa = Fc &lt;400&gt; 277

Met Xaa Gly Gly Gly Gly Gly Ala Gin His Gly Gin Asn Met Glu Cys 15 10 15

Glu Trp Asp Pro Trp Thr Cys Glu His Met Phe Arg Tyr Leu Glu 20 25 30 &lt;210&gt; 278 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Ang-2-bound peptide body &lt;220&gt;&lt;22i&gt; misc_characteristic &lt;222&gt; (2)..(2) &lt;223&gt; Xaa = Fc 333 - 1317281 (329) Description page title &lt;400&gt;; 278

Met Xaa Gly Gly Gly Gly GXy Als Gin Pro Arg Leu Gin Glu Glu Cys 1 5 10 15 Val Trp Asp Pro Trp Thr Cys Glu His Met Pro Leu Arg Leu Glu 20 25 30 &lt;210&gt; 279 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc - feature &lt;222&gt; (2).. (2) &lt;223&gt; Xaa = Fc &lt;400&gt; 279

Met Xaa Gly Gly Gly Gly Gly Ala Gin Arg Thr Thr Gin Glu Lys Cys 1 5 10 15

Glu Trp Asp Pro Trp Thr Cys Leu Glu 20 25 30 &lt;210&gt; (330) Description of the Invention Page Continuation &lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (2)..(2) &lt;223&gt; Xaa = Fc &lt ;400&gt; 280

Met Xaa Gly Gly Gly Gly Gly Ala Gin Gin Thr Ser Gin Glu Asp Cys 15 10 15

Val Trp Asp Pro Trp Thr Cys Asp His Met Val Ser Ser Leu Glu 20 25 30 &lt;210&gt; 281 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Ang-2 bound peptide body &lt;220&gt;

&lt;22i&gt; misc_ feature &lt;222&gt; (2) ., (2) &lt;223&gt; Xaa = Fc &lt;400&gt; 281

Met Xaa Gly Gly Gly Gly Gly Ala Gin Gin Val lie Gly Arg Pro Cys 15 10 15

Glu Trp Asp Pro Trp Thr Cys Glu His Leu Glu Gly Leu Leu Glu 20 25 3〇-335· 1317281 (331) Description of the Invention Continued &lt;210&gt; 282 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;

&lt;22i&gt; misc - feature &lt;222&gt; (2) ., (2) &lt;223&gt; Xaa = Fc &lt;400&gt; 282

Met Xaa Gly Gly Gly Gly Gly Ala Gin Trp Ala Gin Gin Glu Glu Cys 1 5 10 15

Ala Trp Asp Pro Trp Thr Cys Asp His Met Val Gly Leu Leu Glu 20 25 30 &lt;210&gt; 283 &lt;211&gt; 31 &lt;212&gt; Protein Lu &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Peptide bound to Ang-2 &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (2)..(2) &lt;223&gt; Xaa = Fc 336- 1317281 (332) Title page of the invention &lt;400&gt;; 283

Met Xaa Gly Gly Gly Gly Gly Ala Gin Leu Pro Gly Gin Glu Asp Cys 15 10 15

Glu Trp Asp Pro Trp Thr Cys Glu His Met Val Arg Ser Leu Glu 20 25 30 &lt;210&gt; 284 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Ang-2-bound peptide body &lt;220&gt;&lt;221&gt; misc_ feature &lt;222&gt; (2)..(2) &lt;223&gt; Xaa - Fc &lt;400&gt;

Met Xaa G丄y Gly Gly Gly Gly Ala Gin Pro Met Asn Gin Val Glu Cys 1 5 10 15

Asp Trp Asp Pro Trp Thr Cys Glu His Met Pro Arg Ser Leu Glu 20 25 30 &lt;210&gt; 285 &lt;211&gt; 31 &lt;212&gt;Protein&lt;213&gt; Artificial Sequence-337- 1317281 (333) Page &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;221&gt; misc_ feature &lt;222&gt; (2)..(2) &lt;223&gt; Xaa = Fc &lt;400&gt ; 285

Met Xaa Gly Gly Gly Gly Gly Ala Gin Phe Gly Trp Ser His Gly Cys 15 10 15

Glu Trp Asp Pro Trp Thr Cys Glu His Met Gly Ser Thr Leu Glu 20 25 30 &lt;210&gt; 286 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Ang-2-bound peptide body &lt;220&gt;&lt;22i&gt;misc_#·^:&lt;222&gt; (2)..(2) &lt;223&gt; Xaa - Fc &lt;400&gt;

Met Xaa G丄y Gly Gly Gly Gly Ala Gin Lys Ser Thr Gin Asp Asp Cys 15 10 15

Asp Τγό Asd Pro Trp Thr Cys Glu His Met Val Gly Pro Leu Glu ' 20 25 30 -338 - Invention Description Continued:

1317281 (334) &lt;21〇&gt; 287 &lt;211> 31 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;;22i&gt;misc-feature&lt;222&gt; (2)..(2) &lt;223&gt; Xaa = Fc &lt;400&gt; 287

Met Xaa Gly Gly Gly Gly Gly Ala Gin Gly Pro Arg lie Ser Thr Cys 15 10 15

Gin Trp Asp Pro Trp Thr Cys Glu His Met Asp Gin Leu Leu Glu 20 25 30 &lt;210&gt; 288 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Ang-2-bound peptide body &lt;220&gt;&lt;22i&gt; misc-feature

&lt;222&gt; (2). (2) -339- 1317281 _ (335) Description of the Invention Continued &lt;223&gt; Xaa « Fc &lt;400&gt; 288

Met Xaa Gly Gly Gly Gly Gly Ala Gin Ser Thr lie Gly Asp Met Cys 1 5 10 15

Glu Trp Asp Pro Trp Thr Cys Ala His Met Gin Val Asp Leu Glu 20 25 30 &lt;210&gt; 289 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Ang-2-bound peptide body &lt;220&gt;&lt;221&gt; misc_feature &lt;222&gt; (2)..(2) &lt;223&gt; Xaa = Fc &lt;400&gt;

Met Xaa Gly Gly Gly Gly Gly Ala Gin Val Leu Gly Gly Gin Gly Cys 15 10 15

Glu Trp Asp Pro Trp Thr Cys Arg Leu Leu Gin Gly Trp Leu Glu 20 25 30 &lt;210&gt; 290 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence - 340 - 1317281 (336) Page &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (2)..(2) &lt;223&gt; Xaa = Fc &lt;400&gt ; 290

