Targeted conformationally-constrained kinked endosomal disrupting peptides
A peptide can have a sequence of one of SEQ ID NOs: 78-91. A conformationally-constrained kinked peptide includes: a conformationally-constraining portion and a kinked portion linked to the conformationally-constraining portion that conformationally constrains the kinked portion having a peptide sequence of one of SEQ NOs: 78-97. A cell-targeting compound can include a conformationally-constrained kinked peptide having a peptide sequence of one of SEQ ID NOs: 78-97. The peptide sequence can be one of SEQ ID NOs: 78-97, or 78-91, or 92-97. A cell-targeting compound can include a conformationally-constrained kinked peptide linked to a branched linker with one branch arm linked to a specific targeting moiety and one branch arm linked to a general targeting moiety. The specific targeting moiety can be an antibody. The general targeting moiety can be a lipid or cholesterol derivative.
1. A peptide sequence comprising:
one of SEQ ID NOs: 92-97.
2. A peptide sequence, comprising:
one of SEQ ID NOs: 5-38, 40-54, 61-69, or 78-91.
3. The peptide sequence of claim 2 , comprising:
one of SEQ ID NOs: 78-91.
4. A cell-targeting compound comprising:
one or more peptides having a conformationally-constraining portion, and a kinked portion linked through a peptide linker to the conformationally-constraining portion that conformationally constrains the kinked portion, the kinked portion, peptide linker and conformationally-constraining portion includes the peptide sequence of claim 1 ; and
at least one targeting moiety linked to an end of the one or more peptides.
5. The cell-targeting compound of claim 4 , wherein at least one targeting moiety is on the C-terminus of the peptide.
6. The cell-targeting compound of claim 4 , wherein at least one targeting moiety is on the N-terminus of the peptide.
7. The cell-targeting compound of claim 4 , comprising one of Formulae 2-2C, 3-3C, or 4-4C, wherein:
ED-KP is an endosomal-disrupting kinked peptide having one or more amino acids independently selected from proline and glycine;
CC-Peptide includes a peptide having one or more 2-aminoisobutyric acid residues that conformationally-constrains the ED-KP;
Peptide independently includes natural, unnatural, essential or non-essential aromatic, aliphatic, or other amino acids, or having L or D configuration;
Xaa, Xaa 1 , and Xaa 2 are independently one or more natural or non-natural amino acids, essential amino acids, or non-essential amino acids, or amino acids having L or D configuration;
L1 and L2 are independently linkers;
n1 and n3 are independently an integer greater than 0 and less than 50;
n2 and n4 are independently 0-50;
Z 1 and Z 2 are independently a targeting moiety, cargo moiety, or nothing, wherein at least one is a targeting moiety;
Y 1 and Y 2 are independently nothing or a linker, or a linker having a cargo moiety; and
X 1 and X 2 are independently nothing, a coupling group, one or more beta-alanine residues, or a polypeptide,
Formula 2=Z 1 —Y 1 —X 1 —(CC-Peptide) n1 -(Peptide) n2 -(ED-KP) n3 -(Peptide) n4 -X 2 —Y 2 —Z 2 ;
Formula 2A =Z 1 —Y 1 —X 1 —(CC-Peptide) n1 -(L1) n2 -(ED-KP) n3 -(L2) n4 -X 2 —Y 2 —Z 2 ;
Formula 2B =Z 1 —Y 1 —X 1 —(CC-Peptide) n1 -(Xaa) n2 -(ED-KP) n3 -(Xaa) n4 -X 2 —Y 2 —Z 2 ;
Formula 2C =Z 1 —Y 1 —X 1 —(CC-Peptide) n1 -(Xaa 1 ) n2 -(ED-KP) n3 -(Xaa 2 ) n4 -X 2 —Y 2 —Z 2 ;
Formula 3=Z 1 —Y 1 —X 1 —(CC-Peptide) n1 -(Peptide) n2 -(ED-KP) n3 -(Peptide) n4 ;
Formula 3A =Z 1 —Y 1 —X 1 —(CC-Peptide) n1 -(L1) n2 -(ED-KP) n3 -(L2) n4 ;
Formula 3B =Z 1 —Y 1 —X 1 —(CC-Peptide) n1 -(Xaa) n2 -(ED-KP) n3 -(Xaa) n4 ;
Formula 3C =Z 1 —Y 1 —X 1 —(CC-Peptide) n1 -(Xaa 1 ) n2 -(ED- KP) n3 -(Xaa 2 ) n4 ;
Formula 4=(CC-Peptide) n1 -(Peptide) n2 -(ED-KP) n3 -(Peptide) n4 -X 2 —Y 2 —Z 2 ;
Formula 4A =(CC-Peptide) n1 -(L1) n2 -(ED-KP) n3 -(L2) 4 -X 2 —Y 2 —Z 2 ;
Formula 4B =(CC-Peptide) n1 -(Xaa) n2 -(ED-KP) n3 -(Xaa) n4 -X 2 —Y 2 —Z 2 ; and
Formula 4C =(CC-Peptide) n1 -(Xaa 1 ) n2 -(ED-KP) n3 -(Xaa 2 ) n4 -X 2 —Y 2 —Z 2 .
