Thiol-yne based peptide stapling and uses thereof

A compound having a structure represented by a formula:

• wherein each occurrence of is an optional covalent bond, thereby signifying a single bond or a double bond;
• wherein each of x and z is independently an integer having a value of 0 to 100;
• wherein y is an integer having a value of 2 to 10;
• wherein each instance of Xaa in [Xaa]x is, independently, a natural or unnatural amino acid residue when x is 1 to 100;
• and wherein [Xaa]x is hydrogen when x is 0;
• wherein each instance of Xaa in [Xaa]y is, independently, a natural or unnatural amino acid;
• wherein each instance of Xaa in [Xaa]z is, independently, a natural or unnatural amino acid when z is 1 to 100;
• and wherein [Xaa]z is hydroxy when z is 0;
• wherein each of R1a and R1b is independently hydrogen or an amino protecting group;
• wherein each of R2a and R2b is independently hydrogen or C1-C4 alkyl;
• wherein each of R3a and R3b is independently a C1-C4 alkylene;
• wherein Z is C5-C12 alkylene, —(C2-C6 alkylene)-O—(C2-C6 alkylene)-, —(CH2)3—(OCH2CH2)q—O—(CH2)3—, substituted with R22 and R23, wherein q is 0, 1, 2, 3, or 4;
• or a moiety represented by a formula:
• wherein each of R20a and R20b is independently selected from hydrogen and C1-C4 alkyl;
• and
• wherein each of R21a and R21b is independently C3-C6 alkylene;
• or a moiety represented by a formula:
• wherein each of R4a, R4b, R4c, and R4d is independently selected from hydrogen, hydroxy, amino, C1-C4 alkyl, and —CO2H;
• and
• wherein each of R11 and R12 is independently C3-C6 alkylene;
• wherein each of R22 and R23 is independently selected from:
• hydrogen when adjacent a double bond and hydrogen or —SR24 when adjacent a single bond, provided that both R22 and R23 are not simultaneously hydrogen adjacent a single bond;
• wherein each R24, when present, is independently selected from hydrogen, C1-C24 alkyl, —(CH2CH2O)n(C1-C4 alkyl), a solubilizing functionality, a labeling functionality, a tether to solid-phase support, or a tether to a second peptide;
• and
• wherein each occurrence of n is an integer from 1 to 12,
• or a pharmaceutically acceptable salt thereof.

A compound having a structure represented by a formula:

• wherein each occurrence of is an optional covalent bond, thereby signifying a single bond or a double bond;
• wherein each of j, u, x, and z is independently an integer having a value of 0 to 100;
• provided that j and x are not simultaneously 0;
• and
• provided that u and z are not simultaneously 0;
• wherein each instance of Xaa in [Xaa]x is, independently, a natural or unnatural amino acid residue when x is 1 to 100;
• and wherein [Xaa]x is hydrogen when x is 0;
• wherein each instance of Xaa in [Xaa]y is, independently, a natural or unnatural amino acid;
• wherein each instance of Xaa in [Xaa]z is, independently, a natural or unnatural amino acid when z is 1 to 100;
• and wherein [Xaa]z is hydroxy when z is 0;
• wherein each of R1a and R1b is independently hydrogen or an amino protecting group;
• wherein each of R2a and R2b is independently hydrogen or C1-C4 alkyl;
• wherein each of R3a and R3b is independently a C1-C4 alkylene;
• wherein Z is C5-C12 alkylene, —(C2-C6 alkylene)-O—(C2-C6 alkylene)-, —(CH2)3—(OCH2CH2)q—O—(CH2)3—, wherein q is 0, 1, 2, 3, or 4;
• or a moiety represented by a formula:
• wherein each of R20a and R20b is independently selected from hydrogen and C1-C4 alkyl;
• and
• wherein each of R21a and R21b is independently C3-C6 alkylene;
• or a moiety represented by a formula:
• wherein q is 0, 1, 2, 3, or 4;
• wherein each of R4a, R4b, R4c, and R4d is independently hydrogen, hydroxy, amino, C1-C4 alkyl, and —CO2H;
• and
• wherein each of R11 and R12 is independently C3-C6 alkylene;
• wherein each of R22 and R23 is independently selected from:
• hydrogen when adjacent a double bond and hydrogen or —SR24 when adjacent a single bond, provided that both R22 and R23 are not simultaneously hydrogen adjacent a single bond;
• wherein each R24, when present, is independently selected from hydrogen, C1-C24 alkyl, —(CH2CH2O)n(C1-C4 alkyl), a solubilizing functionality, a labeling functionality, a tether to solid-phase support, or a tether to a second peptide;
• and
• wherein each occurrence of n is an integer from 1 to 12,
• or a pharmaceutically acceptable salt thereof.

A method of preparing a stapled peptide, the method comprising the steps of:

• a) providing a peptide having the structure represented by the formula:
• wherein each of x and z is independently an integer having a value of 0 to 100;
• wherein y is an integer having a value of 1 to 20;
• wherein each instance of Xaa is, independently, a natural or unnatural amino acid;
• wherein each of R1a and R1b is independently hydrogen or an amino protecting group;
• wherein each of R2a and R2b is independently hydrogen or C1-C4 alkyl;
• wherein each of R3a and R3b is independently a C1-C4 alkyl;
• b) providing a linker compound having the structure represented by the formula:
• wherein Z1 is C1-C8 alkyl, C3-C8 ether, C6-C8 polyether,
• or a moiety represented by a formula:
• wherein each of R20a and R20b is independently selected from hydrogen and C1-C4 alkyl;
• and
• wherein each of R21a and R21b is independently C3-C6 alkylene;
• or a moiety represented by a formula:
• wherein each of R4a, R4b, R4c, and R4d is independently hydrogen, hydroxy, amino, C1-C4 alkyl, and —CO2H;
• and
• wherein each of R13 and R14 is independently C1-C4 alkyl;
• c) reacting the peptide and the linker compound;
• thereby forming the stapled peptide.

A method of preparing a stapled peptide, the method comprising the steps of:

• a) providing a first peptide and a second peptide having, respectively, the structure represented by the formulas:
• wherein each of j, u, x, and z is independently an integer having a value of 0 to 100;
• provided that j and x are not simultaneously 0;
• and
• provided that u and z are not simultaneously 0;
• wherein each instance of Xa is, independently, a natural or unnatural amino acid;
• wherein each of R1a and R1b is independently hydrogen or an amino protecting group;
• wherein each of R2a and R2b is independently hydrogen or C1-C4 alkyl;
• wherein each of R3a and R3b is independently a C1-C4 alkyl;
• b) providing a linker compound having the structure represented by the formula:
• wherein Z1 is C1-C8 alkyl, C3-C8 ether, C6-C8 polyether,
• or a moiety represented by a formula:
• wherein each of R20a and R20b is independently selected from hydrogen and C1-C4 alkyl;
• and
• wherein each of R21a and R21b is independently C3-C6 alkylene;
• or a moiety represented by a formula:
• wherein each of R4a, R4b, R4c, and R4d is independently hydrogen, hydroxy, amino, C1-C4 alkyl, and —CO2H;
• and
• wherein each of R13 and R14 is independently C1-C4 alkyl;
• c) reacting the peptide and the linker compound;
• thereby forming the stapled peptide.