IP Library › Granted Patent US 9,701,715

Granted Patent B2

US 9,701,715 · App. 14/438,194 · Granted Jul 11, 2017

Conformationally-constrained kinked endosomal-disrupting peptides

Inventor: Blake R. Peterson (Lawrence, KS)

Assignee: The University of Kansas

C07K7/08 A61K38/10 A61K47/42 A61K47/48246 C12N2310/30 C12N2310/3513 C12N2310/3515

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Quick Facts

Patent No.: US 9,701,715
App. No.: 14/438,194
Granted: Jul 11, 2017
Kind: B2

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Abstract

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, the kinked portion comprising an endosomal-disrupting peptide. The peptide can include a peptide sequence of one of SEQ ID NOs: 1, 5-38, or 40-54 or 61-69. The conformationally-constrained kinked portion can be a majority portion or minority of the peptide. The peptide can include one of Formulae 1-1C, wherein: CC-Peptide includes a peptide that conformationally constrains the ED-KP; Peptide independently includes natural, unnatural, essential or non-essential aromatic, aliphatic, or other amino acids having L or D configuration; ED-KP includes an endosomal-disrupting kinked peptide; Xaa, Xaa1, and Xaa2 are independently one or more natural or non-natural amino acids, essential amino acids, or non-essential amino acids, or derivatives of amino acids having L or D configuration; L1 and L2 are independently linkers; and n1, n2, n3, and n4 are independently 0-50.

Claims (112)

1. A cell-targeting compound comprising:

one or more of the 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 one of SEQ ID NOs: 5-38, 40-54 or 61-69; and

a targeting moiety linked to an end of the peptide, wherein:

the kinked portion has an amino acid sequence that includes one or more amino acids independently selected from proline and glycine;

the conformationally-constrained portion includes one or more 2-aminoisobutyric acid residues; and

the peptide linker includes 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 or peptide thereof.

2. The cell-targeting compound of claim 1 , wherein the targeting moiety is on the C-terminus of the peptide.

3. The cell-targeting compound of claim 1 , wherein the targeting moiety is on the N-terminus of the peptide.

4. The cell-targeting compound of claim 1 , comprising one of Formulae 2-2C, 3-3C, or 4-4C, wherein:

ED-KP is the 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) 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 .

5. The cell-targeting compound of claim 1 , comprising one of Formulae 6-6C, 7-7C, or 8-8C, wherein:

ED-KP is the 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 .

6. The cell-targeting compound of claim 1 , comprising one of Formulae 10-10C, 11-11C, or 12-12C wherein:

KP and KP1 one or more amino acids independently selected from proline and glycine that can cause the endosomal-disrupting kinked 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 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 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 .

7. The cell-targeting compound of claim 6 , wherein the targeting moiety is on the C-terminus or on N-terminus.

8. The cell-targeting compound of claim 6 , wherein Z 1 and Z 2 are independently a targeting moiety or cargo moiety, wherein at least one is a targeting moiety.

9. The cell-targeting compound of claim 6 , wherein Y 1 and Y 2 are each a linker.

10. The cell-targeting compound of claim 6 , X 1 and X 2 are each independently one or more beta-alanine residues.

11. The cell-targeting compound of claim 6 , wherein Y 1 and Y 2 are each a linker and X 1 and X 2 are each independently one or more beta-alanine residues.

12. The cell-targeting compound of claim 11 , wherein Y 1 and Y 2 are each a linker that includes 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 or peptide thereof.

13. The cell-targeting compound of claim 6 , wherein: X 1 -(Aib) n1 -(Xaa 1 ) n2 -(KP) n3 -(Xaa 2 ) n4 includes one of SEQ ID NOs: 9-38, 41-54 or 61-69.

14. The cell-targeting compound of claim 6 , wherein n1 is 4.

15. The cell-targeting compound of claim 1 , further comprising:

a cargo moiety linked to the peptide.

16. The cell-targeting compound of claim of claim 15 , comprising the cargo moiety linked at an internal portion of the cell-targeting compound between the targeting moiety and the peptide.

17. The cell-targeting compound of claim of claim 15 , wherein the cargo moiety is a therapeutic agent, pharmaceutical, nutraceutical, diagnostic agent, assay agent, tracking agent, suicide agent, toxin, or any other agent.

18. A cell-targeting compound comprising one of SEQ ID NOs: 5-38, 40-54 or 61-69 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 endosomal-disrupting kinked 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 is an integer greater than or equal to 2 and less than or equal to 50;

n3 is 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 ) n1 -(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 ;