Peptide library and use thereof

Patent metadata

Field	Value
Publication number	US-11319345-B2
Application number	US-202016777719-A
Country	US
Kind code	B2
Filing date	Jan 30, 2020
Priority date	Aug 8, 2012
Publication date	May 3, 2022
Grant date	May 3, 2022

Abstract

Disclosed are compositions and method related to variants of SPINK2 that bind to targets other than an endogenous target of SPINK2. In one embodiment, a peptide is provided that comprises the amino acid sequence SEQ ID NO: 1. In further embodiments, an amino acid sequences encoded by nucleotide positions 4 to 42 and/or nucleotide positions 94 to 189 in the nucleotide sequence of SEQ ID NO: 14 flank the amino terminus and the carboxyl terminus, respectively, of the amino acid sequence. In another embodiment, a peptide is provided that comprises an amino acid sequence derived from the amino acid sequence of SEQ ID NO: 1 in which a conservative substitution, deletion, addition and/or insertion of 1 to 5 (inclusive) amino acids has occurred at amino acids other than the 1st Xaa to the 12th Xaa counting from the amino terminus.

First claim

The invention claimed is:

1. A peptide library comprising a peptide comprising from amino terminus to carboxy terminus:
(a) a first domain with an amino acid sequence as set forth in positions 2-14 of SEQ ID NO:15;
(b) a second domain with an amino acid sequence as set forth in SEQ ID NO:1; and
(c) a third domain with an amino acid sequence as set forth in positions 32-62 of SEQ ID NO:15;
wherein the peptide library has a diversity of at least 1×10 5 .

2. The peptide library according to claim 1 , wherein the library has a diversity of at least 1×10 8 .

3. The peptide library according to claim 1 , wherein each of the 1st Xaa to the 5th Xaa, the 7th Xaa, the 9th Xaa, and the 10th Xaa counting from the amino terminus of SEQ ID NO: 1 is any amino acid other than cysteine and proline.

4. The peptide library according to claim 1 , wherein each of the 6th Xaa and the 8th Xaa counting from the amino terminus of SEQ ID NO: 1 is any amino acid other than cysteine.

5. The peptide library according to claim 1 , wherein the 11th Xaa counting from the amino terminus of SEQ ID NO: 1 is an amino acid selected from the group consisting of tyrosine, serine, phenylalanine, leucine, and threonine.

6. The peptide library according to claim 1 , wherein the 12th Xaa counting from the amino terminus of SEQ ID NO: 1 is an amino acid selected from the group consisting of asparagine, aspartic acid, leucine, lysine, glutamine, alanine, and glutamic acid.

7. The peptide library according to claim 1 , wherein the library comprises a peptide comprising the amino acid sequence represented by any one of SEQ ID NOs: 2 to 9.

8. The peptide library according to claim 1 , wherein the library is a phage display library, a ribosome display library, or a nucleic acid display library.

9. The peptide library according to claim 1 , wherein the peptide binds to a predetermined target molecule that is not an endogenous target of SPINK2.

10. The peptide library according to claim 9 , wherein the predetermined target molecule is derived from a human.

11. The peptide library according to claim 9 , wherein the predetermined target molecule is trypsin and/or acrosin.

12. The peptide library according to claim 1 , wherein:
the 1st Xaa counting from the amino terminus of SEQ ID NO: 1 is an amino acid selected from the group consisting of arginine, methionine, leucine, tryptophan, and serine;
the 2nd Xaa counting from the amino terminus of SEQ ID NO: 1 is an amino acid selected from the group consisting of threonine, arginine, tryptophan, and phenylalanine;
the 3rd Xaa counting from the amino terminus of SEQ ID NO: 1 is an amino acid selected from the group consisting of arginine, histidine, tryptophan, serine, and phenylalanine;
the 4th Xaa counting from the amino terminus of SEQ ID NO: 1 is an amino acid selected from the group consisting of tryptophan, arginine, and leucine;
the 5th Xaa counting from the amino terminus of SEQ ID NO: 1 is an amino acid selected from the group consisting of glycine, arginine, leucine, histidine, tryptophan, methionine, and tyrosine;
the 6th Xaa counting from the amino terminus of SEQ ID NO: 1 is an amino acid selected from the group consisting of asparagine, histidine, proline, lysine, tryptophan, arginine, and aspartic acid;
the 7th Xaa counting from the amino terminus of SEQ ID NO: 1 is an amino acid selected from the group consisting of arginine, phenylalanine, tryptophan, leucine, alanine, and glycine;
the 8th Xaa counting from the amino terminus of SEQ ID NO: 1 is an amino acid selected from the group consisting of threonine, proline, asparagine, and serine;
the 9th Xaa counting from the amino terminus of SEQ ID NO: 1 is an amino acid selected from the group consisting of tryptophan, methionine, tyrosine, and phenylalanine;
the 10th Xaa counting from the amino terminus of SEQ ID NO: 1 is an amino acid selected from the group consisting of glutamine, valine, lysine, methionine, alanine, leucine, and asparagine;
the 11th Xaa counting from the amino terminus of SEQ ID NO: 1 is an amino acid selected from the group consisting of tyrosine, phenylalanine, and leucine; and/or
the 12th Xaa counting from the amino terminus of SEQ ID NO: 1 is lysine.

13. The peptide library according to claim 1 , wherein the first domain and the second domain are joined directly in the peptide.

14. The peptide library according to claim 1 , wherein the first domain and the second domain are separated by one or two arbitrary amino acids.

15. The peptide library according to claim 1 , wherein the second domain and the third domain are joined directly in the peptide.

16. The peptide library according to claim 1 , wherein the second domain and the third domain are separated by one or two arbitrary amino acids.

Assignees

• Daiichi Sankyo Co Ltd

Inventors

• Nishimiya Daisuke
• Hashimoto Ryuji

Classifications

• C12Q1/37

  involving peptidase or proteinase · CPC title

• C12N2510/00

  Genetically modified cells · CPC title

• C12N15/1037

  Screening libraries presented on the surface of microorganisms, e.g. phage display, E. coli display · CPC title

• C40B50/00

  Methods of creating libraries, e.g. combinatorial synthesis · CPC title

• C07K1/047

  Simultaneous synthesis of different peptide species; Peptide libraries · CPC title