Gly Cys 15 Glu

Met Xaa Gly Gly Gly Gly Gly Ala Gin Val Leu Gly Gly Gin 1 5 10

Gin Trp Asp Pro Trp Thr Cys Ser His Leu Glu Asp Gly Leu 20 25 30 &lt;210&gt; 291 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; -2 bound peptide body &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (2)..(2) &lt;223&gt; Xaa = Fc &lt;400&gt;

Met Cys 15

Met Xaa Gly Gly Gly Gly Gly Ala Gin Thr Thr lie Gly Ser 15 10 • 341 - 1317281 _ (337) Description of the Invention Continued

Glu Trp Asp Pro Trp Thr Cys Ala His Met Gin Gly Gly Leu Glu ' 20 25 30 &lt;210&gt; 292 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Peptide bound to Ang-2 &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (2)..(2) &lt;223&gt; Xaa = Fc &lt;400&gt;

Met Xaa Gly Gly Gly Gly Gly Ala Gin Thr Lys Gly Lys Ser Val Cys 15 10 15

Gin Trp Asp Pro Trp Thr Cys Ser His Met Gin Ser Gly Leu Glu 20 25 30 &lt;210> 293 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Ang-2-bound peptide body &lt;220&gt;&lt;22i&gt; misc characteristic -342 - 1317281 _ (338) Description of the invention continuation page &lt;222&gt; (2)..(2) &lt;223&gt; Xaa = Fc &lt;400&gt; 293

Met Xaa Gly Gly Gly Gly Gly Ala Gin Thr Thr lie Gly Ser Met Cys 1 5 10 15

Gin Trp Asp Pro Trp Thr Cys Ala His Met Gin Gly Gly Leu Glu 20 25 30 &lt;210&gt; 294 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Ang-2-bound peptide body &lt;220&gt;&lt;22i&gt;misc-feature&lt;222&gt; (2)..(2) &lt;223&gt; Xaa = Fc

&lt;400&gt; 294

Met Xaa Gly Gly Gly Gly Gly Ala Gin Trp Val Asn Glu Val Val Cys 1 5 10 15

Glu Trp Asp Pro Trp Thr Cys Asn His Trp Asp Thr Pro Leu Glu 20 25 30 &lt;210&gt; 295 &lt;211&gt; 31 &lt;212&gt; Protein 343 - 1317281 (339) Description of the Invention Continued &lt;213&gt;&lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (2) (2) &lt;223&gt; Xaa == Fc

&lt;400&gt; 295

Met Xaa Gly Gly Gly Gly Gly Ala Gin Val Val Gin Val Gly Met Cys 15 10 15

Gin Trp Asp Pro Trp Thr Cys Lys His Met Arg Leu Gin Leu Glu 20 25 30 &lt;210&gt; 296 &lt;211&gt; 31 &lt;212&gt;Protein &lt;213&gt;

&lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;221&gt; misc_ feature &lt;222&gt; (2)..(2) &lt;223&gt; = Fc &lt;400&gt;

Met Xaa Gly Gly Gly Gly Gly Ala Gin Ala Val Gly Ser Gin Thr Cys 15 l〇 -344- 1317281 (340) Description of the Invention Continued

Glu Trp Asp Pro Trp Thr Cys Ala His Leu Val Glu Val Leu Glu 20 25 30 &lt;210&gt; 297 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;

&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (2)..(2) &lt;223&gt; Xaa = Fc &lt;400> 297

Met Xaa Gly Gly Gly Gly Gly Ala Gin Gin Gly Met Lys Met Phe Cys 1 5 10 15

Glu Trp Asp Pro Trp Thr Cys Ala His lie Val Tyr Arg Leu Glu 20 25 · 30

&lt;210&gt; 298 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt; -345 (341) 341) 1317281 Description of the Invention Continued page &lt;22i&gt; misc_ Feature &lt;222&gt; (2)., (2) &lt;223> Xaa - Fc &lt;400&gt; 298 *

Met Xaa Gly Gly Gly Gly Gly Ala Gin Thr Thr lie Gly Ser Met Cys 15 10 15

Gin Trp Asp Pro Trp Thr Cys Giu His Met Gin Gly Gly Leu Glu 20 25 30 &lt;210&gt; 299 &lt;211&gt; 31 &lt;212&gt;Protein&lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Ang-2-bound peptide body &lt;220&gt;&lt;221&gt; misc_feature &lt;222&gt; (2) . (2) &lt;223&gt;X&amp;3 - Fc &lt;400&gt;

Met Xaa Gly Gly Gly Gly Gly Ala Gin Thr Ser Gin Arg Val Gly Cys 15 10 15

Glu Trp Asp Pro Trp Thr Cys Gin His Leu Thr Tyr Thr Leu Glu 20 25 30 &lt;210&gt; 300 &lt;211&gt; 31 346- Invention Description Continued

1317281 (342) &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc_features &lt;222&gt; )..(2) &lt;223&gt; Xaa = Fc &lt;400&gt; 300

Met Xaa Gly Gly Gly Gly Gly Ala Gin Gin Trp Ser Trp Pro Pro Cys 15 10 15

Glu Trp Asp Pro Trp Thr Cys Gin Thr Val Trp Pro Ser Leu Glu 20 25 30 &lt;210&gt; 301 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Ang-2-bound peptide body &lt;220&gt;&lt;221&gt;misc-feature&lt;222&gt; (2)..(2) &lt;223&gt; Xaa = Fc &lt;400&gt;

347- 1317281 Description of the Invention (343)

Met Xaa Gly Gly Gly Gly Gly Ala Gin Gly Thr Ser Pro Ser ?he Cys 1 5 10 15

Gin Trp Asp Pro Trp Thr Cys Ser His Met Val Gin Gly Leu Glu 20 25 30 &lt;210&gt; 302 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Ang-2-bound peptide body &lt;220&gt;&lt;22i&gt;misc_feature&lt;222&gt; (2)..(2) &lt;223&gt; Xaa = Fc &lt;400&gt; 302

Met Xaa Gly Gly Gly Gly Gly Ala Gin Thr Gin Gly Leu His Gin Cys 15 10 15

Glu Trp Asp Pro Trp Thr Cys Lys Val Leu Trp Pro Ser Leu Glu 20 ' 25 30 &lt;210&gt; 303 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Peptide bound to Ang-2 - 348 - 1317281 _ (344) I Description of the invention page &lt;220&gt;&lt;221&gt; misc_ feature &lt;222&gt; (2) · , (2) &lt;223&gt; Xaa = Fc &lt;400&gt; 303