8. The cell-targeting compound of claim 4 , comprising one of Formulae 6-6C, 7-7C, or 8-8C, wherein:
ED-KP is an endosomal-disrupting kinked peptide having one or more amino acids independently selected from proline and glycine;
CCM includes a moiety having one or more 2-aminoisobutyric acid residues that conformationally constrains the ED-KP;
Peptide independently includes natural, unnatural, essential or non-essential aromatic, aliphatic, or other amino acids, or having L or D configuration;
Xaa, Xaa 1 , and Xaa 2 are independently one or more natural or non-natural amino acids, essential amino acids, or non-essential amino acids, or amino acids having L or D configuration;
L 1 and L2 are independently linkers;
n1 and n3 are independently an integer greater than 0 and less than 50;
n2 and n4 are independently 0-50;
Z 1 and Z 2 are independently a targeting moiety, cargo moiety, or nothing, wherein at least one is a targeting moiety;
Y 1 and Y 2 are independently nothing or a linker, or a linker having a cargo moiety; and
X 1 and X 2 are independently nothing, a coupling group, one or more beta-alanine residues, or a polypeptide,
Formula 6=Z 1 —Y 1 —X 1 —(CCM) n1 -(Peptide) n2 -(ED-KP) n3 -(Peptide) n4 -X 2 —Y 2 —Z 2 ;
Formula 6A =Z 1 —Y 1 —X 1 —(CCM) n1 -(L1) n2 -(ED-KP) n3 -(L2) n4 -X 2 —Y 2 —Z 2 ;
Formula 6B =Z 1 —Y 1 —X 1 —(CCM) n1 -(Xaa) n2 -(ED-KP) n3 -(Xaa) n4 -X 2 —Y 2 —Z 2 ;
Formula 6C =Z 1 —Y 1 —X 1 —(CCM) n1 -(Xaa 1 ) n2 -(ED-KP) n3 -(Xaa 2 ) n4 -X 2 —Y 2 —Z 2 ;
Formula 7=Z 1 —Y 1 —X 1 —(CCM) n1 -(Peptide) n2 -(ED-KP) n3 -(Peptide) n4 ;
Formula 7A =Z 1 —Y 1 —X 1 —(CCM) n1 -(L1) n2 -(ED-KP) n3 -(L2) n4 ;
Formula 7B =Z 1 —Y 1 —X 1 —(CCM) n1 -(Xaa) n2 -(ED-KP) n3 -(Xaa) n4 ;
Formula 7C =Z 1 —Y 1 —X 1 —CCM) n1 -(Xaa 1 ) n2 -(ED-KP) n3 -(Xaa 2 ) n4 ;
Formula 8=(CCM) n1 -(Peptide) n2 -(ED-KP) n3 -(Peptide) n4 -X 2 —Y 2 —Z 2 ;
Formula 8A =(CCM) n1 -(L1) n2 -(ED-KP) n3 -(L2) n4 -X 2 —Y 2 —Z 2 ;
Formula 8B =(CCM) n1 -(Xaa) n2 -(ED-KP) n3 -(Xaa) n4 -X 2 —Y 2 —Z 2 ; and
Formula 8C =(CCM) n1 -(Xaa 1 ) n2 -(ED-KP) n3 -(Xaa 2 ) n4 -X 2 —Y 2 —Z 2 .
