Specific binding agents of human angiopoietin-2
Disclosed are peptides that bind to Ang-2. Also disclosed are peptibodies comprising the peptides, methods of making such peptides and peptibodies, and methods of treatment using such peptides and peptibodies.
1. A polypeptide capable of binding Ang-2 wherein said polypeptide comprises the amino acid sequence
WDPWT (SEQ ID NO: 65)
and wherein said polypeptide is from 5 to 50 amino acids in length, and physiologically acceptable salts thereof.
2. A polypeptide capable of binding Ang-2, wherein said polypeptide comprises the amino acid sequence
WDPWTC (SEQ ID NO: 66)
and physiologically acceptable salts thereof.
3. A polypeptide capable of binding Ang-2, wherein said polypeptide comprises the amino acid sequence
Cz 2 WDPWT (SEQ ID NO: 67)
wherein z 2 is an acidic or neutral polar amino acid residue, and physiologically acceptable salts thereof.
4. The polypeptide according to claim 3 , wherein said polypeptide further comprises the amino acid sequence
Cz 2 WDPWTC (SEQ ID NO: 68)
wherein z 2 is an acidic or neutral polar amino acid residue, and physiologically acceptable salts thereof.
5. A polypeptide capable of binding Ang-2 comprising an amino acid sequence of the formula:
a 1 a 2 a 3 Ca 5 WDPWTCa 12 a 13 a 14 (SEQ ID NO: 69)
wherein:
a 1 , a 2 , and a 3 are each independently amino acid residues;
a 5 is an amino acid residue;
a 12 is absent or an amino acid residue;
a 13 is absent or a neutral hydrophobic, neutral polar, or a basic amino acid residue;
a 14 is a neutral hydrophobic or neutral polar amino acid residue;
and physiologically acceptable salts thereof.
6. The polypeptide according to claim 5 wherein:
a 1 is V, I, P, W, G, S, Q, N, E, K, R, or H;
a 2 is V, P, M, G, S, Q, D, E, K, R, or H;
a 3 is A, V, P, M, F, T, G, D, E, K, or H;
a 8 is A, V, G, Q, N, D, or E;
a 12 is S, Q, N, D, E, K, or R;
a 13 is L, T, or H;
a 14 is V, L, I, W, or M;
and physiologically acceptable salts thereof.
7. The polypeptide according to claim 5 wherein:
a 1 is Q;
a 2 is E;
a 3 is E;
a 5 is D or E;
a 12 is D or E;
a 13 is H; and
a 14 is M;
and physiologically acceptable salts thereof.
8. A polypeptide capable of binding Ang-2 comprising an amino acid sequence of the formula:
b 1 b 2 b 3 b 4 b 5 b 6 Cb 8 WDPWTCb 15 b 16 b 17 b 18 b 19 b 20 (SEQ ID NO: 70)
wherein:
b 1 is absent or an amino acid residue;
b 2 is absent or a neutral hydrophobic, neutral polar, or a basic amino acid residue;
b 3 , b 4 , b 5 , and b 6 are each independently absent or amino acid residues;
b 8 is an amino acid residue;
b 15 is absent or an amino acid residue;
b 16 is absent or a neutral hydrophobic, neutral polar, or a basic amino acid residue;
b 17 is absent or a neutral hydrophobic or neutral polar amino acid residue;
b 18 , b 19 , and b 20 are each independently absent or amino acid residues;
and physiologically acceptable salts thereof.
9. The polypeptide according to claim 8 wherein:
b 1 is absent, or A, V, L, P, W, F, T, G, S, Q, N, K, R, or H;
b 2 is absent, or A, V, L, I, P, W, M, T, G, S, Y, N, K, R, or H;
b 3 is absent, or A, L, I, P, W, M, T, G, S, Q, N, E, R, or H;
b 4 is V, I, P, W, G, S, Q, N, E, K, R, or H;
b 5 is V, P, M, G, S, Q, D, E, K, R, or H;
b 6 is A, V, P, M, F, T, G, D, E, K, or H;
b 8 is A, V, G, Q, N, D, or E;
b 15 is S, Q, N, D, E, K, or R;
b 16 is L, T, or H;
b 17 is V, L, I, W, or M;
b 18 is absent, or A, V, L, P, W, F, T, G, Y, Q, D, E, or R;
b 19 is absent, or V, L, I, P, T, G, S, Y, Q, N, D, E, or R; and
b 20 is absent, or V, L, P, W, M, T, G, S, Y, Q, N, D, K, or R;
and physiologically acceptable salts thereof.
10. The polypeptide according to claim 8 wherein:
b 1 is absent, or P, or T;
b 2 is absent, or I, or N;
b 3 is absent, or R, or I;
b 4 is Q;
b 5 is E;
b 6 is E;
b 8 is D or E;
b 15 is D or E;
b 16 is H;
b 17 is M;
b 18 is absent, or W, or P;
b 19 is absent, or G, or E; and
b 20 is absent, or V, or K,
and physiologically acceptable salts thereof.