Met Xaa Gly Gly Gly Gly Gly Ala Gin Val Trp Arg Ser Gin Val Cys 15 10 15

Gin Trp Asp Pro Trp Thr Cys Asn Leu Gly Gly Asp Trp Leu Glu 20 25 30 &lt;210&gt; 304 &lt;211&gt; 31

&lt;212&gt; PRT &lt;213&gt;

Protein artificial sequence &lt;220&gt;&lt;223&gt;&lt;220&gt; Peptide capable of binding to Ang-2 &lt;221&gt;

&lt;222&gt; misc_feature &lt;223&gt; Xaa = Fc &lt;400&gt; 304

Met Xaa Gly Gly Gly Gly Gly Ala Gin Asp Lys lie Leu Glu Glu Cys 1 5 10 15

Gin Trp Asp Pro Trp Thr Cys Gin Phe Phe Tyr Gly Ala Leu Glu 20 25 30 &lt;210&gt; 305 349· 1317281 _ (345) I Invention Description Continued &lt;211&gt; 31 &lt;212&gt;Protein&lt;213&gt; Sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;

&lt;22i&gt; misc - feature &lt;222&gt; (2). (2) &lt;223&gt; Xaa = Fc &lt;400&gt; 305

Met Xaa Gly Gly Gly Gly Gly Ala Gin Ala Thr Phe Ala Arg Gin Cys 1 5 10 15

Gin Trp Asp Pro Trp Thr Cys Ala Leu Gly Gly Asn Trp Leu Glu 20 25 30 &lt;210&gt; 306 &lt;211&gt; 31

&lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc^ feature &lt;222&gt; (2) . 2) &lt;223&gt; Xaa = Fc 350 - 1317281 (346) &lt;400&gt; 306

Met Xaa Gly Gly Gly Gly G丄y Ala Gin Giy Pro Ala Gin Glu Glu Cys 1 5 . * 10 15

Glu Trp Asp Pro Trp Thr Cys Glu Pro Leu Pro Leu Met Leu Glu 20 25 30 &lt;210&gt; 307 &lt;211&gt; 31 &lt;212&gt;Protein &lt;213&gt; Artificial sequence

&lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;221&gt; misc_ feature &lt;222&gt; (2).. (2) &lt;223&gt; Xaa = Fc &lt;400&gt; 307

Met Xaa Gly Gly Gly Gly Gly Ala Gin Arg Pro Glu Asp Met Cys Ser

Gin Trp Asp Pro Trp Thr Trp His Leu Gin Gly Tyr Cys Leu Glu 20 25 30 &lt;210&gt; 308 &lt;211&gt; 31 &lt;212&gt;Protein &lt;213&gt; Artificial Sequence -351 - &lt;220&gt; 1317281 (347 Inventive gas &lt;223&gt; Skin capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc_ special &lt;222&gt; (2)..(2) &lt;223&gt; Xaa =* Fc &lt;400&gt;; 308

Met Xaa Gly Gly Gly Gly Gly Ala Gin Leu Trp Gin Leu Ala Val Cys 15 10 15

Gin Trp Asp Pro Gin Thr Cys Asp His Met Gly Ala Leu Leu Glu 20 25 30 &lt;210&gt; 309 &lt;211&gt; 31 &lt;212&gt;Protein&lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Ang-2-bound peptibodies

j &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (2) .. (2) &lt;223&gt; Xaa = Fc &lt;400&gt; 309

Met Xaa Gly Gly Gly Gly Gly Ala Gin Thr Gin Leu Val Ser Leu Cys 1 5 l〇 15

Glu Trp Asp Pro Trp Thr Cys Arg Leu Leu Asp Gly Trp Leu Glu 20 25 30 •352 - 1317281 (348) &lt;210&gt; 310 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Man-made Sequence &lt;220&gt;;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;

&lt;22i&gt; misc - feature &lt;222&gt; (2) . . . (2) &lt;223&gt; Xsh - Fc &lt;400&gt; 310

Met Xaa Gly Gly Gly Gly Gly Ala Gin Met Gly Gly Ala Gly Arg Cys 15 10 15

Glu Trp Asp Pro Trp Thr Cys Gin Leu Leu Gin Gly Trp Leu G丄u • 20 25 30 &lt;210&gt; 311 &lt;211&gt; 31

&lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;221&gt; misc_feature &lt;222&gt; (2).. ( 2) &lt;223&gt; Xaa = Fc -353 - 1317281 (349) &lt;400> 311

Met Xaa Gly Gly Gly Gly Gly Ala Gin Met Phe Leu Pro Asn Glu Cys 1 5 10 15

Gin Trp Asp Pro Trp Thr Cys Ser Asn Leu Pro Glu Ala Leu Glu 20 25 30 &lt;210&gt; 312 &lt;211&gt; 31

&lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc - feature &lt;222&gt; (2).-( 2) &lt;223&gt; Xaa = Fc &lt;400&gt; 312

Met Xaa Gly Gly Gly Gly Gly Ala Gin Phe Gly Trp Ser His Gly Cys 15 10 15

Glu Trp Asp Pro Trp Thr Cys Arg Leu Leu Gin Gly Trp Leu Glu 20 25 30 &lt;210&gt; 313 &lt;211&gt; 31 &lt;212&gt;Protein &lt;213&gt; Artificial Sequence -354 - Invention Suspended Department 1317281 ( 350) &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc a feature &lt;222&gt; (2).. (2) &lt;223&gt; Xaa - Fc &lt;;400&gt; 313

Met Xaa Gly Gly Gly Gly Gly Ala Gin Trp Pro Gin Thr Glu Gly Cys 1 5 ICT 15

Gin Trp Asp Pro Trp Thr Cys Arg Leu Leu His Gly Trp Leu Glu 20 25 30 &lt;210&gt; 314 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;

&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc-characteristic &lt;222&gt; (2). (2) &lt;223&gt; Xaa = Fc &lt;400&gt;

Met Xaa Gly Gly Gly Gly Gly Ala Gin Pro Asp Thr Arg Gin Gly Cys 15 10 15

Gin Trp Asp Pro Trp Thr Cys Arg Leu Tyr Gly Met Trp Leu Glu 20 25 30 • 355 - Invention Continuation Page: 丨

1317281 (351) &lt;210&gt; 315 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;221&gt;;misc_Features&lt;222&gt; (2)..(2) &lt;223&gt; Xaa = Fc &lt;400&gt; 315

