Polypeptide conjugates for intracellular delivery of stapled peptides

Patent metadata

Field	Value
Publication number	US-11510991-B2
Application number	US-201816759600-A
Country	US
Kind code	B2
Filing date	Oct 28, 2018
Priority date	Oct 27, 2017
Publication date	Nov 29, 2022
Grant date	Nov 29, 2022

Abstract

The present disclosure provides novel polypeptide conjugates. The polypeptide conjugates disclosed herein comprise a stapled peptide comprising a peptide and at least one staple which holds the peptide in an α-helical conformation, and a cyclic cell-penetrating peptide (cCPP) conjugated, directly or indirectly, to the stapled peptide. The present disclosure demonstrates that cCPPs can be used to confer consistent cell-permeability to stapled peptides.

First claim

What is claimed is:

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

wherein:
the stapled peptide comprises U, Y 1 , Y 2 , X, Z, J, Z′ or a combination thereof and a staple and contains at least one region having an alpha-helical structure;
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;
J, at each instance and when present, is independently selected from an amino acid;
Z′, 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;
e is a number in the range of from 0 to 500; each of g and h are independently and at each instance 0 or 1, provided that in at least one instance g is 1;
i is a number in the range of from 0 to 100;
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 absent, and Z′ is either the N-terminus or the C-terminus of the stapled peptide;
(ii) U is present, a is 1, and U is either the N-terminus or the C-terminus of the stapled peptide; or
(iii) 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.

Assignees

• Ohio State Innovation Foundation

Inventors

• Pei Dehua

Classifications

• A61K38/00

  Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• C07K14/001 Primary

  by chemical synthesis · CPC title

• C07K7/52

  with only normal peptide links in the ring · CPC title

• C07K14/4703

  Inhibitors; Suppressors · CPC title