11. A polypeptide comprising at least one amino acid sequence selected from the group consisting of SEQ ID NO: 4, and SEQ ID NO: 76 to SEQ ID NO: 118, inclusive, wherein said polypeptide is capable of binding to Ang-2, and physiologically acceptable salts thereof;
PEPTIDE
SEQ ID NO.
PEPTIDE SEQUENCE
Con4-44
76
PIRQEECDWDPWTCEHMWEV
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
AC2-Con4
102
FGWSHGCEWDPWTCEHMGST
Con4-32
103
KSTQDDCDWDPWTCEHMVGP
Con4-17
104
GPRISTCQWDPWTCEHMDQL
Con4-8
105
STIGDMCEWDPWTCAHMQVD
AC4-Con4
106
VLGGQGCEWDPWTCRLLQGW
Con4-1
107
VLGGQGCQWDPWTCSHLEDG
Con4-C1
108
TTIGSMCEWDPWTCAHMQGG
Con4-21
109
TKGKSVCQWDPWTCSHMQSG
Con4-C2
110
TTIGSMCQWDPWTCAHMQGG
Con4-18
111
WVNEVVCEWDPWTCNHWDTP
Con4-19
112
VVQVGMCQWDPWTCKHMRLQ
Con4-16
113
AVGSQTCEWDPWTCAHLVEV
Con4-11
114
QGMKMFCEWDPWTCAHIVYR
Con4-C4
115
TTIGSMCQWDPWTCEHMQGG
Con4-23
116
TSQRVGCEWDPWTCQHLTYT
Con4-15
117
QWSWPPCEWDPWTCQTVWPS
Con4-9
118
GTSPSFCQWDPWTCSHMVQG
TN8-Con4
4
QEECEWDPWTCEHM.
12. A fusion polypeptide comprising at least one peptide according to claims 1 , 5 , 8 , or 11 and a vehicle, wherein said fusion polypeptide is capable of binding to Ang-2, and physiologically acceptable salts thereof.
13. The fusion polypeptide according to claim 12 wherein said vehicle is at least one of an Fc domain, polyethylene glycol, a lipid, a cholesterol group, a carbohydrate, and an oligosaccharide.
14. The polypeptide according to claims 1 , 5 , 8 , or 11 which is cyclic.
15. A dimer or multimer of the polypeptides according to claims 1 , 5 , 8 , or 11 .
16. A composition of matter having the formula:
(X 1 ) a -F 1 -(X 2 ) b
and multimers thereof, wherein:
F 1 is a vehicle;
X 1 and X 2 are each independently selected from
-(L 1 ) c -P 1 ;
-(L 1 ) c -P 1 -(L 2 ) d -P 2 ;
-(L 1 ) c -P 1 -(L 2 ) d -P 2 -(L 3 ) e -P 3 ; and
-(L 1 ) c -P 1 -(L 2 ) d -P 2 -(L 3 ) e -P 3 -(L 4 ) f -P 4 ;
wherein one or more of P 1 , P 2 , P 3 , and P 4 each independently comprise a polypeptide selected from the group consisting of:
(a) the amino acid sequence WDPWT (SEQ ID NO: 65), wherein said polypeptide is from 5 to 50 amino acids in length;
(b) the amino acid sequence WDPWTC (SEQ ID NO: 66),
(c) the amino acid sequence Cz 2 WDPWT (SEQ ID NO: 67), and
(d) the amino acid sequence Cz 2 WDPWTC (SEQ ID NO: 68),
wherein z 2 is an acidic or neutral polar amino acid residue;
L 1 , L 2 , L 3 , and L 4 are each independently linkers; and
a, b, c, d, e, and f are each independently 0 or 1, provided that at least one of a and b is 1;
and physiologically acceptable salts thereof.
17. The composition of matter according to claim 16 wherein z 2 is A, V, G, Q, N, D, or E.
18. The composition of matter according to claim 16 wherein one or more of P 1 , P 2 , P 3 , and P 4 each independently comprise a polypeptide selected from the group consisting of SEQ ID NO: 4, and SEQ ID NO: 76 to SEQ ID NO: 118 inclusive.
19. The composition of matter of claim 16 of the formulae:
X 1 -F 1
or
F 1 -X 2
and physiologically acceptable salts thereof.
20. The composition of matter of claim 16 of the formula:
F 1 -(L 1 ) c -P 1
and physiologically acceptable salts thereof.
21. The composition of matter of claim 16 of the formula:
F 1 -(L 1 ) c -P 1 -(L 2 ) d -P 2
and physiologically acceptable salts thereof.
22. The composition of matter of claim 16 of the formula:
P 1 -(L 1 ) c -F 1 -(L 2 ) d -P 2
and physiologically acceptable salts thereof.
23. The composition of matter of claim 16 , wherein F 1 is an Fc domain or fragment thereof.
24. The composition of matter of claim 16 wherein F 1 comprises the amino acid sequence of SEQ ID NO: 60.
25. A pharmaceutical composition comprising an effective amount of a polypeptide according to claim 1 in admixture with a pharmaceutically acceptable carrier thereof.