Peptides binding to Bfl-1
AssigneeDana-Farber Cancer Institute, Inc.
InventorsLoren D. Walensky, Gregory H. Bird
Life of this patent
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Abstract
This disclosure features stapled peptide inhibitors (e.g., cysteine-reactive stapled peptides) of the anti-apoptotic protein, BFL-1, and methods of using same in the treatment of BFL-1 expressing cancers.
Claims76
A polypeptide that binds to Bfl-1, the polypeptide comprising:
- (a) the amino acid sequence set forth in SEQ ID NO:
- 22-36, 41-119, 184, 186, 195, 197, or 208, wherein the polypeptide comprises a non-natural amino acid bearing an electrophilic group or an electrophilic warhead at the N-terminus of the amino acid sequence;
- (b) an amino acid sequence that is at least 50% identical to the Bfl-1 interacting alpha-helical face of ATQLRRFGDKLNFRQ (SEQ ID NO:121) and that selectively binds Bfl-1 over MCL-1, wherein at least two amino acids in the amino acid sequence of SEQ ID NO:121 are substituted by non-natural amino acids with olefinic side chains, wherein F at position 7 of the amino acid sequence of SEQ ID NO:121 is substituted with an amino acid with a bulkier side chain, and wherein the polypeptide comprises a non-natural amino acid bearing an electrophilic group or an electrophilic warhead at the N-terminus of the amino acid sequence;
- (c) an amino acid sequence that is at least 50% identical to the Bfl-1 interacting alpha-helical face of an amino acid sequence set forth below:
- (i)
- (SEQ ID NO:
- 122)
- AELEVECATQLRRFGDKLNFRQKLLN;
- (ii)
- (SEQ ID NO:
- 123)
- EIWIAQELRRIGDEFNAYYARR;
- (iii)
- (SEQ ID NO:
- 124)
- DIIRNIARHLAQVGDSMDRSI;
- (iv)
- (SEQ ID NO:
- 125)
- SSTMGQVGRQLAIIGDDINRRY;
- (v)
- (SEQ ID NO:
- 126)
- QDASTKKLSESLKRIGDELDSNMEL;
- or
- (vi)
- (SEQ ID NO:
- 127)
- RLAEVCAVLLRLGDELEMIR,
- wherein the polypeptide selectively binds Bfl-1 over MCL-1;
- wherein at least two amino acids in each amino acid sequence are substituted by non-natural amino acids with olefinic side chains,
- wherein at least one cysteine if present in the polypeptide is optionally replaced by serine, and wherein at least one methionine if present in the polypeptide is optionally replaced by norleucine;
- and
- wherein the polypeptide comprises a non-natural amino acid bearing an electrophilic group or a non-amino acid warhead at the N-terminus of the amino acid sequence;
- (d) an amino acid sequence that is at least 50% identical to the Bfl-1 interacting alpha-helical face of JATQLRRFGDKLNFRQKLL (SEQ ID NO:
- 128) and that selectively binds Bfl-1 over MCL-1;
- wherein the polypeptide forms a covalent bond with Cys 55 of Bfl-1;
- wherein at least two amino acids in the amino acid sequence of SEQ ID NO:128 are substituted by non-natural amino acids with olefinic side chains, and wherein J is a non-natural amino acid bearing an electrophilic group, or an electrophilic warhead that does not comprise an amino acid;
- (e) an amino acid sequence that is at least 50% identical to the Bfl-1 interacting alpha-helical face of JEVESATQLRRFGDKLNFRQKLL (SEQ ID NO:129) and that selectively binds Bfl-1 over MCL-1;
- wherein the polypeptide forms a covalent bond with Cys 55 of Bfl-1;
- wherein at least two amino acids in the amino acid sequence of SEQ ID NO:129 are substituted by non-natural amino acids with olefinic side chains, and wherein J is a non-natural amino acid bearing an electrophilic group, or an electrophilic warhead that does not comprise an amino acid;
- (f) an amino acid sequence that is at least 50% identical to the Bfl-1 interacting alpha-helical face of JIAQELRRIGDEFNAYYARR (SEQ ID NO:
- 130) and that selectively binds Bfl-1 over MCL-1;
- wherein the polypeptide forms a covalent bond with Cys 55 of Bfl-1;
- wherein at least two amino acids in the amino acid sequence of SEQ ID NO:130 are substituted by non-natural amino acids with olefinic side chains, and wherein J is a non-natural amino acid bearing an electrophilic group, or an electrophilic warhead that does not comprise an amino acid;
- (g) an amino acid sequence with 1 to 5 amino acid substitutions relative to the amino acid sequence set forth in SEQ ID NO:
- 22-36, 41-119, 184, 186, 195, 197, or 208, wherein the polypeptide selectively binds Bfl-1 over MCL-1 and forms a covalent bond with Cys 55 of Bfl-1, and wherein the polypeptide comprises a non-natural amino acid bearings an electrophilic group or an electrophilic warhead at the N-terminus of the amino acid sequence;
- or
- (h) an amino acid sequence with 1 to 5 amino acid substitutions relative to the amino acid sequence set forth in SEQ ID NO:
- 128, wherein the polypeptide selectively binds Bfl-1 over MCL-1 and forms a covalent bond with Cys 55 of Bfl-1, and wherein the polypeptide comprises a non-natural amino acid bearing an electrophilic group or an electrophilic warhead at the N-terminus of the amino acid sequence.
A polypeptide that binds to Bfl-1, the polypeptide comprising an amino acid sequence that is at least 70% identical to the Bfl-1 interacting alpha-helical face of the amino acid sequence set forth in SEQ ID N:
- 22-36, 41-119, 184, 186, 195, 197, or 208, wherein the polypeptide comprises a non-natural amino acid bearing an electrophilic group or a non-amino acid warhead at the N-terminus in of the amino acid sequence.
A polypeptide that binds to Bfl-1, the polypeptide comprising the amino acid sequence JAT8LRRFGDXLNFRQ (SEQ ID NO:62) with 0 to 5 amino acid substitutions relative to the amino acid sequence of SEQ ID NO:62, wherein J is a non-natural electrophile containing amino acid or an electrophilic warhead that does not comprise an amino acid at the N-terminus of the amino acid sequence, 8 is R-octenyl alanine, and X is a non-natural amino acid with an olefinic side chain.
A polypeptide that binds to Bfl-1, the polypeptide comprising an amino acid sequence that is at least 70% identical to the Bfl-1 interacting alpha-helical face of the amino acid sequence set forth in SEQ ID NO:
- 131, wherein the polypeptide selectively binds Bfl-1 over MCL-1;
- wherein at least two amino acids in the amino acid sequence are substituted by non-natural amino acids with olefinic side chains, wherein the at least two amino acids are separated by three or six amino acids, and wherein the polypeptide comprises a non-natural amino acid bearing an electrophilic group at the N-terminus of the amino acid sequence.
Dependencies are inferred from the claim text.