Compound library and method for producing compound library

Patent metadata

Field	Value
Publication number	US-12359237-B2
Application number	US-201917279634-A
Country	US
Kind code	B2
Filing date	Sep 27, 2019
Priority date	Sep 28, 2018
Publication date	Jul 15, 2025
Grant date	Jul 15, 2025

Abstract

The present invention provides a method for producing a compound library comprising two or more cyclic compounds represented by the formula (I), comprising a step of allowing a macrocyclase in vitro to act on two or more peptides represented by the formula (II): LP-X-(Xa) m -Y-Z (II) wherein X represents a group represented by the formula (1), Y is a peptide residue consisting of four amino acids and/or analogs thereof and contains a group represented by the formula (2) (wherein R 1 and B 1 are as defined above, and R 3 represents a hydrogen or a hydrocarbon group), and LP is present or absent and, when present, represents a peptide residue consisting of 1 to 100 amino acids and/or analogs thereof, and forming the nitrogen-containing 6-membered ring A while eliminating LP, if present, to form the two or more cyclic compounds represented by the formula (I).

First claim

The invention claimed is:

1. An in vitro method for producing a compound library comprising two or more cyclic compounds represented by the formula (I):

wherein
m number of X a , and X b and X c each independently represent an amino acid residue selected from the group consisting of any amino acids and analogs thereof,
Z is present or absent and, when present, represents a peptide consisting of 1 to 100 amino acids and/or analogs thereof,
m is an integer selected from 2 to 40,
ring A is a nitrogen-containing 6-membered ring optionally having a substituent,
B 1 is an oxygen atom, a sulfur atom, or a NH group, and
R 1 is a hydrogen atom or a hydrocarbon group,

the method comprising allowing a macrocyclase in vitro to act on two or more peptides represented by the formula (II):

L
⁢
P

-
X
-

(

X
⁢
a

)

m

-
Y
-
Z

(
II
)

wherein
X represents a group represented by the formula (1):

wherein
R 2 is a hydrogen atom or a hydrocarbon group,
Y is a peptide residue consisting of four amino acids and/or analogs thereof and contains a group represented by the formula (2):

wherein
R 1 and B 1 are as defined above, and R 3 represents a hydrogen or a hydrocarbon group,
m number of X a , m and Z are as defined above, and
LP is present or absent and, when present, represents a peptide residue consisting of 1 to 100 amino acids and/or analogs thereof,
and forming the nitrogen-containing 6-membered ring A while eliminating LP, if present, to form the two or more cyclic compounds represented by the formula (I).

2. The production method according to claim 1 , wherein LP is a peptide residue consisting of 11 or more and 100 or less amino acids and/or analogs thereof.

3. The production method according to claim 1 , wherein m is an integer selected from 2 to 24.

4. The production method according to claim 1 , wherein Y is a group represented by the following formula (3):

wherein R 1 , R 3 , R 4 and R 5 each independently represent a hydrogen atom or a hydrocarbon group, and B 1 and B 2 each independently represent an oxygen atom, a sulfur atom or a NH group.

5. The production method according to claim 1 , wherein Y is a group represented by the following formula (3′):

wherein R 1 , R 3 , R 4 and R 5 each independently represent a hydrogen atom or a methyl group, and B 1 and B 2 each independently represent an oxygen atom or a sulfur atom, with the proviso that when B 1 and/or B 2 is a sulfur atom, each of R 1 and R 5 is a hydrogen.

6. The production method according to claim 1 , wherein Y is a group represented by the following formula (3″):

wherein R 1 , R 4 and R 5 each independently represent a hydrogen atom or a methyl group, and B 1 and B 2 each independently represent an oxygen atom or a sulfur atom, with the proviso that when B 1 and/or B 2 is a sulfur atom, each of R 1 and R 5 is a hydrogen.

7. The production method according to claim 1 , wherein Y is a group represented by the following formula (3-1):

8. The production method according to claim 1 , wherein the macrocyclase comprises LazC and/or an enzyme having homology with LazC.

9. The method for producing a compound library according to claim 1 ,

the method further comprising:
producing a mRNA library encoding precursor peptides represented by the formula (III):

LP
′

-

X
′

-

(

Xa
′

)

m
′

-

Y
′

-

Z
′

(
III
)

wherein
X′ is serine or threonine, or an analog thereof,
Y′ is a peptide consisting of four amino acids represented by -Y′(10)-Y′(11)-Y′(12)-Y′(13)-and/or analogs thereof, wherein Y′(10) is an amino acid residue selected from the group consisting of any amino acids and analogs thereof, Y′(11) is serine, cysteine, threonine or diaminopropionic acid, or an analog thereof, Y′(12) is serine or threonine, or an analog thereof, and Y′(13) is an amino acid residue selected from the group consisting of any amino acids and analogs thereof,
m′ number of X a ′, m′ and Z′ are the same as defined as m number of X a , m and Z, respectively, in the formula (I), and
LP′ is present or absent and, when present, represents a peptide consisting of 1 to 100 amino acids and/or analogs thereof;
expressing the precursor peptides by a cell-free translation system with the mRNA library to produce a first peptide library;
forming an azole ring on at least Y′(11) in the precursor peptides with an azole ring-forming enzyme;
converting at least X′ and Y′(12) in the precursor peptides to α,β-unsaturated amino acid residues with an «, β-unsaturated amino acid-forming enzyme in the presence of co-substrate tRNA Glu for glutamylation and aminoacylation enzyme GluRS; and
forming a nitrogen-containing 6-membered ring with a macrocyclase while eliminating LP′, if present.

10. The production method according to claim 9 , wherein
Y′(10) is serine or threonine, o

Assignees

• Univ Tokyo

Inventors

• Suga Hiroaki
• Goto Yuki
• Onaka Hiroyasu
• Vinogradov Alexander

Classifications

• C40B40/10

  Libraries containing peptides or polypeptides, or derivatives thereof · CPC title

• G01N2500/00

  Screening for compounds of potential therapeutic value · CPC title

• G01N33/53

  Immunoassay; Biospecific binding assay; Materials therefor · CPC title

• C12P17/188

  Heterocyclic compound containing in the condensed system at least one hetero ring having nitrogen atoms and oxygen atoms as the only ring heteroatoms (ergot-alcaloids C12P17/183) · CPC title

• C12P17/185

  Heterocyclic compounds containing sulfur atoms as ring hetero atoms in the condensed system (cepam nucleus C12P35/00; penam nucleus C12P37/00) · CPC title