Amino acid active ester and salt thereof

Abstract

The present invention provides novel active esters of amino acids for use in aminoacylation of tRNAs with reduced safety and waste risks.SOLUTION: The present invention provides a compound represented by formula (I): or a salt thereof, in particular dichloropyridinylmethyl ester and 2,2,2 - trifluoroethyl ester or a salt thereof.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present disclosure relates to an amino acid activated ester and a method for producing an aminoacylated tRNA using the ester. [Background technology]

[0002] In recent years, in the field of peptide medicines, non-natural amino acids containing non-natural amino acids are expected to not only exhibit high affinity and specificity for target molecules, but also to have excellent stability and membrane permeability in vivo, making them important candidates for drug discovery. Among the non-natural peptide synthesis methods reported to date, there is a method for synthesizing non-natural peptides from various amino acids and amino acid derivatives, including non-natural amino acids, using flexizyme, which catalyzes the aminoacylation of tRNA (Patent Documents 1 to 6, Non-Patent Documents 4 and 5). In this method, the amino acids used for the aminoacylation of tRNA have not been selected from the conventional methods. Dinitrobenzyl ester (DBE) and cyanomethyl ester (CME) have been used as activated esters. Furthermore, the use of various esters, including picolyl esters, pyridoxy esters, and 2,2,2-trifluoroethyl esters, in amino acid ester synthesis and peptide synthesis has been disclosed (Patent Documents 7 to 13, Non-Patent Documents 1 to 3). Patent Document 14 discloses the use of 2,6-dichloro-4-pyridinemethanol derivatives as pesticides. [Prior art documents] [Patent documents]

[0003] Patent Document 1: WO2007 / 066627(A1) Patent Document 2: WO2008 / 059823(A1) Patent Document 3: WO2011 / 049157(A1) Patent Document 4: WO2015 / 030014(A1) Patent Document 5: Japanese Patent Application Laid-Open No. 2008-125396 Patent document 6 WO2020 / 040840(A2) Patent Document 7 British Patent No. 1212533 Patent Document 8: European Patent Publication No. 0450356 Patent Document 9: WO2010 / 057961(A1) Patent Document 10: WO2013 / 100132(A1) Patent Document 11 WO2016 / 118877(A1) Patent Document 12 WO2018 / 174078(A1) Patent Document 13 WO2018 / 225864(A1) Patent Document 14: WO1999 / 012907(A1) [Non-patent literature]

[0004] Non-patent document 1 JAMaclaren, Aust.J.Chem. 1972, 25, 1293-1299. Non-patent document 2 JAMaclaren, Aust.J.Chem. 1978, 31, 1865-1868. Non-patent document 3 Sklyarov et al., Russian Journal of Bioorganic Chemistry 2000, 26, 245-256. Non-patent document 4 Murakami et al., 2003, Chemistry&Biology, 10, 655-662. Non-patent literature 5. Journal of Bioengineering, 93(12), 744. Summary of the Invention

[0005] Problems that the invention aims to solve DBE and CME, which have traditionally been used as activated esters of amino acids for aminoacylation of tRNA, are explosive nitro compounds and highly toxic cyanide compounds, respectively. Therefore, ensuring safety during use, storage, and disposal requires careful and complicated handling, which in turn requires costs.

[0006] The present invention provides a novel activated ester of an amino acid for use in the aminoacylation of tRNA, which is easy to handle during use, storage, and disposal, and can reduce handling costs. Means to solve the problem

[0007] The inventors conducted extensive research to find an activated ester that could replace DBE or CME, and as a result discovered dichloropyridinylmethyl ester and 2,2,2-trifluoroethyl ester. The use of these activated amino acid esters confirmed that Flexizyme-mediated aminoacylation of tRNA proceeds, and they also found that handling during use, storage, and disposal is easier and handling costs can be reduced.

[0008] This specification includes the disclosure of the following inventions. [1-1] Formula (I):

[0009] [ka]

[0010] [Wherein X represents a pyridyl group substituted with two or more halogen atoms, or a C group substituted with three or more fluorine atoms. 1-3 is alkyl; Q is

[0011] [ka]