Met Xaa Gly Gly Gly Gly Gly Ala Gin Thr Trp Pro Gin Asp Lys Cys 15 10 15

Glu Trp Asp Pro Trp Thr Cys Arg Leu Leu Gin Gly Trp Leu Glu 20 25 30 &lt;210&gt; 316 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Ang-2-bound peptide body &lt;220&gt;&lt;22i&gt; misc-feature &lt;222&gt; (2)..(2)

-356 - 1317281 (352) Fen Lang explains poverty alleviation] &lt;223&gt; Xaa = Fc &lt;400&gt; 316

Met Xaa Gly Gly Gly Gly Gly Ala Gin Asp Lys lie Leu Glu Glu Cys 1,5 10 15

Glu Trp Asp Pro Trp Thr Cys Arg Leu Leu Gin Gly Trp Leu Glu 20 25 30 &lt;210&gt; 317 &lt;211&gt; 31

&lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;221&gt; misc_feature &lt;222&gt; (2) . ) &lt;223&gt; X3.3 — Fc &lt;400&gt; 317

Met Xaa Gly Gly Gly Gly Gly Ala Gin Ala Ala Thr Gin Glu Glu Cys 1 5 10 15 20 &lt;210&gt; 318 &lt;211&gt; 34 &lt;212&gt; Protein &lt;213&gt;

Glu Trp Asp Pro Trp Thr Cys Arg Leu Leu Gin Gly Trp Leu Glu 357 - 1317281 (353) I invented ^ &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; Misc_ Feature &lt;222&gt; (34)..(34) &lt;223&gt; Xaa = Fc &lt;400&gt; 318

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

Leu Tyr G丄u Gin Phe lie Leu Gin Gin Gly Leu Glu Gly Gly Gly Gly 20 25 30

Gly Xaa &lt;210&gt; 319 &lt;211&gt; 34 &lt;212&gt;protein &lt;213&gt; artificial sequence

&lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;221&gt; misc - feature &lt;222&gt; (34) .. (34) . &lt;223&gt; Xaa = Fc &lt;400&gt; 319 358 - Invention Description Continued 13172281 (354)

Met Gly Ala Gin Thr Asn Tyr Lys Pro Leu Asp Glu Leu Asp Ala Thr 15 10 15

Leu Tyr Glu His Trp lie Leu Gin His Ser Leu Glu Gly Gly Gly Gly 20 25 30

Gly Xaa &lt;210> 320 &lt;211> 34

&lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (34).. (34 ) &lt;223&gt; Xaa = Fc

&lt;400&gt; 320

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

Leu Tyr Asp Gin Phe Met Leu Gin Gin Gly Leu Glu Gly Gly Gly Gly 20 25 30

Gly Xaa &lt;210&gt; 321 &lt;211&gt; 34 &lt;212&gt; Protein 359· Illustrative Continuation Page 1 1317281 (355) &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; capable of binding to Ang-2 Peptide &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (34)..(34)

&lt;223&gt; Xaa = Fc &lt;400&gt; 321

Met Gly Ala Gin Leu Asn Phe Thr Pro Leu Asp Glu Leu Glu Gin Thr 15 10 15

Leu Tyr Glu Gin Trp Thr Leu Gin Gin Ser Leu Glu Gly Gly Gly Gly 20 25 30

Gly Xaa &lt;210&gt; 322 &lt;211&gt; 34

&lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc_feature &lt;222&gt; (34) . 34) &lt;223&gt; Xaa = Fc • 360-

Description of the Invention Continued page J 1317281 (356) &lt;400&gt; 322

Met Gly Ala Gin Gin Lys Phe Gin Pro Leu Asp Glu Leu Glu G丄n Thr 1 5 10 15

Leu Tyr Glu Gin Phe Met Leu Gin Gin Ala Leu Glu Gly Gly Gly Gly 20 25 30

Gly Xaa &lt;210&gt; 323

&lt;211&gt; 34 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;221&gt; misc_feature &lt;222&gt; 34)., (34) &lt;223&gt; Xaa = Fc

&lt;400&gt; 323

Met Gly Ala Gin Gin Glu Tyr Glu Pro Leu Asp Glu Leu Asp Glu Thr 15 10 15

Leu Tyr Asn Gin Trp Met Phe His Gin Arg Leu Glu Gly Gly Gly Gly 20 25 30

</ RTI> <RTIgt; Peptide &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (34)..(34)

&lt;223&gt; Xaa = Fc &lt;400&gt; 324

Met Gly Ala Gin Val Lys Tyr Lys Pro Leu Asp Glu Leu Asp Glu lie 15 10 15

Leu Tyr Glu Gin Gin Thr Phe Gin Glu Arg Leu Glu Gly Gly Gly Gly 20 25 30

Gly Xaa &lt;210&gt; 325

&lt;211&gt; 34 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc_feature &lt;222&gt; 34)..(34) &lt;223&gt; Xaa = Fc -362- Description of the Invention Continued. 1317281 (358) &lt;400&gt; 325

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

Leu Tyr Glu Gin Trp Thr Leu Gin Gin Arg Leu Glu Gly Gly Gly Gly 20 25 30

Gly Xaa &lt;210&gt; 326

&lt;211&gt; 34 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220> &lt;22i&gt; misc_ feature &lt;222&gt; )..(34) &lt;223&gt; Xaa = Fc

&lt;400&gt; 326

Met Gly Ala Gin Thr Asn Phe Gin Pro Leu Asp Glu Leu Asp Gin Thr 15 10 15

Leu Tyr Glu Gin Trp Thr Leu Gin Gin Arg Leu Glu Gly Gly Gly Gly 20 25 3CT

Gly Xaa &lt;210&gt; 327 363 - 1317281 (359) Description of the Invention Page Continuing Page] &lt;211&gt; 34 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; capable of binding to Ang-2 Peptide &lt;220&gt;&lt;22i&gt; misc_ feature

&lt;222&gt; (34)..(34) &lt;223&gt; Xaa = Fc &lt;220&gt;&lt;221&gt; Feature &lt;222> (34)..(34) &lt;223&gt; Xaa = Fc &lt;400&gt;; 327

Met Gly Ala Gin Gin Asn Phe Lys Pro Met Asp Glu Leu Glu Asp Thr 1 5 10 15

Leu Tyr Lys Gin Phe Leu Phe Gin His Ser Leu Glu Gly Gly Gly Gly 20 25 30

Gly Xaa &lt;210&gt; 328 &lt;211&gt; 34 &lt;212&gt;Protein &lt;213&gt; Artificial Sequence -364 - Fen Ming Enemy Continued; 1317281 (360) &lt;220&gt;&lt;223&gt; 2-bound peptide body &lt;220&gt;&lt;22i&gt; misc_ feature 4 &lt;222&gt; (34).. (34) &lt;223&gt; Xaa = Fc &lt;400&gt;