9. The cell-targeting compound of claim 4 , comprising one of Formulae 10-10C, 11-11C, or 12-12C wherein:
KP and KP1 are independently one or more amino acids independently selected from proline and glycine that can cause the peptide to kink;
Aib is a 2-aminoisobutyric acid residue;
Xaa, Xaa 1 , and Xaa 2 are independently one or more natural or non-natural amino acids, essential amino acids, or non-essential amino acids, or amino acids having L or D configuration;
Z 1 and Z 2 are independently a targeting moiety, cargo moiety, or nothing, wherein at least one is a targeting moiety;
Y 1 and Y 2 are independently nothing or a linker, or a linker having a cargo moiety;
X 1 and X 2 are independently nothing, a coupling group, one or more beta-alanine residues, or a polypeptide; and
n1 and n3 are independently an integer greater than 0 and less than or equal to 50;
n2 n4, n5, n6, and n7 are independently 0-50,
Formula 10=Z 1 —Y 1 —X 1 —(Aib) n1 -(Xaa 1 ) n2 -(KP) n3 -(Xaa 2 ) n4 -X 2 —Y 2 —Z 2 ;
Formula 11=Z 1 —Y 1 —X 1 —(Aib) n1 -(Xaa l ) n2 -(KP) n3 -(Xaa 2 ) n4 ;
Formula 12=(Aib) n1 -(Xaa 1 ) n2 -(KP) n3 -(Xaa 2 ) n4 -X 2 —Y 2 —Z 2 ;
Formula 10A =Z 1 —Y 1 —X 1 —(Aib) n1 -(Xaa 1 ) n2 -(KP) n3 -(Xaa 2 ) n4 -(KP1) n5 -X 2 —Y 2 —Z 2 ;
Formula 11A =Z 1 —Y 1 —X 1 —(Aib) n1 -(Xaa 1 ) n2 -(KP) n3 -(Xaa 2 ) n4 -(KP1) n5 ;
Formula 12A =(Aib) n1 -(Xaa 1 ) n2 -(KP) n3 -(Xaa 2 ) n4 -(KP1) n5 -X 2 —Y 2 —Z 2 ;
Formula 10B =Z 1 —Y 1 —X 1 —(Aib) n1 -(Xaa 1 ) n2 -(KP) n3 -(Xaa 2 ) n4 -(KP1) n5 -(Xaa 3 ) n6 -X 2 —Y 2 —Z 2 ;
Formula 11B =Z 1 —Y 1 —X 1 —(Aib) n1 -(Xaa 1 ) n2 -(KP) n3 -(Xaa 2 ) n4 -(KP1) n5 -(Xaa 3 ) n6 ;
Formula 12B =(Aib) n1 -(Xaa 1 ) n2 -(KP) n3 -(Xaa 2 ) n4 -(KP1) n5 -(Xaa 3 ) n6 -X 2 —Y 2 —Z 2 ;
Formula 10C =Z 1 —Y 1 —X 1 —(Xaa 4 ) n7 -(Aib) n1 -(Xaa 1 ) n2 -(KP) n3 -(Xaa 2 ) n4 -(KP1) n5 -X 2 —Y 2 —Z 2 ;
Formula 11C =Z 1 —Y 1 —X 1 —(Xaa 4 ) n7 -(Aib) n1 -(Xaa l ) n2 -(KP) n3 -(Xaa 2 ) n4 -(KP1) n5 ; and
Formula 12C =(Xaa 4 ) 7 -(Aib) n1 -(Xaa 1 ) n2 -(KP) n3 -(Xaa 2 ) n4 -(KP1) n5 -X 2 —Y 2 —Z 2 .
10. The cell-targeting compound of claim 4 , further comprising:
at least one cargo moiety linked to the one or more peptides.
11. The cell-targeting compound of claim 4 , wherein the at least one targeting moiety is linked to the one or more peptides through a branched linker.