[0012] is a group represented by n is an integer of 0 to 3, preferably an integer of 0 to 2, R 7 and R 8 are each independently a hydrogen atom or C 1-3 alkyl; R 1 and R 2 are each independently a hydrogen atom, a halogen atom, cyano, or Y1 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 1 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 1 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 2 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 2 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 2 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 3 C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 3 a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of: 11 , and -CONR 12 R 13 Selected from, or R 1 and R 5 together with the carbon and nitrogen atoms to which they are attached, form Y 2 and forming a 3- to 10-membered heterocycle which may be substituted by one or more substituents selected from R 1 and R 2 together with the carbon atoms to which they are attached, form Y 2 C optionally substituted with one or more substituents selected from 3-10 Carbocycle, or Y 2 and forming a 3- to 10-membered heterocycle which may be substituted by one or more substituents selected from R 3 and R 4 are each independently a hydrogen atom, a halogen atom, cyano, or Y 1 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 1C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 1 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 2 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 2 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 2 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 3 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 3 a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of: 11 , and -CONR 12 R 13 Selected from, or R 3 and R 4 together with the carbon atoms to which they are attached, form Y 2 C optionally substituted with one or more substituents selected from 3-10 Carbocycle, or Y 2 and forming a 3- to 10-membered heterocycle which may be substituted by one or more substituents selected from R 11 , R 12 , and R 13 are each independently, Y 1 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 1 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 1 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 2 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 2C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 2 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 3 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 3 5-14 membered heteroaryl optionally substituted by one or more substituents selected from Y 1 are each independently a halogen atom, nitro, cyano, Y 5 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 5 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 6 C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 6 a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of -SH, -COOR 21 , -NR 22 R 23 , -S(O) p R 24 , -OR 25 , -SO2NR 26 R 27 , and -CONR 28 R 29 and oxo; Y 2 are each independently a halogen atom, nitro, cyano, Y 4 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 4 C optionally substituted with one or more substituents selected from 2-10Alkynyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 5 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 6 C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 6 a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of -SH, -COOR 21 , -NR 22 R 23 , -S(O) p R 24 , -OR 25 , -SO2NR 26 R 27 , and -CONR 28 R 29 and oxo; Y 3 are each independently a halogen atom, nitro, cyano, Y 4 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 5 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 6 C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 6a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of -SH, -COOR 21 , -NR 22 R 23 , -S(O) p R 24 , -OR 25 , -SO2NR 26 R 27 , and -CONR 28 R 29 is selected from R 21 , R 26 , R 27 , R 28 , and R 29 are each independently a hydrogen atom, Y 4 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 5 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 6 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 6 5-14 membered heteroaryl optionally substituted by one or more substituents selected from R 22 , and R 25 are each independently a hydrogen atom, Y 4 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y4 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 5 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 6 C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 6 a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of: a1 , -CONR a2 R a3 , -COOR a4 , -C(=NR a5 )NR a6 R a7 , and -SO2NR a8 R a9 is selected from R 23 is a hydrogen atom, Y 4 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 5 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 6 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 65-14 membered heteroaryl optionally substituted by one or more substituents selected from R 24 is Y 4 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 5 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 6 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 6 5-14 membered heteroaryl optionally substituted by one or more substituents selected from R a1 , R a2 , R a3 , R a4 , R a5 , R a6 , R a7 , R a8 , and R a9 are each independently a hydrogen atom, Y 4 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 5C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 5 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 6 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 6 5-14 membered heteroaryl optionally substituted by one or more substituents selected from Y 4 are each independently a halogen atom, nitro, cyano, Y 5 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 5 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 6 C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 6 a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of -SH, -COOR 31 , -NR 32 R 33 , -S(O) p R 34 , -OR 35 , -SO2NR 36 R 37 , and -CONR 38 R 39 and oxo; Y 5 are each independently a halogen atom, nitro, cyano, Y 7 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 7 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 7 C optionally substituted with one or more substituents selected from 2-10Alkynyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 8 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 9 C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 9 a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of -SH, -COOR 31 , -NR 32 R 33 , -S(O) p R 34 , -OR 35 , -SO2NR 36 R 37 , -CONR 38 R 39 and oxo; Y 6 are each independently a halogen atom, nitro, cyano, Y 7 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 7 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 7 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 8 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 9 C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 9a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of -SH, -COOR 31 , -NR 32 R 33 , -S(O) p R 34 , -OR 35 , -SO2NR 36 R 37 , and -CONR 38 R 39 is selected from R 31 , R 36 , R 37 , R 38 , and R 39 are each independently a hydrogen atom, Y 7 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 7 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 7 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 8 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 9 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 9 5-14 membered heteroaryl optionally substituted by one or more substituents selected from R 32 , and R 35 are each independently a hydrogen atom, Y 7 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 7 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y7 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 8 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 9 C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 9 a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of: b1 , -CONR b2 R b3 , -CO2R b4 , -C(=NR b5 )NR b6 R b7 , and -SO2NR b8 R b9 is selected from R 33 is a hydrogen atom, Y 7 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 7 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 7 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 8 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 9 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 95-14 membered heteroaryl optionally substituted by one or more substituents selected from R 34 is Y 7 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 7 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 7 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 8 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 9 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 9 5-14 membered heteroaryl optionally substituted by one or more substituents selected from R b1 , R b2 , R b3 , R b4 , R b5 , R b6 , R b7 , R b8 , and R b9 are each independently a hydrogen atom, Y 7 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 