Polypeptide conjugates for intracellular delivery of stapled peptides

1. A polypeptide conjugate comprising a cyclic cell penetrating peptide (cCPP) and a stapled peptide having a structure according to Formula IA:

wherein:

the stapled peptide comprises U, Y 1 , Y 2 , X, Z, J, Z′ or a combination thereof and a staple;

each of X and Z, at each instance, are independently an amino acid;

U, at each instance and when present, is independently an amino acid;

a is a number in the range of from 0 to 500;

c is 3, 6, or 10;

d is a number in the range of from 1 to 500;

Y 1 is an amino acid which has a side chain which forms a first bonding group (b 1 ) to the staple;

Y 2 is an amino acid which has a side chain which forms a second bonding group (b2) to the staple;

cCPP is a cyclic peptide comprising about 4 to about 13 amino acids, wherein the about 4 to about 13 amino acids include at least two arginines and at least two amino acids with hydrophobic side chains;

wherein:

the staple comprises an amide, alkylene substituted with an oxo or N-oxide, or an aryl;

the linker comprises at least one amino acid, alkylene, alkenylene, alkynylene, aryl, cycloalkyl, cycloalkenyl, cycloalkynyl, heterocyclyl, heteroaryl, ether, or a combination thereof, each of which are optionally substituted; and

each of b1 and b2 are independently thioether, disulfide, amide, ester, or ether.

2. The polypeptide conjugate of claim 1 , wherein the cCPP has a sequence comprising any of Formula IIIA-D:

wherein:

each of AA H1 and AA H2 are independently a D or L hydrophobic amino acid;

at each instance and when present, each of AA u and AA z are independently a D or L amino acid; and

wherein the sum of m and n is from 2 to 6.

3. The polypeptide conjugate of claim 1 , wherein c is 3.

4. The polypeptide conjugate of claim 1 , wherein:

(i) U is present, a is 1, and U is either the N-terminus or the C-terminus of the stapled peptide; or

(ii) U is present, a is 2 or more, and the terminal U is either the N-terminus or the C-terminus of the stapled peptide.

5. A cell comprising the polypeptide conjugate of claim 1 .

6. A method for cellular delivery of a stapled peptide, the method comprising contacting a cell with the polypeptide conjugate of claim 1 .

7. A method for making the polypeptide conjugate of claim 1 , the method comprising conjugating a stapled peptide and a cCPP.

8. A method for making a polypeptide conjugate of claim 1 , the method comprising conjugating a peptide to at least one cCPP, and stapling the peptide.

9. A pharmaceutical composition comprising the polypeptide conjugate of claim 1 .

10. The polypeptide conjugate of claim 1 , comprising any one of peptide 4, peptide 11, peptide 13, peptide 15, peptide 17, peptide 19, peptide 21, or peptide 23:

11. The polypeptide conjugate of claim 1 having the following structure:

12. The polypeptide conjugate of claim 1 , wherein the linker has a structure L-1 or L-2:

wherein:

AA s is a side chain or terminus of an amino acid on the peptide or staple;

AA c is a side chain or terminus of an amino acid of the cCPP;

p is an integer from 0 to 10; and

q is an integer from 1 to 50.

13. The polypeptide conjugate of claim 1 , wherein the linker is capable of releasing the stapled peptide from the cCPP after the polypeptide conjugate enters the cytosol of a cell.

14. The polypeptide conjugate of claim 13 , wherein the linker is covalently bound to the stapled peptide through a disulfide bond.