12. The cell-targeting moiety of claim 11 , wherein a first arm of the branched linker is linked to a specific targeting moiety and a second arm of the branched linker is linked to a general targeting moiety.
13. The cell targeting moiety of claim 12 , wherein the specific targeting moiety is specific to a protein or portion thereof that is associated with a cell membrane and the general targeting moiety generally associates with the cell membrane.
14. The cell targeting moiety of claim 13 , wherein the specific targeting moiety is an antibody, or fragment thereof, and the general targeting moiety is a lipid or a cholesterol or cholesterol derivative.
15. A cell-targeting compound comprising:
one or more peptides having a conformationally-constraining portion, and a kinked portion linked through a peptide linker to the conformationally-constraining portion that conformationally constrains the kinked portion, the kinked portion, peptide linker and conformationally-constraining portion includes the peptide sequence of claim 3 ; and
at least one targeting moiety linked to an end of the one or more peptides.
16. The cell-targeting compound of claim 15 , wherein at least one targeting moiety is on the C-terminus of the peptide.
17. The cell-targeting compound of claim 15 , wherein at least one targeting moiety is on the N-terminus of the peptide.
18. The cell-targeting compound of claim 15 , comprising one of Formulae 2-2C, 3-3C, or 4-4C, wherein:
ED-KP is an endosomal-disrupting kinked peptide having one or more amino acids independently selected from proline and glycine;
CC-Peptide includes a peptide having one or more 2-aminoisobutyric acid residues that conformationally-constrains the ED-KP;
Peptide independently includes natural, unnatural, essential or non-essential aromatic, aliphatic, or other amino acids, or having L or D configuration;
Xaa, Xaa 1 , and Xaa 2 are independently one or more natural or non-natural amino acids, essential amino acids, or non-essential amino acids, or amino acids having L or D configuration;
L1 and L2 are independently linkers;
n1 and n3 are independently an integer greater than 0 and less than 50;
n2 and n4 are independently 0-50;
Z 1 and Z 2 are independently a targeting moiety, cargo moiety, or nothing, wherein at least one is a targeting moiety;
Y 1 and Y 2 are independently nothing or a linker, or a linker having a cargo moiety; and
Y 1 and X 2 are independently nothing, a coupling group, one or more beta-alanine residues, or a polypeptide,
Formula 2 =Z 1 —Y 1 —X 1 —(CC-Peptide) n1 -(Peptide) n2 -(ED-KP) n3 -(Peptide) n4 -X 2 —Y 2 —Z 2 ;
Formula 2A =Z 1 —Y 1 —X 1 —(CC-Peptide) 1 -(L1) n2 -(ED-KP) n3 -(L2)-X 1 —Y 2 —Z 2 ;
Formula 2B =Z 1 —Y 1 —X 1 —(CC-Peptide) n1 -(Xaa) n2 -(ED-KP) n3 -(Xaa) n4 -X 2 —Y 2 —Z 2 ;
Formula 2C =Z 1 —Y 1 —X 1 —(CC-Peptide) n1 -(Xaa 1 ) n2 -(ED-KP) n3 -(Xaa 2 ) n4 -X 2 —Y 2 —Z 2 ;
Formula 3=Z 1 —Y 1 —X 1 —(CC-Peptide) n1 -(Peptide) n2 -(ED-KP) n3 -(Peptide) n4 ;
Formula 3A =Z 1 —Y 1 —X 1 —(CC-Peptide) n1 -(L1) n2 -(ED-KP) n3 -(L2) n4 ;
Formula 3B =Z 1 —Y 1 —X 1 —(CC-Peptide) n1 -(Xaa) n2 -(ED-KP) n3 -(Xaa) n4 ;
Formula 3C =Z 1 —Y 1 —X 1 —(CC-Peptide) n1 -(Xaa 1 ) n2 -(ED-KP) n3 -(Xaa 2 ) n4 ;
Formula 4=(CC-Peptide) n1 -(Peptide) n2 -(ED-KP) n3 -(Peptide) n4 -X 2 —Y 2 —Z 2 ;
Formula 4A =(CC-Peptide) n1 -(L1) n2 -(ED-KP) n3 -(L1) n4 -X 2 —Y 2 —Z 2 ;
Formula 4B =(CC-Peptide) n1 -(Xaa) n2 -(ED-KP) n3 -(Xaa) n4 -X 2 —Y 2 —Z 2 ; and
Formula 4C =(CC-Peptide) n1 -(Xaa 1 ) n2 -(ED-KP) n3 -(Xaa 2 ) n4 -X 2 —Y 2 —Z 2 .