7 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 7 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 8C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 8 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 9 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 9 5-14 membered heteroaryl optionally substituted by one or more substituents selected from Y 7 are each independently a halogen atom, nitro, cyano, Y 10 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 11 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 12 C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 12 a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of -SH, -COOR 40 , -NR 41 R 42 , -S(O) p R 43 , -OR 44 , -SO2NR 45 R 46 , and -CONR 47 R 48 and oxo; Y 8are each independently a halogen atom, nitro, cyano, Y 10 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 11 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 12 C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 12 a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of -SH, -COOR 40 , -NR 41 R 42 , -S(O) p R 43 , -OR 44 , -SO2NR 45 R 46 , -CONR 47 R 48 and oxo; Y 9 are each independently a halogen atom, nitro, cyano, Y 10 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 11C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 11 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 12 C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 12 a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of -SH, -COOR 40 , -NR 41 R 42 , -S(O) p R 43 , -OR 44 , -SO2NR 45 R 46 , and -CONR 47 R 48 is selected from R 40 , R 45 , R 46 , R 47 , and R 48 are each independently a hydrogen atom, Y 10 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 11 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 12 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 12 5-14 membered heteroaryl optionally substituted by one or more substituents selected from R 41 , and R 44 are each independently a hydrogen atom, Y 10 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 11 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 12 C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 12 a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of: c1 , -CONR c2 R c3 , -COOR c4 , -C(=NR c5 )NR c6 R c7 , and -SO2NR c8 R c9 is selected from R 42 is a hydrogen atom, Y 10 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 11C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 11 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 12 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 12 5-14 membered heteroaryl optionally substituted by one or more substituents selected from R 43 is Y 10 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 11 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 12 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 12 5-14 membered heteroaryl optionally substituted by one or more substituents selected from R c1 , R c2 , R c3 , R c4 , R c5 , R c6 , R c7 , R c8 , and R c9 are each independently a hydrogen atom, Y 10 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 10C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 11 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 12 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 12 5-14 membered heteroaryl optionally substituted by one or more substituents selected from Y 10 each independently represents a C optionally substituted with one or more substituents selected from a halogen atom, nitro, cyano, a halogen atom, and an alkoxy; 1-10 Alkoxy, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 3-10 Cycloalkyl, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 3-10 Cycloalkenyl, halogen atoms and C 1-6 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from alkoxy, a halogen atom, and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 6-14 Aryl, halogen atoms and C 1-6 5-14 membered heteroaryl optionally substituted by one or more substituents selected from alkoxy, a halogen atom, and C 1-6 alkoxy (C 1-10 Alkoxy)carbonyl, halogen atoms and C 1-6Di(C) optionally substituted with one or more substituents selected from alkoxy 1-6 alkyl)aminocarbonyl, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 Alkoxy, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 Alkylthio, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 Alkyl sulfanyl, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 Alkyl sulfinyl, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 alkylsulfonyl, and oxo; Y 11 each independently represents a C optionally substituted by one or more substituents selected from a halogen atom, nitro, cyano, a halogen atom, and an alkoxy; 1-10 C optionally substituted with one or more substituents selected from alkyl, halogen atoms, and alkoxy 2-10 C optionally substituted by one or more substituents selected from alkenyl, halogen atoms, and alkoxy 2-10 C optionally substituted by one or more substituents selected from alkynyl, halogen atoms, and alkoxy 1-10 Alkoxy, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 3-10 Cycloalkyl, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 3-10 Cycloalkenyl, halogen atoms and C 1-6 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from alkoxy, a halogen atom, and C1-6 C optionally substituted with one or more substituents selected from alkoxy 6-14 Aryl, halogen atoms and C 1-6 5-14 membered heteroaryl optionally substituted by one or more substituents selected from alkoxy, a halogen atom, and C 1-6 alkoxy (C 1-10 Alkoxy)carbonyl, halogen atoms and C 1-6 Di(C) optionally substituted with one or more substituents selected from alkoxy 1-6 alkyl)aminocarbonyl, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 Alkoxy, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 Alkylthio, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 Alkyl sulfanyl, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 Alkyl sulfinyl, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 alkylsulfonyl, and oxo; Y 12 each independently represents a C optionally substituted by one or more substituents selected from a halogen atom, nitro, cyano, a halogen atom, and an alkoxy; 1-10 C optionally substituted with one or more substituents selected from alkyl, halogen atoms, and alkoxy 2-10 C optionally substituted by one or more substituents selected from alkenyl, halogen atoms, and alkoxy 2-10 C optionally substituted by one or more substituents selected from alkynyl, halogen atoms, and alkoxy 1-10Alkoxy, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 3-10 Cycloalkyl, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 3-10 Cycloalkenyl, halogen atoms and C 1-6 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from alkoxy, a halogen atom, and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 6-14 Aryl, halogen atoms and C 1-6 5-14 membered heteroaryl optionally substituted by one or more substituents selected from alkoxy, a halogen atom, and C 1-6 alkoxy (C 1-10 Alkoxy)carbonyl, halogen atoms and C 1-6 Di(C) optionally substituted with one or more substituents selected from alkoxy 1-6 alkyl)aminocarbonyl, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 Alkoxy, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 Alkylthio, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 Alkyl sulfanyl, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 Alkyl sulfinyl, and halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 alkylsulfonyl; p's are each independently an integer of 0 to 2, R 5is a hydrogen atom, and Y is on the benzene ring. 13 benzyl optionally substituted with one or more substituents selected from the group consisting of 13 benzyloxycarbonyl optionally substituted by one or more substituents selected from 13 phenylcarbonyl optionally substituted by one or more substituents selected from (C 1-10 alkoxy)carbonyl, (C 1-10 alkyl)carbonyl, Y 1 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 1 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 1 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 2 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 2 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 2 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 3 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 3 5-14 membered heteroaryl optionally substituted by one or more substituents selected from R 6 is a hydrogen atom, and Y is on the benzene ring. 13 benzyl optionally substituted with one or more substituents selected from the group consisting of 13 benzyloxycarbonyl optionally substituted by one or more substituents selected from 13 phenylcarbonyl optionally substituted by one or more substituents selected from (C 1-10 alkoxy)carbonyl, (C 1-10 alkyl)carbonyl, or R 5and R 6 form a 5- to 14-membered nitrogen-containing heterocycle having an imide structure together with the nitrogen atom to which they are bonded, Y 13 are each independently a halogen atom, C 1-10 Alkoxy, and C 1-10 alkyl] or a salt thereof.