Met Gly Ala Gin Val Lys Tyr Lys Pro Leu Asp Glu Leu Asp Glu Trp 1 5 10 * 15

Leu Tyr His Gin Phe Thr Leu His His Gin Leu Glu Gly Gly Gly Gly 20 25 30

Gly Xaa &lt;210&gt; 329 &lt;211&gt; 34 &lt;212&gt;protein &lt;213&gt; artificial sequence

&lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (34)..(34) &lt;223&gt; Xaa = Fc &lt;400&gt; 329

Met Gly Ala Gin Tyr Lys Phe Thr Pro Leu Asp Asp Leu Glu Gin Thr 15 10 15 -365·

1317281 (361)

Leu Tyr Glu Gin Trp Thr Leu Gin His Val Leu Glu Gly Gly Gly Gly 20 . 25 30

Gly X£L3 &lt;210&gt; 330 &lt;211&gt; 34

&lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;221&gt; misc-feature &lt;222&gt; (34).. ( 34) &lt;223&gt; Xaa = Fc &lt;400&gt; 330

Met Gly Ala Gin Gin Asa Tyr Lys Pro Leu Asp Glu Leu Asp Ala Thr 1 5 10 15

Leu Tyr Glu His Phe lie Phe His Tyr Thr Leu Glu Gly Gly Gly Gly 20 25 30

Gly Xaa &lt;210&gt; 331 &lt;211&gt; 34 &lt;212&gt;Protein&lt;213&gt; Artificial sequence -366 - 1317281 _ (362) Explain that the title page &lt;220&gt;&lt;223&gt; can be combined with Ang-2 Peptide &lt;220&gt;&lt;221&gt; misc_ feature &lt;222&gt; (34)..(34) &lt;223&gt; Xaa - Fc &lt;400&gt;

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

Leu Tyr Glu His Trp Met Phe Gin Gin Ala Leu Glu Gly Gly Gly Gly 20 * 25 30

Gly Xaa &lt;210&gt; 332 &lt;211&gt; 34 &lt;212&gt;protein &lt;213&gt; artificial sequence

&lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc-characteristic &lt;222&gt; (34) (34) &lt;223&gt; Xaa = Fc &lt;400&gt;; 332 367 - 1317281 (363) Description of the Invention Continued

Met Gly Ala Gin Glu Asp Tyr Met Pro Leu Asd Ala Leu Asp Ala Gin 1 5 10 15

Leu Tyr Glu Gin Phe lie Leu Leu His Gly Leu Glu Gly Gly Gly Gly 20 25 30

Gly Xaa &lt;210&gt; 333 &lt;211&gt; 34 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt;_Features&lt;222&gt; (34)..(34) &lt;223&gt; Xaa = Fc &lt;400&gt; 333

Met Gly Ala Gin Tyr Lys Phe Asn Pro Met Asp Glu Leu Glu Gin Thr 1 5 10 15

Leu Tyr Glu Glu Phe Leu Phe Gin His Ala Leu Glu Gly Gly Gly Gly 20 25 30

Gly Xaa &lt;210&gt; 334 &lt; 211 &gt; 34 &lt;212&gt; Protein 368· Inventive Note Continued Page 1317281 (364) &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 Body &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (34)..(34) &lt;223&gt; Xaa = Fc

&lt;400&gt; 334 Met Gly Ala Gin Ser Asn Phe Met Pro Leu Asp Glu Leu Glu Gin Thr 1 5 10 15 Leu Tyr Glu Gin Phe Met Leu Gin His Gin Leu Glu Gly Gly Gly Gly 20 25 30

Gly Xaa &lt;210&gt; 335 &lt;211&gt; 34

&lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (34).. (34 } &lt;223&gt; Xaa - Fc -369 - Description of invention 碛^ 1317281 (365) &lt;400&gt; 335

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

Leu Tyr Lys Gin Trp Thr Leu Gin Gin Arg Leu Glu Gly Gly Gly Gly 20 25 30 G丄y Xaa &lt;210&gt; 336 &lt;211&gt; 34

&lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc_feature &lt;222&gt; (34).. ( 34) &lt;223&gt; Xaa = Fc

&lt;400&gt; 336

Met Gly Ala Gin Gin Lys Phe Met Pro Leu Asp Glu Leu Asp Glu lie 15 10 15

Leu Tyr Glu Gin Phe Met Phe Gin Gin Ser Leu Glu Gly Gly Gly Gly 20 25 30

Gly Xaa &lt;210&gt; 337 &lt;211&gt; 34 370- 1317281 _ (366) Description of the Invention Continued &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Compatible with Ang-2 Peptide &lt;220&gt;&lt;22i&gt; misc - feature &lt;222&gt; (34) .. (34)

&lt;223&gt; Xaa = Fc &lt;400&gt; 337

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

Leu Tyr Glu Gin Trp Thr Leu Gin His Gin Leu Glu Gly Gly Gly Gly 20 25 30

Gly Xaa &lt;210&gt; 338

&lt;211&gt; 34 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (34) &lt;223&gt; Xaa = Fc -371 - Description of the Invention Continued 13172281 (367) &lt;400&gt; 338

Met Gly Ala Gin His Thr Phe Gin Pro Leu Asp Glu Leu Glu Glu Thr 15 10 15

Leu Tyr Tyr Gin Trp Leu Tyr Asp Gin Leu Leu Glu Gly Gly Gly Gly 20 25 30

Gly Xaa &lt;210&gt; 339 &lt;211&gt; 34 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc_ Feature &lt;222&gt; (34) . . . (34) &lt;223&gt; Xaa = Fc &lt;400&gt; 339

Met Gly Ala Gin Gin Lys Phe Lys Pro Leu Asp Glu Leu Glu Gin Thr 1 5 10 15

Leu Tyr Glu Gin Trp Thr Leu Gin Gin Arg Leu Glu Gly Gly Gly Gly 20 25 30

Gly Xaa &lt;210&gt; 340 372· 1317281 _ (368) I invention description continuation page &lt;211&gt; 34 &lt;212&gt;protein &lt;213&gt; artificial sequence. column &lt;220&gt;&lt;223&gt; 2 bound peptibody &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (34) . . (34)