19. The cell-targeting compound of claim 15 , comprising one of Formulae 6-6C, 7-7C, or 8-8C, wherein:
ED-KP is an endosomal-disrupting kinked peptide having one or more amino acids independently selected from proline and glycine;
CCM includes a moiety having one or more 2-aminoisobutyric acid residues that conformationally constrains the ED-KP;
Peptide independently includes natural, unnatural, essential or non-essential aromatic, aliphatic, or other amino acids, or having L or D configuration;
Xaa, Xaa 1 , and Xaa 2 are independently one or more natural or non-natural amino acids, essential amino acids, or non-essential amino acids, or amino acids having L or D configuration;
L1 and L2 are independently linkers;
n1 and n3 are independently an integer greater than 0 and less than 50;
n2 and n4 are independently 0-50;
Z 1 and Z 2 are independently a targeting moiety, cargo moiety, or nothing, wherein at least one is a targeting moiety;
Y 1 and Y 2 are independently nothing or a linker, or a linker having a cargo moiety; and
X 1 and X 2 are independently nothing, a coupling group, one or more beta-alanine residues, or a polypeptide,
Formula 6 =Z 1 —Y 1 —X 1 —(CCM) n1 -(Peptide) n2 -(ED-KP) n3 -(Peptide) n4 -X 2 —Y 2 —Z 2 ;
Formula 6A =Z 1 —Y 1 —X 1 —(CCM) n1 -(L1) n2 -(ED-KP) n3 -(L2) n4 -X 2 —Y 2 —Z 2 ;
Formula 6B =Z 1 —Y 1 —X 1 —(CCM) n1 -(Xaa) n2 -(ED-KP) n3 -(Xaa) n4 -X 2 —Y 2 —Z 2 ;
Formula 6C =Z 1 —Y 1 —X 1 —(CCM) n1 -(Xaa 1 ) n2 -(ED-KP) n3 -(Xaa 2 ) n4 -X 2 —Y 2 —Z 2 ;
Formula 7=Z 1 —Y 1 —X 1 —(CCM) n1 -(Peptide) n2 -(ED-KP) n3 -(Peptide) n4 ;
Formula 7A =Z 1 —Y 1 —X 1 —(CCM) n1 -(L1) n2 -(ED-KP) n3 -(L2) n4 ;
Formula 7B =Z 1 —Y 1 —X 1 —(CCM) n1 -(Xaa) n2 -(ED-KP) n3 -(Xaa) n4 ;
Formula 7C =Z 1 —Y 1 —X 1 -CCM) n1 -(Xaa 1 ) n2 -(ED-KP) n3 -(Xaa 2 ) n4 ;
Formula 8 =(CCM) n1 -(Peptide) n2 -(ED-KP) n3 -(Peptide) n4 -X 2 —Y 2 —Z 2 ;
Formula 8A =(CCM) n1 -(L1) n2 -(ED-KP) n3 -(L2) n4 -X 2 —Y 2 —Z 2 ;
Formula 8B =(CCM) n1 -(Xaa) n2 -(ED-KP) n3 -(Xaa) n4 -X 2 —Y 2 —Z 2 ; and
Formula 8C =(CCM) n1 -(Xaa 1 ) n2 -(ED-KP) n3 -(Xaa 2 ) n4 -X 2 —Y 2 —Z 2 .