[0013] [1-2] The compound according to [1-1], wherein n is 0 or 1, or a salt thereof. [1-3] R 1 The compound according to [1-1] or [1-2], or a salt thereof, wherein [1-4] R 7 and R 8 The compound or salt thereof according to any one of [1-1] to [1-3], wherein is a hydrogen atom.

[0014] [1-5] The compound or salt thereof according to any one of [1-1] to [1-4], wherein X is 2,6-dihalo-4-pyridyl. [1-6] The compound or salt thereof according to any one of [1-1] to [1-4], wherein X is 2,6-dichloro-4-pyridyl.

[0015] [1-7] X is perfluoro C 1-3 The compound according to any one of [1-1] to [1-4], or a salt thereof, wherein R is alkyl. [1-8] The compound or salt thereof according to any one of [1-1] to [1-4], wherein X is trifluoromethyl.

[0016] [1-9] A composition for use in acylation of tRNA, comprising the compound according to any one of [1-1] to [1-8] or a salt thereof. [1-10] The composition according to [1-9], which is used for acylation of tRNA in the presence of Flexizyme.

[0017] [1-11] Formula (I):

[0018] [ka]

[0019] [In the formula, R 1 , R 2 , R 5 , R 6 , R 7 , R 8 , Q and X are as defined in any one of [1-1] to [1-8]. A method for producing a compound represented by formula (II):

[0020] [ka]

[0021] with a compound represented by formula (III):

[0022] [ka]

[0023] wherein L is a leaving group. The method for producing the compound according to the present invention comprises reacting the compound represented by the formula (I) with a compound represented by the formula (I). [1-12] The method according to [1-11], wherein the reaction is carried out in the presence of a base.

[0024] [1-13] The method according to [1-12], wherein the base is diisopropylethylamine. [1-14] A method for producing a tRNA acylated at the 3' end, wherein the acyl group at the 3' end is:

[0025] [ka]

[0026] [In the formula, R 1 , R 2 , R 5 , R 6, and Q is as defined in any of [1-1] to [1-8]. is a group represented by Formula (I):

[0027] [ka]

[0028] [In the formula, R 1 , R 2 , R 5 , R 6 , R 7 , R 8 , Q and X are as defined in any one of [1-1] to [1-8]. The method for producing a tRNA-binding protein as described above, comprising reacting a compound represented by the formula:

[0029] [1-15] A method for preparing a peptide library, comprising: Producing a tRNA acylated at the 3' end by the method described in [1-14]. preparing a library of mRNA; and A peptide library is prepared by synthesizing peptides corresponding to each mRNA from an mRNA library using a cell-free translation system. The method comprising:

[0030] [1-16] A method for acylating the 3' end of tRNA, comprising: converting tRNA into a compound of formula (I):

[0031] [ka]

[0032] [In the formula, R 1 , R 2 , R 5 , R 6 , R 7 , R 8 , Q and X are as defined in any one of [1-1] to [1-8]. In the presence of flexizyme, a compound represented by the formula:

[0033] [ka]

[0034] [In the formula, R 1 , R 2 , R 5 , R 6 , and Q is as defined in any of [1-1] to [1-8]. A method of adding an acyl group represented by the following formula:

[0035] [2-1] Formula (Ia):

[0036] [ka]

[0037] [Wherein X represents a pyridyl group substituted with two or more halogen atoms, or a C group substituted with three or more fluorine atoms. 1-3 is alkyl; Z is -NR 5 R 6 -OR d and Q is

[0038] [ka]

