Stabilized alpha helical peptides and uses thereof
AssigneesDana-Farber Cancer Institute, Inc., President and Fellows of Harvard College
InventorsLoren D. Walensky, Stanley J. Korsmeyer, Gregory L. Verdine
Life of this patent
Estimating the term…
Abstract
Novel polypeptides and methods of making and using the same are described herein. The polypeptides include cross-linking (“hydrocarbon stapling”) moieties to provide a tether between two amino acid moieties, which constrains the secondary structure of the polypeptide. The polypeptides described herein can be used to treat diseases characterized by excessive or inadequate cellular death.
Claims30
A method of producing a modified cell-permeable α-helical polypeptide, the method comprising:
- (a) selecting a precursor polypeptide comprising at least two reactive residues;
- and
- (b) reacting the precursor polypeptide in conditions sufficient to promote formation of a crosslink spanning at least one turn of an alpha helix between two of the at least two reactive residues, thereby forming the modified cell-permeable α-helical polypeptide;
- wherein the modified cell-permeable α-helical polypeptide is cell-permeable as determined by a cell permeability study;
- and
- wherein at least one of the at least two reactive residues is an α,α-disubstituted amino acid.
A method of producing a modified cell-permeable α-helical polypeptide, the method comprising:
- (a) selecting a precursor polypeptide comprising at least two reactive residues, each of the at least two reactive residues comprising a terminal olefin group;
- and
- (b) reacting the precursor polypeptide in conditions sufficient to promote formation of a crosslink between the terminal olefin groups of two of the at least two reactive residues, thereby forming the modified cell-permeable α-helical polypeptide, wherein the crosslink spans at least one turn of an alpha helix;
- wherein the modified cell-permeable α-helical polypeptide is cell-permeable as determined by a cell permeability study.
A method of producing a modified cell-permeable α-helical polypeptide, the method comprising
- (a) selecting a precursor polypeptide comprising at least two reactive residues;
- and
- (b) reacting the precursor polypeptide in conditions sufficient to promote formation of a crosslink between two of the at least two reactive residues, thereby forming the modified cell-permeable α-helical polypeptide;
- wherein the modified cell-permeable α-helical polypeptide is cell-permeable as determined by a cell permeability study;
- and
- wherein the modified cell-permeable α-helical polypeptide has the formula:
- wherein:
- each R1 and R2 are independently H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkylalkyl, heteroarylalkyl, or heterocyclylalkyl;
- R3 is alkyl, alkenyl, alkynyl, or [R4—K—R4]n;
- each of which is substituted with 0-6 R5;
- R4 is alkyl, alkenyl, or alkynyl;
- R5 is halo, alkyl, OR6, N(R6)2, SR6, SOR6, SO2R6, CO2R6, R6, a fluorescent moiety, or a radioisotope;
- K is O, S, SO, SO2, CO, CO2, CONR6, or
- R6 is H, alkyl, or a therapeutic agent;
- n is an integer from 1-4;
- x is an integer from 2-10;
- each y is independently an integer from 1-100;
- z is an integer from 1-10;
- and
- each Xaa is independently an amino acid.
A method of producing a pharmaceutical composition comprising a modified cell-permeable α-helical polypeptide, the method comprising:
- (a) selecting a precursor polypeptide comprising at least two reactive residues;
- (b) reacting the precursor polypeptide in conditions sufficient to promote formation of a crosslink spanning at least one turn of an alpha helix between two of the at least two reactive residues, thereby forming the modified cell-permeable α-helical polypeptide;
- wherein the modified cell-permeable α-helical polypeptide is cell-permeable as determined by a cell permeability study;
- wherein at least one of the at least two reactive residues is an α,α-disubstituted amino acid;
- and
- (c) combining the modified cell-permeable α-helical polypeptide with a pharmaceutically acceptable carrier or adjuvant.
A method of producing a pharmaceutical composition comprising a modified cell-permeable α-helical polypeptide, the method comprising:
- (a) selecting a precursor polypeptide comprising at least two reactive residues, each of the at least two reactive residues comprising a terminal olefin group;
- (b) reacting the precursor polypeptide in conditions sufficient to promote formation of a crosslink between the terminal olefin groups of two of the at least two reactive residues, thereby forming the modified cell-permeable α-helical polypeptide;
- wherein the crosslink spans at least one turn of an alpha helix;
- wherein the modified cell-permeable α-helical polypeptide is cell-permeable as determined by a cell permeability study;
- and
- (c) combining the modified cell-permeable α-helical polypeptide with a pharmaceutically acceptable carrier or adjuvant.
A method of producing a pharmaceutical composition comprising a modified cell-permeable α-helical polypeptide, the method comprising
- (a) selecting a precursor polypeptide comprising at least two reactive residues;
- (b) reacting the precursor polypeptide in conditions sufficient to promote formation of a crosslink between two of the at least two reactive residues, thereby forming the modified cell-permeable α-helical polypeptide;
- wherein the modified cell-permeable α-helical polypeptide is cell-permeable as determined by a cell permeability study;
- and
- wherein the modified cell-permeable α-helical polypeptide has the formula:
- wherein:
- each R1 and R2 are independently H, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkylalkyl, heteroarylalkyl, or heterocyclylalkyl;
- R3 is alkyl, alkenyl, alkynyl, or [R4—K—R4]n;
- each of which is substituted with 0-6 R5;
- R4 is alkyl, alkenyl, or alkynyl;
- R5 is halo, alkyl, OR6, N(R6)2, SR6, SOR6, SO2R6, CO2R6, R6, a fluorescent moiety, or a radioisotope;
- K is O, S, SO, SO2, CO, CO2, CONR6, or
- R6 is H, alkyl, or a therapeutic agent;
- n is an integer from 1-4;
- x is an integer from 2-10;
- each y is independently an integer from 1-100;
- z is an integer from 1-10;
- and
- each Xaa is independently an amino acid;
- and
- (c) combining the modified cell-permeable α-helical polypeptide with a pharmaceutically acceptable carrier or adjuvant.
Dependencies are inferred from the claim text.