20. The cell-targeting compound of claim 15 , comprising one of Formulae 10-10C, 11-11C, or 12-12C wherein:
KP and KP1 are independently one or more amino acids independently selected from proline and glycine that can cause the peptide to kink;
Aib is a 2-aminoisobutyric acid residue;
Xaa, Xaa 1 , and Xaa 2 are independently one or more natural or non-natural amino acids, essential amino acids, or non-essential amino acids, or amino acids having L or D configuration;
Z 1 and Z 2 are independently a targeting moiety, cargo moiety, or nothing, wherein at least one is a targeting moiety;
Y 1 and Y 2 are independently nothing or a linker, or a linker having a cargo moiety;
X 1 and X 2 are independently nothing, a coupling group, one or more beta-alanine residues, or a polypeptide; and
n1 and n3 are independently an integer greater than 0 and less than or equal to 50;
n2 n4, n5, n6, and n7 are independently 0-50,
Formula 10 =Z 1 —Y 1 —X 1 —(Aib) n1 -(Xaa 1 ) n2 -(KP) n3 -(Xaa 2 ) n4 -X 2 —Y 2 —Z 2 ;
Formula 11 =Z 1 —Y 1 —X 1 —(Aib) n1 -(Xaa 1 ) n2 -(KP) n3 -(Xaa 2 ) n4 ;
Formula 12 =(Aib) n1 -(Xaa 1 ) n2 -(KP) n3 -(Xaa 2 ) n4 -X 2 —Y 2 —Z 2 ;
Formula 10A =Z 1 —Y 1 —X 1 —(Aib) n1 -(Xaa 1 ) n2 -(KP) n3 -(Xaa 2 ) n4 -(KP1) n5 -X 2 —Y 2 —Z 2 ;
Formula 11A =Z 1 —Y 1 —X 1 —(Aib) n1 -(Xaa l ) n2 -(KP) n3 -(Xaa 2 ) n4 -(KP1) n5 ;
Formula 12A =(Aib) n1 -(Xaa 1 ) n2 -(KP) n3 -(Xaa 2 ) n4 -(KP1) n5 -X 2 —Y 2 —Z 2 ;
Formula 10B =Z 1 —Y 1 —X 1 —(Aib) n1 -(Xaa 1 ) n2 -(KP) n3 -(Xaa 2 ) n4 -(KP1) n5 -(Xaa 3 ) n6 -X 2 —Y 2 —Z 2 ;
Formula 11B =Z 1 —Y 1 —X 1 —(Aib) n1 -(Xaa 1 ) n2 -(KP) n3 -(Xaa 2 ) n4 -(KP1) n5 -(Xaa 3 ) n6 ;
Formula 12B =(Aib) n1 -(Xaa 1 ) n2 -(KP) n3 -(Xaa 2 ) n4 -(KP1) n5 -(Xaa 3 ) n6 -X 2 —Y 2 —Z 2 ;
Formula 10C =Z 1 —Y 1 —X 1 —(Xaa 4 ) n7 -(Aib) n1 -(Xaa 1 ) n2 -(KP) n3 -(Xaa 2 ) n4 -(KP1) n5 -X 2 —Y 2 —Z 2 ;
Formula 11C =Z 1 —Y 1 —X 1 —(Xaa 4 ) n7 -(Aib) n1 -(Xaa 1 ) n2 -(KP) n3 -(Xaa 2 ) n4 -(KP1) n5 ; and
Formula 12C =(Xaa 4 ) n7 -(Aib) n1 -(Xaa 1 ) n2 -(KP) n3 -(Xaa 2 ) n4 -(KP1) n5 -X 2 —Y 2 —Z 2 .
21. The cell-targeting compound of claim 15 , further comprising:
at least one cargo moiety linked to the one or more peptides.
22. The cell-targeting compound of claim 15 , wherein the at least one targeting moiety is linked to the one or more peptides through a branched linker.
23. The cell-targeting moiety of claim 22 , wherein a first arm of the branched linker is linked to a specific targeting moiety and a second arm of the branched linker is linked to a general targeting moiety.
24. The cell targeting moiety of claim 23 , wherein the specific targeting moiety is specific to a protein or portion thereof that is associated with a cell membrane and the general targeting moiety generally associates with the cell membrane.
25. The cell targeting moiety of claim 24 , wherein the specific targeting moiety is an antibody, or fragment thereof, and the general targeting moiety is a lipid or cholesterol derivative.