[0039] is a group represented by n is an integer of 0 to 3, preferably an integer of 0 to 2, R 7 and R 8 are each independently a hydrogen atom or C 1-3 alkyl; R 1 and R 2are each independently a hydrogen atom, a halogen atom, cyano, or Y 1 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 1 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 1 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 2 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 2 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 2 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 3 C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 3 a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of: 11 , and -CONR 12 R 13 Selected from, or R 1 and R 5 together with the carbon and nitrogen atoms to which they are attached, form Y 2 and forming a 3- to 10-membered non-aromatic heterocycle which may be substituted by one or more substituents selected from When n is an integer from 1 to 3, R 1 and any one of R 3 together with the carbon atoms to which they are attached, form Y 2 C optionally substituted with one or more substituents selected from 3-10 Carbocycle, or Y 2 and forming a 3- to 10-membered non-aromatic heterocycle which may be substituted by one or more substituents selected from R 1 and R 2 together with the carbon atoms to which they are attached, form Y 2C optionally substituted with one or more substituents selected from 3-10 Carbocycle, or Y 2 and forming a 3- to 10-membered non-aromatic heterocycle which may be substituted by one or more substituents selected from R 3 and R 4 are each independently a hydrogen atom, a halogen atom, cyano, or Y 1 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 1 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 1 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 2 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 2 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 2 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 3 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 3 a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of: 11 , and -CONR 12 R 13 Selected from, or R 3 and R 4 together with the carbon atoms to which they are attached, form Y 2 C optionally substituted with one or more substituents selected from 3-10 Carbocycle, or Y 2 and forming a 3- to 10-membered non-aromatic heterocycle which may be substituted by one or more substituents selected from When n is an integer from 1 to 3, R 5 and any one of R 3together with the nitrogen and carbon atoms to which they are attached, form Y 2 and forming a 3- to 10-membered nitrogen-containing non-aromatic heterocycle which may be substituted by one or more substituents selected from R 11 , R 12 , and R 13 are each independently, Y 1 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 1 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 1 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 2 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 2 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 2 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 3 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 3 5-14 membered heteroaryl optionally substituted by one or more substituents selected from Y 1 are each independently a halogen atom, nitro, cyano, Y 5 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 5 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 6 C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 6a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of -SH, -COOR 21 , -NR 22 R 23 , -N3, -S(O) p R 24 , -OR 25 , -SO2NR 26 R 27 , and -CONR 28 R 29 and oxo; Y 2 are each independently a halogen atom, nitro, cyano, Y 4 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 5 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 6 C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 6 a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of -SH, -COOR 21 , -NR 22 R 23 , -S(O) p R 24 , -OR 25 , -SO2NR 26 R 27 , and -CONR 28 R 29 and oxo; Y 3are each independently a halogen atom, nitro, cyano, Y 4 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 5 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 6 C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 6 a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of -SH, -COOR 21 , -NR 22 R 23 , -S(O) p R 24 , -OR 25 , -SO2NR 26 R 27 , and -CONR 28 R 29 is selected from R 21 , R 26 , R 27 , R 28 , and R 29 are each independently a hydrogen atom, Y 4 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 5C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 5 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 6 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 6 5-14 membered heteroaryl optionally substituted by one or more substituents selected from R 22 , and R 25 are each independently a hydrogen atom, Y 4 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 5 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 6 C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 6 a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of: a1 , -CONR a2 R a3 , -COOR a4 , -C(=NR a5 )NR a6 R a7 , -S(O) p R 24 , and -SO2NRa8 R a9 is selected from R 23 is a hydrogen atom, Y 4 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 5 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 6 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 6 5-14 membered heteroaryl optionally substituted by one or more substituents selected from R 24 is Y 4 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 5 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 6 C optionally substituted with one or more substituents selected from6-14 Aryl, and Y 6 5-14 membered heteroaryl optionally substituted by one or more substituents selected from R a1 , R a2 , R a3 , R a4 , R a5 , R a6 , R a7 , R a8 , and R a9 are each independently a hydrogen atom, Y 4 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 4 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 5 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 6 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 6 5-14 membered heteroaryl optionally substituted by one or more substituents selected from Y 4 are each independently a halogen atom, nitro, cyano, Y 5 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 5 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 5 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 6C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 6 a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of -SH, -COOR 31 , -NR 32 R 33 , -S(O) p R 34 , -OR 35 , -SO2NR 36 R 37 , and -CONR 38 R 39 and oxo; Y 5 are each independently a halogen atom, nitro, cyano, Y 7 