Process for synthesizing cyclic peptide compound

Patent metadata

Field	Value
Publication number	US-9090668-B2
Application number	US-59322108-A
Country	US
Kind code	B2
Filing date	Mar 26, 2008
Priority date	Mar 26, 2007
Publication date	Jul 28, 2015
Grant date	Jul 28, 2015

Abstract

The objective is to provide a novel process for synthesizing a cyclic peptide compound. It is also an objective to provide a novel cyclic peptide compound. The novel process for synthesizing a cyclic peptide compound comprises the steps of: (1) translationally synthesizing a non-cyclic peptide compound having in a molecule a functional group 1 and a functional group 2, which are a pair of functional groups capable of reacting to form a bond, and (2) cyclizing the non-cyclic peptide compound by the reaction of the functional groups 1 and 2 to form a bond. The novel cyclic peptide compound can be synthesized by the process.

First claim

The invention claimed is:

1. A process for synthesizing a cyclic peptide compound, comprising the steps of:
(1) translationally synthesizing a non-cyclic peptide compound comprising non-natural amino acid(s), and containing a functional group 1 and a functional group 2, which are a pair of functional groups capable of reacting to form a bond; and
(2) cyclizing the non-cyclic peptide compound by the reaction of the functional group 1 with the functional group 2 to form a bond between said groups,
wherein a ribozyme capable of catalyzing an aminoacylation reaction of a tRNA with a moderately activated amino acid compound having an aromatic ring is used to translationally synthesize the non-cyclic peptide compound, and
wherein the pair of functional groups 1 and 2 is one of the following pairs (A) to (C):

wherein X 1 is Cl or Br, and Ar is an aromatic ring optionally having a substituent,
wherein the step (1) comprises the substeps of:
(a) providing the ribozyme capable of catalyzing an aminoacylation reaction of a tRNA with a moderately activated amino acid compound having an aromatic ring;
(b) providing an initiator tRNA and an amino acid compound having the functional group 1 for use as a substrate for the acylation reaction by the ribozyme;
(c) performing an aminoacylation of the initiator tRNA with the amino acid compound having the functional group 1 using the ribozyme;
(d) providing a cell-free translation system which contains at least an amino acid compound having the functional group 2 and a tRNA to be aminoacylated with the amino acid compound having the functional group 2, and is free of methionine;
(e) providing a template DNA for forming an mRNA having at desired positions a codon corresponding to an anticodon of the initiator tRNA, and a codon corresponding to an anticodon of the tRNA to be aminoacylated with the amino acid compound having the functional group 2; and
(f) adding the aminoacylated initiator tRNA obtained in the substep (c) and the template DNA to the cell-free translation system to synthesize the non-cyclic peptide compound.

2. A process for synthesizing a cyclic peptide compound, comprising the steps of:
(1) translationally synthesizing a non-cyclic peptide compound comprising non-natural amino acid(s), and containing a functional group 1 and a functional group 2, which are a pair of functional groups capable of reacting to form a bond; and
(2) cyclizing the non-cyclic peptide compound by the reaction of the functional group 1 with the functional group 2 to form a bond between said groups,
wherein a ribozyme capable of catalyzing an aminoacylation reaction of a tRNA with a moderately activated amino acid compound having an aromatic ring is used to translationally synthesize the non-cyclic peptide compound, and
wherein the pair of functional groups 1 and 2 is one of the following pairs (A) to (C):