&lt;223&gt; Xaa = Fc &lt;400&gt; 340

Met Gly Ala Gin Gin Thr Phe Gin Pro Leu Asp Asp Leu Glu Glu Tyr 15 10 15

Leu Tyr Glu Gin Trp He Arg Arg Tyr His Leu Glu Gly Gly Gly Gly 20 25 30

Gly Xs.3 &lt;210&gt; 341

&lt;211&gt; 34 Protein artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc_feature &lt;222&gt; (34) . (34) - 373 - 1317281 _ (369) Description of the Invention Continued page <223> Xcid. = Fc &lt;400> 341

Met Gly Ala Gin Ser Lys Phe Lys Pro Leu Asp Glu Leu Glu Gin Thr 1 ; 5 10 15

Leu Tyr Glu Gin Trp Thr Leu Gin His Ala Leu Glu Gly Gly Gly Gly 20 25 30

Gly Xaa &lt;210&gt; 342 • &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt;&lt;220&gt; Peptide capable of binding to Ang-2 &lt;221&gt;Misc_Features&lt;222&gt; (2) ·. (2) &lt;223&gt; Xaa = Fc • &lt;400&gt; 342

Met Xaa Gly Gly Gly Gly Gly Ala Gin Ser Gly Gin Leu Axg Pro Cys 15 10 15

Glu Glu lie Phe Gly Cys Gly Thr Gin Asn Leu Ala Leu Leu Glu 20 25 30 &lt;210&gt; 343 &lt;211&gt; 31 374· 1317281 _ (370) I&quot; Invention Description Continued &lt;212&gt;Protein&lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt;misc-feature&lt;222&gt; (2). (2)

&lt;223&gt; Xaa = Fc &lt;400&gt; 343

Met Xaa Gly Gly Gly Gly Gly Ala Gin Ala Gly Gly Met Arg Pro Tyr 1 5 10 15

Asp Gly Met Leu Gly Trp Pro Asn Tyr Asp Val Gin Ala Leu Glu 20 25 30 &lt;210&gt; 344 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence

&lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (2) (2) &lt;223&gt; Xaa = Fc &lt;400&gt; 375 - 1317281 (371) Description of the Invention Continued

Met Xaa Gly Gly Gly Gly Gly Ala Gin Gly Gin Asp Leu Arg Pro Cys 1 5 10 15

Glu Asp Met Phe Gly Cys Gly Thr Lys Asp Trp Tyr Gly Leu Glu 20 25 30 &lt;210&gt; 345 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Ang-2-bound peptide body &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (2}.. (2) &lt;223&gt; Xaa = Fc &lt;400&gt;

Met Xaa Gly Gly Gly Gly Gly Ala Gin Ala Pro Gly Gin Arg Pro Tyr 1 5 10 15

Asp Gly Met Leu Gly Trp Pro Thr Tyr Gin Arg lie Val Leu Glu 20 &quot; 25 30 &lt;210&gt; 346 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Man-made Sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 - 376- (372) 13172281 Description of the continuation page &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (2)._ (2) . &lt;223&gt; Xaa = Fc &lt;400&gt; 346

Met Xaa Gly Gly Gly Gly Gly Ala Gin Gin Thr Trp Asp Asp Pro Cys 15 10 15

Met His lie Leu Gly Pro Val Thr Trp Arg Arg Cys lie Leu Glu 20 25

Gly Asp Lys Arg Pro Leu 15

&lt;210&gt; 347 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt;&lt;222&gt; (2)..(2) &lt;223&gt; Xaa = Fc &lt;400&gt; 347

Met Xaa Gly Gly Gly Gly Gly Ala Gin Ph 15 10

Glu Cys Met Phe Gly Gly Pro lie Gin Leu Cys Pro Arg Leu Glu 20 25 30 &lt;210&gt; 348 377-(373)1317281 Description of the Invention Continued &lt;211&gt; 25 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 in dogs&lt;220&gt;

&lt;22i&gt; misc_features &lt;222&gt; (2)..(2) &lt;223&gt; Xaa = Fc &lt;400&gt; 348

Met Xaa Gly Gly Gly Gly Gly Ala Gin Lys Arg Pro Cys Glu Glu lie 15 10 15

Phe Gly Gly Cys Thr Tyr Gin Leu Glu 20 25 &lt;210&gt; 349 &lt;211&gt; 31

&lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (2).. (2 &lt;223&gt; Xaa = Fc 378 - (374) 1317281 Description of the Invention Continued page &lt;400&gt; 349

Met Xaa Gly Gly Gly Gly Gly Ala Gin Leu Gin Glu Trp Cys Glu Gly 1 5 10 15

Val Glu Asp Pro ?he Thr Phe Gly Cys Glu Lys Gin Arg Leu Glu 20 25 30 &lt;210&gt; 350 &lt;211&gt; 31 &lt;212&gt;Protein &lt;213&gt; Artificial sequence

&lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (2)..(2) &lt;223&gt; Xaa - Fc &lt;400&gt; 350

Met Xaa Gly Gly Gly Gly Gly Ala Gin Met Leu Asp Tyr Cys Glu Gly 1 5 10 15

Met Asp Asp Pro Phe Thr Phe Gly Cys Asp Lys Gin Met Leu Glu 20 25 30 &lt;210&gt; 351 &lt;211&gt; Protein &lt;212&gt; Artificial Sequence &lt;213&gt; Artificial Sequence • 379- &lt;220&gt; 1317281 _ (375) I invention description continuation page &lt;223&gt; Peptibosome capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (2).. (2) &lt;223&gt; Χδ3 — Fc &lt;400&gt; 351

Met Xaa Gly Gly Gly Gly Gly Ala Gin His Gin Glu Tyr Cys Glu Gly 1 5 · 10 15

Met Glu Asp Pro Phe Thr Phe Gly Cys Glu Tyr Gin Gly Leu Glu 20 25 30 &lt;210&gt; 352 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Ang-2-bound peptide body &lt;220&gt;&lt;221&gt; misc_feature &lt;222&gt; (2) .. (2) &lt;223&gt; Xaa = Fc &lt;400&gt;

Met Xaa Gly G丄y Gly Gly Giy Ala Gin Leu Gin Asp Tyr Cys Glu Gly 1 5 10 15

Val Glu Asp Pro Phe Thr Phe Gly Cys Glu Asn Gin Arg Leu Glu 20 25 30-380- Invention Description Continued

</ RTI> <RTIgt; ;221&gt;misc-features&lt;222&gt; (2)·(2) &lt;223&gt; Xaa = Fc &lt;400&gt; 353