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 7 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 7 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 8 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 9 C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 9 a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of -SH, -COOR 31 , -NR 32 R 33 , -S(O) p R 34 , -OR 35 , -SO2NR 36 R 37 , -CONR 38 R 39and oxo; Y 6 are each independently a halogen atom, nitro, cyano, Y 7 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 7 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 7 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 8 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 9 C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 9 a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of -SH, -COOR 31 , -NR 32 R 33 , -S(O) p R 34 , -OR 35 , -SO2NR 36 R 37 , and -CONR 38 R 39 is selected from R 31 , R 36 , R 37 , R 38 , and R 39 are each independently a hydrogen atom, Y 7 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 7 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 7 C optionally substituted with one or more substituents selected from2-10 Alkynyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 8 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 9 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 9 5-14 membered heteroaryl optionally substituted by one or more substituents selected from R 32 , and R 35 are each independently a hydrogen atom, Y 7 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 7 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 7 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 8 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 9 C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 9 a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of: b1 , -CONR b2 R b3 , -CO2R b4 , -C(=NR b5 )NR b6 R b7 , and -SO2NRb8 R b9 is selected from R 33 is a hydrogen atom, Y 7 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 7 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 7 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 8 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 9 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 9 5-14 membered heteroaryl optionally substituted by one or more substituents selected from R 34 is Y 7 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 7 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 7 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 8 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 9 C optionally substituted with one or more substituents selected from6-14 Aryl, and Y 9 5-14 membered heteroaryl optionally substituted by one or more substituents selected from R b1 , R b2 , R b3 , R b4 , R b5 , R b6 , R b7 , R b8 , and R b9 are each independently a hydrogen atom, Y 7 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 7 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 7 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 8 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 8 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 9 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 9 5-14 membered heteroaryl optionally substituted by one or more substituents selected from Y 7 are each independently a halogen atom, nitro, cyano, Y 10 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 11C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 11 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 12 C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 12 a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of -SH, -COOR 40 , -NR 41 R 42 , -S(O) p R 43 , -OR 44 , -SO2NR 45 R 46 , and -CONR 47 R 48 and oxo; Y 8 are each independently a halogen atom, nitro, cyano, Y 10 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 11 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 12 C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 12a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of -SH, -COOR 40 , -NR 41 R 42 , -S(O) p R 43 , -OR 44 , -SO2NR 45 R 46 , -CONR 47 R 48 and oxo; Y 9 are each independently a halogen atom, nitro, cyano, Y 10 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 11 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 12 C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 12 a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of -SH, -COOR 40 , -NR 41 R 42 , -S(O) p R 43 , -OR 44 , -SO2NR 45 R 46 , and -CONR 47 R 48 is selected from R 40 , R 45 , R 46 , R47 , and R 48 are each independently a hydrogen atom, Y 10 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 11 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 12 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 12 5-14 membered heteroaryl optionally substituted by one or more substituents selected from R 41 , and R 44 are each independently a hydrogen atom, Y 10 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 11 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 12C optionally substituted with one or more substituents selected from 6-14 Aryl, Y 12 a 5- to 14-membered heteroaryl optionally substituted by one or more substituents selected from the group consisting of: c1 , -CONR c2 R c3 , -COOR c4 , -C(=NR c5 )NR c6 R c7 , and -SO2NR c8 R c9 is selected from R 42 is a hydrogen atom, Y 10 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 11 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 12 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 12 5-14 membered heteroaryl optionally substituted by one or more substituents selected from R 43 is Y 10 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y11 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 11 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 12 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 12 5-14 membered heteroaryl optionally substituted by one or more substituents selected from R c1 , R c2 , R c3 , R c4 , R c5 , R c6 , R c7 , R c8 , and R c9 are each independently a hydrogen atom, Y 10 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 10 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 11 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 11 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 12 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 12 5-14 membered heteroaryl optionally substituted by one or more substituents selected from Y 10each independently represents a C optionally substituted by one or more substituents selected from a halogen atom, nitro, cyano, a halogen atom, and an alkoxy; 1-10 Alkoxy, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 3-10 Cycloalkyl, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 3-10 Cycloalkenyl, halogen