Two-residue-extended peptide and method for producing same

Patent metadata

Field	Value
Publication number	US-2025313594-A1
Application number	US-202318866378-A
Country	US
Kind code	A1
Filing date	Apr 10, 2023
Priority date	May 16, 2022
Publication date	Oct 9, 2025
Grant date	—

Abstract

Various two-residue-extended peptides can be produced by reacting a first amino acid or peptide represented by the formula:a halogenating agent, a tertiary amine represented by the formula:an amino acid N-carboxy anhydride (NCA) represented by the formula:and a second amino acid or peptide represented by the formula:

First claim

1 . A method for producing a two-residue-extended peptide represented by formula (6):

wherein R 1 represents a hydrogen atom or an alkyl group; R 4 , R 8 , and R 10 are the same or different and each represents a hydrogen atom or a methyl group; R 2 , R 5 , R 6 , R 9 , and R 11 are the same or different and each represents a side chain of an amino acid residue; R 2 , R 5 , R 6 , R 9 , and R 11 each optionally form a ring together with the adjacent nitrogen atom through the carbon atom to which each is attached; R 3 , R 7 , and R 12 are the same or different and each represents a hydrogen atom or an optionally substituted monovalent organic group; P 1 represents a hydrogen atom, a protecting group for amino groups, a tag, or a solid-phase; P 2 represents a hydrogen atom, a protecting group for carboxy groups, a tag, or a solid-phase; n1 represents an integer of 0 or more, and when n1 is an integer of 2 or more, n1 R 1 s, R 2 s, and R 3 s are optionally the same or different; and n2 represents an integer of 0 or more, and when n2 is an integer of 2 or more, n2 R 10 s, R 11 s, and R 12 s are optionally the same or different;
the method comprising a reaction step of reacting
a first amino acid or peptide represented by formula (1):

wherein R 1 , R 2 , R 3 , R 4 , R 5 , P 1 , and n1 are as defined above,
a halogenating agent, a tertiary amine represented by formula (3):

wherein R 13 , R 14 , and R 15 are the same or different and each represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, or two or more of R 13 , R 14 , and R 15 are optionally linked to form a ring optionally having one or more heteroatoms or substituents,
an amino acid N-carboxy anhydride (NCA) represented by formula (4):

wherein R 6 and R 7 are as defined above, and
a second amino acid or peptide represented by formula (5):

wherein R 8 , R 9 , R 10 , R 11 , R 12 , P 2 , and n2 are as defined above.

2 . The production method according to claim 1 , wherein the reaction step comprises:
(A) a reaction step of reacting the first amino acid or peptide represented by formula (1), the halogenating agent, the tertiary amine represented by formula (3), and the amino acid N-carboxy anhydride (NCA) represented by formula (4); and (B) a reaction step of reacting a reaction mixture obtained in step (A) and the second amino acid or peptide represented by formula (5).

3 . The production method according to claim 2 , wherein step (A) comprises:
(A1) a reaction step of reacting the first amino acid or peptide represented by formula (1), the halogenating agent, and the tertiary amine represented by formula (3); and (A2) a reaction step of reacting a reaction mixture obtained in step (A1) and the amino acid N-carboxy anhydride (NCA) represented by formula (4).

4 . The production method according to claim 1 , wherein the halogenating agent is thionyl represented by formula (2):

wherein Xs are the same or different and each represents a halogen atom.

5 . The production method according to claim 1 , wherein the tertiary amine comprises an aliphatic amine and/or a heterocyclic aromatic amine.

6 . The production method according to claim 1 , wherein the reaction temperature of the reaction step is −80 to 100° C.

7 . The production method according to claim 1 , wherein the reaction time of the reaction step is less than 60 minutes.

8 . The production method according to claim 1 , wherein the reaction step is performed by a flow method.

9 . The production method according to claim 8 , wherein the reaction step is performed by a micro-flow method.

10 . An acylated NCA represented by formula (8):

wherein R 1 represents a hydrogen atom or an optionally substituted monovalent organic group; R 4 represents a hydrogen atom or a methyl group; R 2 , R 5 , and R 6 are the same or different and each represents a side chain of an amino acid residue; R 2 , R 5 , and R 6 each optionally form a ring together with the adjacent nitrogen atom through the carbon atom to which each is attached; R 3 and R 7 each represent a hydrogen atom or an optionally substituted monovalent organic group; P 1 represents a hydrogen atom, a protecting group for amino groups, a tag, or a solid-phase; and n1 represents an integer of 0 or more, and when n1 is an integer of 2 or more, n1 R 1 s to R 3 s are optionally the same or different.

11 . A peptide represented by formula (9):

wherein R 16 , R 17 , and R 18 are the same or different and each represents a side chain of an amino acid residue; R 16 , R 17 , and R 18 each optionally form a ring together with the adjacent nitrogen atom through the carbon atom to which each is attached; P 3 represents a hydrogen atom or a protecting group for amino groups; and P 4 represents a hydrogen atom or a protecting group for carboxy groups.

12 . A peptide consisting of an amino acid sequence represented by SEQ ID NO: 1.

Assignees

• National Univ Corporation Tokai National Higher Education And Research System

Inventors

• Fuse Shinichiro
• Sugisawa Naoto

Classifications

• C07K1/02

  in solution {(C07K1/003, C07K1/006 take precedence)} · CPC title

• C07K5/0815

  with the first amino acid being basic · CPC title

• C07K5/0821

  with the first amino acid being heterocyclic, e.g. His, Pro, Trp · CPC title

• C07K5/0812

  and aromatic or cycloaliphatic · CPC title

• C07K5/081

  the side chain containing O or S as heteroatoms, e.g. Cys, Ser · CPC title