Protease-resistant peptide ligands

Patent metadata

Field	Value
Publication number	US-10266566-B2
Application number	US-201414904715-A
Country	US
Kind code	B2
Filing date	Jul 15, 2014
Priority date	Jul 15, 2013
Publication date	Apr 23, 2019
Grant date	Apr 23, 2019

Abstract

This invention relates generally to the discovery of novel protease-resistant peptide ligands and uses thereof. Specifically, the present invention provides a protease-resistant peptide with three to twenty amino acids capable of binding a biological and comprising one or more basic amino acid(s) and I or aromatic amino acids, wherein one or more of the amino acids is substituted with a non-naturally occurring amino acid analog.

First claim

What is claimed is:

1. A peptide capable of binding a biological, the peptide comprising:
one or more hexapeptides selected from the group consisting of HWRGWV (SEQ ID NO:1), HYFKFD (SEQ ID NO:2) and HFRRHL (SEQ ID NO:3);
wherein the HWRGWV (SEQ ID NO:1) hexapeptide is selected from the group consisting of HW Met CitGW Met V (SEQ ID NO:4), HWCitGWV (SEQ ID NO:10), HW met CitGW met V (SEQ ID NO:8), and HW met RGW met V (SEQ ID NO:11),
wherein the HYFKFD (SEQ ID NO:2) hexapeptide is selected from the group consisting of HY for CitGW for V (SEQ ID NO:12), HY met F met K met F met D (SEQ ID NO:13), and HY met F met K met 2F met D (SEQ ID NO:7),
wherein the HFRRHL (SEQ ID NO:3) hexapeptide is selected from the group consisting of HF met CitCitHL (SEQ ID NO:5) and HF carb CitCitHL (SEQ ID NO:14),
wherein the one or more hexapeptides comprise at least one non-natural amino acid selected from the group consisting of N in -methyl-tryptophan, N in -formyl-tryptophan, 4-methylphenylalanine, 4-carbamoyl-phenylalanine, O-methyl-tyrosine, e-dimethyl-lysine, and citrulline; and
wherein the peptide is resistant to digestion by a protease.

2. The protease-resistant peptide of claim 1 , wherein the peptide is resistant to digestion by endopeptidases.

3. The protease-resistant peptide of claim 1 , wherein the peptide is resistant to digestion by exopeptidases.

4. The protease-resistant peptide of claim 2 , wherein the endopeptidase is alpha-chymotrypsin.

5. The protease-resistant peptide of claim 2 , wherein the endopeptidase is trypsin.

6. A chromatographic material comprising a solid support, wherein the solid support is coupled to any of the protease-resistant peptides of claim 1 .

7. The protease-resistant peptide of claim 1 , wherein the biological is an antibody that comprises an IgG domain.

8. The chromatographic material of claim 6 , wherein the solid support comprises resin beads.

9. The chromatographic material of claim 8 , wherein the resin beads comprise amino activated polymethacrylate.

10. A method for isolating a biological, the method comprising:
contacting a chromatographic material with a biological-containing preparation under binding conditions in which the biological is capable of binding to a protease-resistant peptide, wherein the protease-resistant peptide comprises one or more hexapeptides selected from the group consisting of HWRGWV (SEQ ID NO:1), HYFKFD (SEQ ID NO:2) and HFRRHL (SEQ ID NO:3), wherein the HWRGWV (SEQ ID NO:1) hexapeptide is selected from the group consisting of HW Met CitGW Met V (SEQ ID NO:4), HWCitGWV (SEQ ID NO:10), HW met CitGW met V (SEQ ID NO:8), and HW met RGW met V (SEQ ID NO:11), wherein the HYFKFD (SEQ ID NO:2) hexapeptide is selected from the group consisting of HY for CitGW for V (SEQ ID NO:12), HY met F met K met F met D (SEQ ID NO:13), and HY met F met K met 2F met D (SEQ ID NO:7), wherein the HFRRHL (SEQ ID NO:3) hexapeptide is selected from the group consisting of HF met CitCitHL (SEQ ID NO:5) and HF carb CitCitHL (SEQ ID NO:14), and wherein the one or more hexapeptides comprise at least one non-natural amino acid selected from the group consisting of Nin-methyl-tryptophan, Nin-formyl-tryptophan, 4-methylphenylalanine, 4-carbamoyl-phenylalanine, O-methyl-tyrosine, e-dimethyl-lysine, and citrulline;
washing the chromatographic material and the bound biological; and
eluting the biological from the chromatographic material so as to isolate the biological.

11. The method of claim 10 , wherein the biological compound is an antibody that comprises an IgG domain.

12. A diagnostic kit for detecting the presence of a biological comprising the chromatographic material of claim 6 , wherein the biological is an antibody that comprises an IgG domain.

Assignees

• Univ North Carolina State

Inventors

• Menegatti Stefano
• Bobay Benjamin G
• Carbonell Ruben G

Classifications

• C07K16/065

  Purification, fragmentation · CPC title

• C07K7/06 Primary

  having 5 to 11 amino acids · CPC title

• C07K1/22

  Affinity chromatography or related techniques based upon selective absorption processes · CPC title

• C07K7/08

  having 12 to 20 amino acids (gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68) · CPC title