atoms and C 1-6 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from alkoxy, a halogen atom, and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 6-14 Aryl, halogen atoms and C 1-6 5-14 membered heteroaryl optionally substituted by one or more substituents selected from alkoxy, a halogen atom, and C 1-6 alkoxy (C 1-10 Alkoxy)carbonyl, halogen atoms and C 1-6 Di(C) optionally substituted with one or more substituents selected from alkoxy 1-6 alkyl)aminocarbonyl, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 Alkoxy, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 Alkylthio, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 Alkyl sulfanyl, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 Alkyl sulfinyl, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6alkylsulfonyl, and oxo; Y 11 each independently represents a C optionally substituted by one or more substituents selected from a halogen atom, nitro, cyano, a halogen atom, and an alkoxy; 1-10 C optionally substituted with one or more substituents selected from alkyl, halogen atoms, and alkoxy 2-10 C optionally substituted by one or more substituents selected from alkenyl, halogen atoms, and alkoxy 2-10 C optionally substituted by one or more substituents selected from alkynyl, halogen atoms, and alkoxy 1-10 Alkoxy, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 3-10 Cycloalkyl, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 3-10 Cycloalkenyl, halogen atoms and C 1-6 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from alkoxy, a halogen atom, and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 6-14 Aryl, halogen atoms and C 1-6 5-14 membered heteroaryl optionally substituted by one or more substituents selected from alkoxy, a halogen atom, and C 1-6 alkoxy (C 1-10 Alkoxy)carbonyl, halogen atoms and C 1-6 Di(C) optionally substituted with one or more substituents selected from alkoxy 1-6 alkyl)aminocarbonyl, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 Alkoxy, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6Alkylthio, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 Alkyl sulfanyl, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 Alkyl sulfinyl, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 alkylsulfonyl, and oxo; Y 12 each independently represents a C optionally substituted by one or more substituents selected from a halogen atom, nitro, cyano, a halogen atom, and an alkoxy; 1-10 C optionally substituted with one or more substituents selected from alkyl, halogen atoms, and alkoxy 2-10 C optionally substituted by one or more substituents selected from alkenyl, halogen atoms, and alkoxy 2-10 C optionally substituted by one or more substituents selected from alkynyl, halogen atoms, and alkoxy 1-10 Alkoxy, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 3-10 Cycloalkyl, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 3-10 Cycloalkenyl, halogen atoms and C 1-6 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from alkoxy, a halogen atom, and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 6-14 Aryl, halogen atoms and C 1-6 5-14 membered heteroaryl optionally substituted by one or more substituents selected from alkoxy, a halogen atom, and C 1-6 alkoxy (C 1-10Alkoxy)carbonyl, halogen atoms and C 1-6 Di(C) optionally substituted with one or more substituents selected from alkoxy 1-6 alkyl)aminocarbonyl, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 Alkoxy, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 Alkylthio, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 Alkyl sulfanyl, halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 Alkyl sulfinyl, and halogen atoms and C 1-6 C optionally substituted with one or more substituents selected from alkoxy 1-6 alkylsulfonyl; p's are each independently an integer of 0 to 2, R 5 is a hydrogen atom, and Y is on the benzene ring. 13 benzyl optionally substituted with one or more substituents selected from the group consisting of 13 benzyloxycarbonyl optionally substituted by one or more substituents selected from 13 phenylcarbonyl optionally substituted by one or more substituents selected from (C 1-10 alkoxy)carbonyl, optionally substituted by halogen atoms (C 1-10 alkyl)carbonyl, Y 1 C optionally substituted with one or more substituents selected from 1-10 Alkyl, Y 1 C optionally substituted with one or more substituents selected from 2-10 Alkenyl, Y 1 C optionally substituted with one or more substituents selected from 2-10 Alkynyl, Y 2C optionally substituted with one or more substituents selected from 3-10 Cycloalkyl, Y 2 C optionally substituted with one or more substituents selected from 3-10 cycloalkenyl, Y 2 a 3- to 14-membered non-aromatic heterocyclyl optionally substituted by one or more substituents selected from the group consisting of 3 C optionally substituted with one or more substituents selected from 6-14 Aryl, and Y 3 5-14 membered heteroaryl optionally substituted by one or more substituents selected from R 6 is a hydrogen atom, and Y is on the benzene ring. 13 benzyl optionally substituted with one or more substituents selected from the group consisting of 13 benzyloxycarbonyl optionally substituted by one or more substituents selected from 13 phenylcarbonyl optionally substituted by one or more substituents selected from (C 1-10 alkoxy)carbonyl, (C 1-10 alkyl)carbonyl, or R 5 and R 6 are taken together with the nitrogen atom to which they are attached to form a 5- to 14-membered nitrogen-containing heterocyclyl having an imido structure, or Y 2 and forming a 3- to 14-membered non-aromatic heterocyclyl containing two or more nitrogen atoms as ring atoms which may be substituted by one or more substituents selected from R d is a hydrogen atom, and Y is on the benzene ring. 13 benzyl optionally substituted with one or more substituents selected from the group consisting of 13 benzyloxycarbonyl optionally substituted by one or more substituents selected from 13 phenylcarbonyl optionally substituted by one or more substituents selected from (C 1-10 alkoxy)carbonyl, (C 1-10 alkyl)carbonyl; Y13 are each independently a halogen atom, C 1-10 Alkoxy, and C 1-10 alkyl, When the 3- to 14-membered non-aromatic heterocyclyl is a monocyclic heterocyclyl, the heterocyclyl may be fused with a benzene ring, Said C 6-14 When the aryl is phenyl, the phenyl may be fused with a 5- to 7-membered non-aromatic heterocycle. or a salt thereof (wherein Z is hydroxy, n is 0, and R 7 and R 8 is a hydrogen atom, X is 2,6-dichloropyridin-4-yl, R 1 and R 2 is a hydrogen atom, or R 1 is a hydrogen atom, and R 2 is methyl, ethyl, isopropyl, n-butyl, tert-butyl, or isobutyl, or R 1 is ethyl and R 2 (Excluding compounds where is methyl or ethyl).

