Dual-enzyme responsive peptides

Patent metadata

Field	Value
Publication number	US-11879019-B2
Application number	US-201716469452-A
Country	US
Kind code	B2
Filing date	Dec 19, 2017
Priority date	Dec 19, 2016
Publication date	Jan 23, 2024
Grant date	Jan 23, 2024

Abstract

An enzyme-responsive peptide and a method off using such enzyme-responsive peptide are disclosed. An enzyme-responsive peptide, the peptide comprising an amino acid having an α-amino group, an α-carboxylic acid group and a ε-amine group, wherein the ε-amine group is covalently bonded with a first group and the α-carboxylic acid is covalently bonded with a second group.

First claim

We claim:

1. An enzyme-responsive peptide, wherein the enzyme-responsive peptide has a general structure of:

wherein X is selected from O, NH or S; R 1 is an enzyme substrate, wherein R 1 is a protease substrate selected from the group consisting of a chymotrypsin substrate, a papain substrate, a caspase 8 substrate and a caspase 3 substrate; R 2 comprises a reporter molecule or an active ingredient; and R 3 is selected from a protecting group, a H, an amino acid or a peptide, wherein the enzyme-responsive peptide requires digestion by two separate enzymes to cleave a bond between the α-carboxylic acid and X—R 2 to release R 2 , wherein the enzyme-responsive peptide requires digestion by a first enzyme selected from the group consisting of chymotrypsin, a papain, caspase 8 and caspase 3 and by a second enzyme, wherein the second enzyme is trypsin.

2. The enzyme-responsive peptide of claim 1 , wherein X is NH.

3. The enzyme-responsive peptide of claim 1 , wherein R 2 is a reporter molecule.

4. The enzyme-responsive peptide of claim 1 , wherein R 2 is an active ingredient.

5. The enzyme-responsive peptide of claim 1 , wherein R 3 is a protecting group.

6. The enzyme-responsive peptide of claim 1 , wherein the enzyme-responsive peptide requires digestion by the first enzyme and subsequently by the second enzyme.

7. The enzyme-responsive peptide of claim 1 , wherein R 3 comprises cysteine conjugated to a siloxane polymer.

8. The enzyme-responsive peptide of claim 1 , wherein R 3 comprises CAA.

9. The enzyme-responsive peptide of claim 1 , wherein R 1 comprises F, FG, AAF, or DEVD (SEQ ID NO: 7).

10. The enzyme-responsive peptide of claim 8 , wherein the enzyme-responsive peptide comprises the compound of formula

wherein R 4 is H, acetamide, a protecting group, an amino acid, or a peptide and wherein R 5 is H, a thiol protecting group, or a siloxane polymer.

11. The enzyme-responsive peptide of claim 1 , wherein R 2 comprises the reporter molecule or the active ingredient conjugated through a carbamate linker or a thionocarbamate linker.

12. The enzyme-responsive peptide of claim 11 , wherein the enzyme-responsive peptide comprises the compound of formula

wherein R 4 is H, acetamide, a protecting group, an amino acid, or a peptide, wherein R 5 is H, a thiol protecting group, or a siloxane polymer, wherein Z is selected from O and S, and wherein R 6 is the reporter molecule or the active ingredient conjugated through the carbamate linker or the thionocarbamate linker.

13. A polymeric formulation for controlled releasing an active ingredient, the formulation comprising the enzyme-responsive peptide of claim 1 .

14. A method for controlled releasing an active ingredient, wherein the method comprising the steps of
(1) contacting a compound having a general structure of

wherein X is selected from O, NH or S; R 1 is an enzyme substrate, wherein R 1 is a protease substrate selected from the group consisting of a chymotrypsin substrate, a papain substrate, a caspase 8 substrate and a caspase 3 substrate; R 2 comprises an active ingredient; and R 3 is selected from a protecting group, a H, an amino acid or a peptide; with a first enzyme, wherein R 1 is cleaved from the compound upon digestion by the first enzyme to form a second compound with a free ε-amine group; and
(2) contacting the second compound with a second enzyme, wherein the active ingredient is released from the second compound upon digestion of the second enzyme—wherein the enzyme-responsive peptide requires digestion by a first enzyme selected from the group consisting of chymotrypsin, a papain, caspase 8 and caspase 3 and by a second enzyme, wherein the second enzyme is trypsin.

15. The method of claim 14 , wherein R 3 is a protecting group.

16. The method of claim 14 , wherein the first enzyme is a chymotrypsin.

17. The method of claim 14 , wherein the first enzyme is papain.

18. The method of claim 14 , wherein the first enzyme is a caspase 8.

19. The method of claim 14 , wherein the first enzyme is caspase 3.

Assignees

• Univ California

Inventors

• Maynard Heather D
• Boehnke Natalie

Classifications

• C07K5/0215 Primary

  containing natural amino acids, forming a peptide bond via their side chain functional group, e.g. epsilon-Lys, gamma-Glu · CPC title

• A61K47/65

  Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers · CPC title

• C07K7/02

  Linear peptides containing at least one abnormal peptide link · CPC title

• C12P21/06

  produced by the hydrolysis of a peptide bond, e.g. hydrolysate products (preparing foodstuffs by protein hydrolysis A23J3/00) · CPC title

• C12Q1/37

  involving peptidase or proteinase · CPC title