Met Xaa Gly Gly Gly Gly Gly Ala Gin Leu Leu Asp Tyr Cys Glu Gly 1 5 10 15

Val Gin Asp Pro Phe Thr Phe Gly Cys Glu Asn Leu Asp Leu Glu 20 &lt;210&gt; 354 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; 2-bound peptide body &lt;220&gt;&lt;22i&gt;misc-feature&lt;222&gt; (2).. (2) &lt;223&gt; Xaa = Fc

-381 - 1317281 (377) Description of invention continuation page &lt;400&gt; 354

Met Xaa Gly Gly Gly Gly Gly Ala Gin Gly Phe Glu Tyr Cys Asp Gly 1 5 · 10 15

Met Glu Asp Pro Phe Thr Phe Gly Cys Asp Lys Gin Thr Leu Glu 20 25 30 &lt;210> 355 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;&lt;223&gt; Ang-2-bound peptide body &lt;220&gt;&lt;22i&gt;misc-feature&lt;222&gt; (2) ·. (2) &lt;223&gt; Xaa = Fc &lt;400&gt; 355 '

Met Xaa Gly Gly Gly Gly Gly Ala Gin Ala Gin Asp Tyr Cys Glu Gly 15 10 15

Met Glu Asp Pro Phe Thr Phe Gly Cys Glu Met Gin Lys Leu Glu 20 25 30 &lt;210&gt; 356 &lt;211&gt; 31 &lt;212&gt;Protein&lt;213&gt; Artificial Sequence - 382 - 1317281 _ (378) I Invention Description Continued page &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc_feature « &lt;222&gt;- (2)..(2) &lt;223&gt; Xaa = Fc &lt;400&gt; 356

Met Xaa Gly Gly Gly Gly Gly Ala Gin Leu Gin Asp Tyr Cys Glu Gly 15 10 15

Val Glu Asp Pro Phe Thr Phe Gly Cys Glu Lys Gin Arg Leu Glu 20 25 30 &lt;210&gt; 357 &lt;211&gt; 31 &lt;212&gt; Protein &lt;213&gt; Artificial Sequence &lt;220&gt;

&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;22i&gt; misc_ feature &lt;222&gt; (2) . (2) &lt;223&gt; Xaa = Fc &lt;400&gt;

Met Xaa Gly Gly Gly Gly Gly Ala Gin Lys Leu Glu Tyr Cys Asp Gly 15 10 15

Met Glu Asp Pro Phe Thr Gin Glv Cys Asp Asn Gin Ser Leu Glu 20 25 30 -383 - 1317281 (379) Description of the Invention Continued &lt;210&gt; 358 &lt;211&gt; 29 &lt;212&gt; Protein &lt;213&gt; Sequence &lt;220&gt;&lt;223&gt; Peptide capable of binding to Ang-2 &lt;220&gt;&lt;221&gt; niisc_ feature &lt;222&gt; (2) . (2) &lt;223&gt; Xaa = Fc &lt;400&gt;; 353

Met Xaa Gly Gly Gly Gly Gly Ala Gin Phe Asp Tyr Cys Glu Gly Val 15 10 15

Glu Asp Pro Phe Thr Phe Gly Cys Asp Asn His Leu Glu 20 25 &lt;210&gt; 359 &lt;211&gt; 32 &lt;212&gt; Protein &lt;213&gt; Artificial sequence &lt;220&gt;&lt;223&gt; capable of with Ang-2 Combined peptide &lt;400&gt; 359

Cys Gly Gly Gly Gly Gly Ala Gin Thr Asn Phe Met Pro Met Asp Asp 15 10 15 Le Le Gin Gin Gly Leu Glu 30

Leu Glu Gin Arg Leu Tyr Glu Gin Phe 20 25

Claims (1)

