US 10,703,785 B2 · Targeting deregulated Wnt signaling in cancer using stabilized alpha-helices of BCL-9

Abstract

The invention provides structurally-constrained peptides by hydrocarbon stapling of a BCL9 HD2 helix for use as a therapeutic agent. The invention further provides methods and kits for use of the structurally-constrained peptide of the instant invention. The invention is based, at least in part, on the results provided herein demonstrating that hydrocarbon stapled helical peptides display excellent proteolytic, acid, and thermal stability, restore the native helical structure of the peptide, possess superior pharmacokinetic properties compared to the corresponding unmodified peptides, and are highly effective in binding to β-catenin in vitro, in cellulo, and in vivo, disrupting the BCL-9/β-catenin interaction, and thereby interfering with deregulated Wnt/β-catenin signaling for therapeutic benefit in a variety of human diseases including human cancer.

Claims20

Dependent claims

A structurally constrained peptide of an HD2 domain of BCL9 (BCL9-HD2), comprising at least one hydrocarbon staple, wherein the hydrocarbon staple is positioned so as to link a first BCL9-HD2 amino acid selected from the group consisting of Leu-351, Ser-352, Gln-353, Glu-354, Gln-355, His-358, Arg-359, Arg-361, Ser-362, Leu-363, Thr-365, Leu-366, Ile-369, Gln-370, Met-372, Leu-373, and Phe-374 with a second BCL9-HD2 amino acid selected from the group consisting of Leu-351, Ser-352, Gln-353, Glu-354, Gln-355, Leu-356, Glu-357, His-358, Arg-359, Glu-360, Arg-361, Ser-362, Leu-363, Thr-365, Leu-366, Arg-367, Asp-368, Ile-369, Gln-370, Arg-371, Met-372, Leu-373, and Phe-374.

A structurally constrained peptide of an HD2 domain of BCL9 (BCL9-HD2) comprising at least one hydrocarbon staple, wherein one hydrocarbon staple is located at the following positions within the BCL9 HD2 peptide:

i, i + 4 single staples:
XSQEXLEHRERSLQTLRDIQRBLF
(SEQ ID NO:
8)
LXQEQXEHRERSLQTLRDIQRBLF
(SEQ ID NO:
9)
LSXEQLXHRERSLQTLRDIQRBLF
(SEQ ID NO:
10)
LSQXQLEXRERSLQTLRDIQRBLF
(SEQ ID NO:
11)
LSQEXLEHXERSLQTLRDIQRBLF
(SEQ ID NO:
12)
LSQEQXEHRXRSLQTLRDIQRBLF
(SEQ ID NO:
13)
LSQEQLXHREXSLQTLRDIQRBLF
(SEQ ID NO:
14)
LSQEQLEXRERXLQTLRDIQRBLF
(SEQ ID NO:
15)
LSQEQLEHXERSXQTLRDIQRBLF
(SEQ ID NO:
16)
LSQEQLEHRXRSLXTLRDIQRBLF
(SEQ ID NO:
17)
LSQEQLEHREXSLQXLRDIQRBLF
(SEQ ID NO:
18)
LSQEQLEHRERXLQTXRDIQRBLF
(SEQ ID NO:
19)
LSQEQLEHRERSXQTLXDIQRBLF
(SEQ ID NO:
20)
LSQEQLEHRERSLXTLRXIQRBLF
(SEQ ID NO:
21)
LSQEQLEHRERSLQXLRDXQRBLF
(SEQ ID NO:
22)
LSQEQLEHRERSLQTXRDIXRBLF
(SEQ ID NO:
23)
LSQEQLEHRERSLQTLXDIQXBLF
(SEQ ID NO:
24)
LSQEQLEHRERSLQTLRXIQRXLF
(SEQ ID NO:
25)
LSQEQLEHRERSLQTLRDXQRBXF
(SEQ ID NO:
26)
LSQEQLEHRERSLQTLRDIXRBLX
(SEQ ID NO:
27)
i, i + 7 staples:
XSQEQLEXRERSLQTLRDIQRBLF
(SEQ ID NO:
28)
LXQEQLEHXERSLQTLRDIQRBLF
(SEQ ID NO:
29)
LSXEQLEHRXRSLQTLRDIQRBLF
(SEQ ID NO:
30)
LSQXQLEHREXSLQTLRDIQRBLF
(SEQ ID NO:
31)
LSQEXLEHRERXLQTLRDIQRBLF
(SEQ ID NO:
32)
LSQEQXEHRERSXQTLRDIQRBLF
(SEQ ID NO:
33)
LSQEQLXHRERSLXTLRDIQRBLF
(SEQ ID NO:
34)
LSQEQLEXRERSLQXLRDIQRBLF
(SEQ ID NO:
35)
LSQEQLEHXERSLQTXRDIQRBLF
(SEQ ID NO:
36)
LSQEQLEHRXRSLQTLXDIQRBLF
(SEQ ID NO:
37)
LSQEQLEHREXSLQTLRXIQRBLF
(SEQ ID NO:
38)
LSQEQLEHRERXLQTLRDXQRBLF
(SEQ ID NO:
39)
LSQEQLEHRERSXQTLRDIXRBLF
(SEQ ID NO:
40)
LSQEQLEHRERSLXTLRDIQXBLF
(SEQ ID NO:
41)
LSQEQLEHRERSLQXLRDIQRXLF
(SEQ ID NO:
42)
LSQEQLEHRERSLQTXRDIQRBXF
(SEQ ID NO:
43)
LSQEQLEHRERSLQTLXDIQRBLX
(SEQ ID NO:
44)
i, i + 3 single staples:
XSQXQLEHRERSLQTLRDIQRBLF
(SEQ ID NO:
45)
LXQEXLEHRERSLQTLRDIQRBLF
(SEQ ID NO:
46)
LSXEQXEHRERSLQTLRDIQRBLF
(SEQ ID NO:
47)
LSQEXLEXRERSLQTLRDIQRBLF
(SEQ ID NO:
48)
LSQEQXEHXERSLQTLRDIQRBLF
(SEQ ID NO:
49)
LSQEQLXHRXRSLQTLRDIQRBLF
(SEQ ID NO:
50)
LSQEQLEXREXSLQTLRDIQRBLF
(SEQ ID NO:
51)
LSQEQLEHXERXLQTLRDIQRBLF
(SEQ ID NO:
52)
LSQEQLEHRXRSXQTLRDIQRBLF
(SEQ ID NO:
53)
LSQEQLEHREXSLXTLRDIQRBLF
(SEQ ID NO:
54)
LSQEQLEHRERXLQXLRDIQRBLF
(SEQ ID NO:
55)
LSQEQLEHRERSXQTXRDIQRBLF
(SEQ ID NO:
56)
LSQEQLEHRERSLXTLXDIQRBLF
(SEQ ID NO:
57)
LSQEQLEHRERSLQXLRXIQRBLF
(SEQ ID NO:
58)
LSQEQLEHRERSLQTXRDXQRBLF
(SEQ ID NO:
59)
LSQEQLEHRERSLQTLXDIXRBLF
(SEQ ID NO:
60)
LSQEQLEHRERSLQTLRXIQXBLF
(SEQ ID NO:
61)
LSQEQLEHRERSLQTLRDXQRXLF
(SEQ ID NO:
62)
LSQEQLEHRERSLQTLRDIXRBXF
(SEQ ID NO:
63)
LSQEQLEHRERSLQTLRDIQXBLX
(SEQ ID NO:
64),
wherein X is any amino acid and further identifies the amino acid residues which are linked by a hydrocarbon staple.

172 more elements

A structurally constrained peptide of an HD2 domain of BCL9 (BCL9-HD2) comprising at least one hydrocarbon staple, wherein the at least one hydrocarbon staples is located at any of the following positions within the BCL9 HD9 domain:

Mixed i, i + 4;
i, i + 3;
and
i, i + 7 double staples:
XSQEXLEHRERSLQTLXDIQRBLX
(SEQ ID NO:
74)
XSQEXLEHRERSLQTXRDIQRBXF
(SEQ ID NO:
75)
XSQEXLEHRERSLQXLRDIQRXLF
(SEQ ID NO:
76)
XSQEXLEHRERSLQTLRDIQXBLX
(SEQ ID NO:
77)
XSQEXLEHRERSLQTLRDIXRBXF
(SEQ ID NO:
78)
XSQEXLEHRERSLQTLRDXQRXLF
(SEQ ID NO:
79)
XSQEQLEXRERSLQTLRDIXRBLX
(SEQ ID NO:
80)
XSQEQLEXRERSLQTLRDXQRBXF
(SEQ ID NO:
81)
XSQEQLEXRERSLQTLRXIQRXLF
(SEQ ID NO:
82)
XSQEQLEXRERSLQTLRDIQXBLX
(SEQ ID NO:
83)
XSQEQLEXRERSLQTLRDIXRBXF
(SEQ ID NO:
84)
XSQEQLEXRERSLQTLRDXQRXLF
(SEQ ID NO:
85)
XSQXQLEHRERSLQTLRDIXRBLX
(SEQ ID NO:
86)
XSQXQLEHRERSLQTLRDXQRBXF
(SEQ ID NO:
87)
XSQXQLEHRERSLQTLRXIQRXLF
(SEQ ID NO:
88)
XSQXQLEHRERSLQTLXDIQRBLX
(SEQ ID NO:
89)
XSQXQLEHRERSLQTXRDIQRBXF
(SEQ ID NO:
90)
XSQXQLEHRERSLQXLRDIQRXLF
(SEQ ID NO:
91)
Sequential i, i + 4 staples:
XSQEXLEHXERSLQTLRDIQRBLF
(SEQ ID NO:
92)
LXQEQXEHRXRSLQTLRDIQRBLF
(SEQ ID NO:
93)
LSXEQLXHREXSLQTLRDIQRBLF
(SEQ ID NO:
94)
LSQXQLEXRERXLQTLRDIQRBLF
(SEQ ID NO:
95)
LSQEXLEHXERSXQTLRDIQRBLF
(SEQ ID NO:
96)
LSQEQXEHRXRSLXTLRDIQRBLF
(SEQ ID NO:
97)
LSQEQLXHREXSLQXLRDIQRBLF
(SEQ ID NO:
98)
LSQEQLEXRERXLQTXRDIQRBLF
(SEQ ID NO:
99)
LSQEQLEHXERSXQTLXDIQRBLF
(SEQ ID NO:
100)
LSQEQLEHRXRSLXTLRXIQRBLF
(SEQ ID NO:
101)
LSQEQLEHREXSLQXLRDXQRBLF
(SEQ ID NO:
102)
LSQEQLEHRERXLQTXRDIXRBLF
(SEQ ID NO:
103)
LSQEQLEHRERSXQTLXDIQXBLF
(SEQ ID NO:
104)
LSQEQLEHRERSLXTLRXIQRXLF
(SEQ ID NO:
105)
LSQEQLEHRERSLQXLRDXQRBXF
(SEQ ID NO:
106)
LSQEQLEHRERSLQTXRDIXRBLX
(SEQ ID NO:
107)
Sequential i, i + 3 staples:
XSQXQLXHRERSLQTLRDIQRBLF
(SEQ ID NO:
108)
Sequential i, i + 7 staples:
NSQEQLEXRERSLQXLRDIQRBLF
(SEQ ID NO:
109)
Mixed sequential staples:
XSQXQLEXRERSLQTLRDIQRBLF
(SEQ ID NO:
110)
XSQXQLEHREXSLQTLRDIQRBLF
(SEQ ID NO:
111)
XSQEXLEXRERSLQTLRDIQRBLF
(SEQ ID NO:
112)
XSQEXLEHRERXLQTLRDIQRBLF
(SEQ ID NO:
113)
XSQEQLEXREXSLQTLRDIQRBLF
(SEQ ID NO:
114)
XSQEQLEXRERXLQTLRDIQRBLF.
(SEQ ID NO:
115).

131 more elements

Dependencies are inferred from the claim text.

Targeting deregulated Wnt signaling in cancer using stabilized alpha-helices of BCL-9

Life of this patent

Targeting deregulated Wnt signaling in cancer using stabilized alpha-helices of BCL-9

Life of this patent

Abstract

Claims20