wherein X 1 is Cl or Br, and Ar is an aromatic ring optionally having a substituent,
wherein the step (1) comprises the substeps of:
(a) providing the ribozyme capable of catalyzing an aminoacylation reaction of a tRNA with a moderately activated amino acid compound having an aromatic ring;
(b) providing an initiator tRNA and an amino acid compound having the functional group 1 for use as a substrate for the acylation reaction by the ribozyme;
(c) performing an aminoacylation of the initiator tRNA with the amino acid compound having the functional group 1 using the ribozyme;
(d) providing a tRNA having an orthogonal relationship to a natural aminoacyl-tRNA synthetase present in a cell-free translation system, and an amino acid compound having the functional group 2, for use as a substrate for the acylation reaction by the ribozyme;
(e) performing an aminoacylation of the tRNA with the amino acid compound having the functional group 2 using the ribozyme;
(f) providing a cell-free translation system free of methionine;
(g) providing a template DNA for forming an mRNA having at desired positions a codon corresponding to an anticodon of the initiator tRNA, and a codon corresponding to an anticodon of the tRNA; and
(h) adding the aminoacylated initiator tRNA obtained in the substep (c), the tRNA aminoacylated in the substep (e) with the amino acid compound having the functional group 2, and the template DNA to the cell-free translation system to synthesize the non-cyclic peptide compound.

3. A process for synthesizing a cyclic peptide compound, comprising the steps of:
(1) translationally synthesizing a non-cyclic peptide compound comprising non-natural amino acid(s), and containing a functional group 1 and a functional group 2, which are a pair of functional groups capable of reacting to form a bond; and
(2) cyclizing the non-cyclic peptide compound by the reaction of the functional group 1 with the functional group 2 to form a bond between said groups,
wherein a ribozyme capable of catalyzing an aminoacylation reaction of a tRNA with a moderately activated amino acid compound having an aromatic ring is used to translationally synthesize the non-cyclic peptide compound, and
wherein the pair of functional groups 1 and 2 is one of the following pairs (A) to (C):

wherein X 1 is Cl or Br, and Ar is an aromatic ring optionally having a substituent,
wherein the step (1) comprises the substeps of:
(a) providing the ribozyme capable of catalyzing an aminoacylation reaction of a tRNA with a moderately activated amino acid compound having an aromatic ring;
(b) providing a tRNA having an orthogonal relationship to a natural aminoacyl-tRNA synthetase present in a cell-free translation system, and an amino acid compound having the functional group 1, for use as a substrate for the acylation reaction by the ribozyme;
(c) performing an aminoacylation of the tRNA with the amino acid compound having the functional group 1 using the ribozyme;
(d) providing a cell-free translation system containing at least an amino acid compound having the functional group 2, a tRNA to be aminoacylated with the amino acid compound having the functional group 2, an initiator tRNA, methionine, and a methionyl-tRNA synthetase;
(e) providing a template DNA for forming an mRNA having at desired positions a codon corresponding to an anticodon of the tRNA to be aminoacylated with the amino acid compound having the functional group 1, and a codon corresponding to an anticodon of the tRNA to be aminoacylated with the amino acid compound having the functional group 2; and
(f) adding the aminoacylated tRNA obtained in the substep (c) and the template DNA to the cell-free translation system to synthesize the non-cyclic peptide compound.

4. A process for synthesizing a cyclic peptide compound, comprising the steps of:
(1) translationally synthesizing a non-cyclic peptide compound comprising non-natural amino acid(s), and containing a functional group 1 and a functional group 2, which are a pair of functional groups capable of reacting to form a bond; and
(2) cyclizing the non-cyclic peptide compound by the reaction of the functional group 1 with the functional group 2 to form a bond between said groups,
wherein a ribozyme capable of catalyzing an aminoacylation reaction of a tRNA with a moderately activated amino acid compound having an aromatic ring is used to translationally synthesize the non-cyclic peptide c

Assignees

• Suga Hiroaki
• Murakami Hiroshi
• Goto Yuki
• Yamagishi Yusuke
• Ashigai Hiroshi
• Sako Yusuke
• Univ Tokyo

Inventors

• Suga Hiroaki
• Murakami Hiroshi
• Goto Yuki
• Yamagishi Yusuke
• Ashigai Hiroshi
• Sako Yusuke

Classifications

• C12N15/67

  General methods for enhancing the expression · CPC title

• C07K7/56 Primary

  the cyclisation not occurring through 2,4-diamino-butanoic acid · CPC title

• C12P21/02

  having a known sequence of two or more amino acids, e.g. glutathione · CPC title

• C07K7/06

  having 5 to 11 amino acids · CPC title

• C07K7/08

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