13 17£&amp;ll23468? Tiger patent application Chinese patent application scope replacement (96 years 8 Patent Application 1. A polypeptide comprising at least one polypeptide selected from the group consisting of amino acid sequences consisting of sequence recognition: No. 4 and sequence recognition: No. 76 to sequence identification: No. 1 18, wherein The polypeptide is capable of binding to Ang-2 and its physiologically acceptable salts; peptide sequence identification number peptide sequence Con4-44 76 PIRQEHCDWDPWTCEHMWEV Con4-40 77 TNIQEECEWDPWTCDHMPGK Con4-4 78 WYEQDACEWDPWTCEHMAEV Con4-31 79 NRLQEVCEWDPWTCEHMENV Con4-C5 80 AATQEECEWDPWTCEHMPRS Con4-42 81 LRHQEGCEWDPWTCEHMFDW Con4-35 82 VPRQKDCEWDPWTCEHMYVG Con4-43 83 SISHEECEWDPWTCEHMQVG Con4-49 84 WAAQEECEWDPWTCEHMGRM Con4-27 85 TWPQDKCEWDPWTCEHMGST Con4-48 86 GHSQEECGWDPWTCEHMGTS Con4-46 87 QHWQEECEWDPWTCDHMPSK Con4-41 88 NVRQEKCEWDPWTCEHMPVR Con4-36 89 KSGQVECNWDPWTCEHMPRN Con4-34 90 VKTQEHCDWDPWTCEHMREW Con4-28 91 AWGQEGCDWDPWTCEHMLPM Con4-39 92 PVNQEDCEWDPWTCEHMPPM Con4-25 93 RAPQEDCEWDPWTCAHMDIK Con4-50 94 HGQNMECEWDPWTCEHMFRY Con4-38 95 PRLQEECVWDPWTCEHMPLR Con4-29 96 RTTQEKCEWDPWTCEHMESQ Con4 -47 97 QTSQEDCVWDPWTCDHMVSS Con4-20 98 QVIGRPCEWDPWTCEHLEGL Con4-45 99 WAQQEECAWDPWTCDHMVGL Con4-37 100 LPGQEDCEWDPWTCEHMVRS Con4-33 101 PMNQVECDWDPWTCEHMPRS 81211-960813.doc 1317281 AC2-Con4 102 FGWSHGCEWDPWTCEHMGST Con4-32 103 KSTQDDCDWDPWTCEHMVGP Con4-17 104 GPRISTCQWDPWTCEHMDQL Con4-8 105 STIGDMCEWDPWTCAHMQVD AC4-Con4 106 VLGGOGCEWDPWTCRLLOGW Con4-l 107 VLGGQGCQWDPWTCSHLEDG Con4-Cl 108 TTIGSMCEWDPWTCAHMQGG Con4-21 109 TKGKSVCQWDPWTCSHMQSG Con4-C2 110 TTIGSMCQWDPWTCAHMQGG Con4-18 111 WVNEVVCEWDPWTCNHWDTP Con4-19 112 WQVGMCQWDPWTCKHMRLO Con4-16 113 AVGSQTCEWDPWTCAHLVEV Con4-ll 114 QGMKMFCEWDPWTCAHIVYR Con4-C4 115 TTIGSMCQWDPWTCEHMQGG Con4-23 116 TSQRVGCEWDPWTCOHLTYT Con4-15 117 QWSWPPCEWDPWTCQTVWPS Con4-9 118 GTSPSFCQWDPWTCSHMVOG TN8-Con4 4 QEECEWDPWTCEHM 2. A fusion polypeptide comprising at least one The polypeptide and vehicle according to item 1 of the patent application, wherein the fusion polypeptide is capable of binding to Ang-2, and a physiologically acceptable salt thereof. 3. The fusion polypeptide according to claim 2, wherein the medium is at least one Fc functional site, polyethylene glycol, lipid, cholesterol group, carbohydrate and oligosaccharide. 4. The polypeptide according to claim 1 of the patent application, which is cyclic. 5. A dimer or multimer of a polypeptide according to item 1 of the patent application. 6. A composition having the following formula and its multimer: (X^^F^CX2),, 81211-960813.doc 1317281 where: F1 is the medium; X1 and X2 are independently selected from -(L^c-P1; -(Lbc-PhLSd-PlCLSe-P3; and -(L^e-P'-CL^d-P'-CL ^e-P'-CL^f-P4; wherein one or more of P1, P2, P3 and P4 independently comprise a group selected from the group consisting of sequence identification: number 4 and sequence identification: 76 to sequence identification: 118 The polypeptides; L1, L2, L3 and L4 are each independently a linker; and a, b, c, d, e and f are each independently 0 or 1, with the constraint that at least one of a and b is 1; And a physiologically acceptable salt thereof. 7. A composition according to claim 6 of the patent application having the formula: X^F1 or F, X2 and a physiologically acceptable salt thereof. The composition according to claim 6 of the patent application having the formula: F'-CL^e-P1 and a physiologically acceptable salt thereof. 9. The composition according to claim 6 of the scope of the patent application, It has the formula: F1-(L1)c-P1-(L2)d-P2 and its physiologically acceptable salt. 10. The composition according to claim 6 of the patent application having the following formula: 81211-960813.doc 1317281 P1-(L1)c-F1-(L2)d-P2 and its physiologically acceptable salts. 11. The composition according to claim 6 wherein F1 is an Fc functional moiety or a fragment thereof. 12. The composition according to claim 6 wherein F1 comprises the sequence recognition: amino acid sequence of number 60. 13. A polynucleotide encoding a polypeptide according to claim 1 of the patent application. 14. A performance carrier comprising a multi-nuclear acid according to item 13 of the scope of the patent application. 15. A host cell comprising a performance vector according to claim 14 of the scope of the patent application. 16. The host cell according to claim 15 wherein the cell is a prokaryotic cell. 17. The host cell according to claim 16 wherein the cell is an E. coli cell. 18. The host cell according to claim 15 wherein the cell is a eukaryotic cell. 19. A polypeptide according to sequence identification: No. 2 or a variant thereof. 20. A composition of matter having the formula and its multimer: (X^q-F'^X2), wherein: F1 is a medium; X1 and X2 are each independently selected from -(L^s-P1; - (L1)s-P1-(L2)t-P2; -(L1)sP,-(L2)t-P2-(L3)u-P3; ^ 81211-960813.doc 1317281 wherein one or more of P1, P2, P3 and P4 independently comprise a polypeptide selected from the group consisting of a sequence recognition: a polypeptide of number 2; L1, L2, L3 and L4 are each independently a linker; and q, r, s, t, u, and v are each independently 0 or 1, with the constraint that at least one of q and r is 1; and a physiologically acceptable salt thereof. 21. A pharmaceutical composition comprising an effective amount of a polypeptide according to item 1 of the patent application, and a pharmaceutically acceptable carrier. 22. Use of a fusion polypeptide according to item 2 of the patent scope for the preparation of a medicament for inhibiting unwanted angiogenesis in a mammal. 23. Use of a composition according to item 6 of the patent scope for the preparation of a medicament for inhibiting unwanted angiogenesis in a mammal. 24. Use of a fusion polypeptide according to item 2 of the patent scope for the manufacture of a medicament for the treatment of angiogenesis in an individual. 25. Use of a composition according to item 6 of the patent scope for the preparation of a medicament for the treatment of angiogenesis in an individual. 26. Use of a fusion polypeptide according to item 2 of the patent scope for the preparation of a medicament for modulating angiogenesis in a mammal. 27. Use of a composition according to item 6 of the patent scope for the preparation of a medicament for modulating angiogenesis in a mammal. 28. Use of a fusion polypeptide according to item 2 of the patent scope for the preparation of a medicament for inhibiting tumor growth characterized by unwanted angiogenesis in a mammal. 29. Use of a composition according to item 6 of the patent scope for the preparation of an angiogenic effect which is inhibited in mammals as being undesired 81211-960813.doc 1317281 The drug for tumor growth. 30. Use of a fusion polypeptide according to item 2 of the patent scope for the preparation of a medicament for use in the treatment of cancer with a chemotherapeutic agent in a mammal. 31. Use of a composition according to item 6 of the patent scope for the manufacture of a medicament for use in the treatment of cancer with a chemotherapeutic agent in a mammal. 32. The use according to claim 30 or 31, wherein the chemotherapeutic agent is at least one of 5-FU, CPT-1 1 and gram cancer. 33. Use of a fusion polypeptide according to item 2 of the patent scope for the preparation of a medicament for modulating at least one vascular permeability or plasma leakage in a mammal. 34. Use of a composition according to item 6 of the patent scope for the preparation of a medicament for modulating at least one vascular permeability or plasma leakage in a mammal. 35. Use of a fusion polypeptide according to item 2 of the patent scope for the preparation of a neovascular disease, obesity, hemangioblastoma, hemangioma, atherosclerosis, inflammation in at least one eye in a mammal A disease, an inflammatory condition, an atherosclerosis, endometriosis, a disease of a neoplasm, a disease associated with bones, or a drug for psoriasis. 36. Use of a composition according to item 6 of the patent scope for the preparation of a neovascular disease, obesity, hemangioblastoma, hemangioma, atherosclerosis, inflammation in at least one eye in a mammal A disease, an inflammatory condition, an atherosclerosis, endometriosis, a disease of a neoplasm, a disease associated with bones, or a drug for psoriasis. 81211-960813.doc