[0040] [2-2] The compound according to [2-1], wherein n is 0 or 1, or a salt thereof. [2-3] R 1 The compound or salt thereof according to [2-1] or [2-2], wherein [2-4] R 7 and R 8 The compound or salt thereof according to any one of [2-1] to [2-3], wherein is a hydrogen atom.

[0041] [2-5] The compound or salt thereof according to any one of [2-1] to [2-4], wherein X is 2,6-dihalo-4-pyridyl. [2-6] The compound or salt thereof according to any one of [2-1] to [2-4], wherein X is 2,6-dichloro-4-pyridyl.

[0042] [2-7] X is perfluoro C 1-3 The compound according to any one of [2-1] to [2-4], or a salt thereof, wherein R is alkyl. [2-8] The compound or salt thereof according to any one of [2-1] to [2-4], wherein X is trifluoromethyl.

[0043] [2-9] A composition for use in acylation of tRNA, comprising the compound according to any one of [2-1] to [2-8] or a salt thereof. [2-10] The composition according to [2-9], which is used for acylation of tRNA in the presence of flexizyme.

[0044] [2-11] Formula (Ia):

[0045] [ka]

[0046] [In the formula, R 1 , R 2 , R 7 , R 8 , Q, X and Z are as defined in any of [2-1] to [2-8]. A method for producing a compound represented by formula (IIa):

[0047] [ka]

[0048] with a compound represented by formula (III):

[0049] [ka]

[0050] wherein L is a leaving group. The method for producing the compound according to the present invention comprises reacting the compound represented by the formula (I) with a compound represented by the formula (I). [2-12] The method according to [2-11], wherein the reaction is carried out in the presence of a base.

[0051] [2-13] The method according to [2-12], wherein the base is diisopropylethylamine. [2-14] A method for producing a tRNA acylated at the 3' end, wherein the acyl group at the 3' end is:

[0052] [ka]

[0053] [In the formula, R 1 , R 2 , Q and Z are as defined in any one of [2-1] to [2-8]. is a group represented by Formula (Ia):

[0054] [ka]

[0055] [In the formula, R 1 , R 2 , R 7 , R 8 , Q, X and Z are as defined in any of [2-1] to [2-8]. The method for producing a tRNA-binding protein as described above, comprising reacting a compound represented by the formula:

[0056] [2-15] A method for preparing a peptide library, comprising: Producing a tRNA acylated at the 3' end by the method described in [2-14]. preparing a library of mRNA; and A peptide library is prepared by synthesizing peptides corresponding to each mRNA from an mRNA library using a cell-free translation system. The method comprising:

[0057] [2-16] A method for acylating the 3' end of tRNA, comprising: converting tRNA into a compound of formula (I):

[0058] [ka]

[0059] [In the formula, R 1 , R 2 , R 7 , R 8 , Q, X and Z are as defined in any of [2-1] to [2-8]. In the presence of flexizyme, a compound represented by the formula:

[0060] [ka]

[0061] [In the formula, R 1 , R 2 , R 5 , R 6 , and Q is as defined in any one of [2-1] to [2-8]. A method of adding an acyl group represented by the following formula: [2-17] Z is -NR 5 R 6 The compound according to [2-1], or a salt thereof, Effect of the invention

[0062] The amino acid activated esters of the present invention are useful as synthetic intermediates, for example, as reagents for acylation reactions. In one aspect of the present invention, the activated esters are used in the aminoacylation of tRNA by flexizyme, and are useful for synthesizing unique peptides composed of various amino acids and amino acid derivatives, including unnatural amino acids. Furthermore, the novel amino acid activated esters of the present invention are easy to handle during use, storage, and disposal, and can also reduce handling costs. [Brief explanation of the drawings]

[0063] [Figure 1-1] FIG. 1-1 is a photograph, instead of a drawing, showing the results of confirming the efficiency of acylation of (2,6-dichloropyridin-4-yl)methyl L-isoleucinate (Compound No. 21). [Figure 1-2] Figure 1-2 is a photograph, instead of a drawing, showing the results of confirming the efficiency of acylation of (2,6-dichloropyridin-4-yl)methyl glycinate (compound number 99). [Figure 1-3] Figure 1-3 is a photograph, instead of a drawing, showing the results of confirming the efficiency of acylation of 2,2,2-trifluoroethyl (S)-2-amino-4-phenylbutanoate (Compound No. 47). [Figure 1-4] Figure 1-4 is a photograph, instead of a drawing, showing the results of confirming the efficiency of acylation of 2,2,2-trifluoroethyl (S)-2-amino-3-(1H-pyrrolo[2,3-b]pyridin-3-yl)propanoate (compound number 81). [Figure 2] FIG. 2 is a graph showing the results of confirming the risk of each activated ester of amino acid, using 2,4-dinitrotoluene (compound number 115) and benzoyl peroxide (compound number 116) as reference compounds, plotting the reference points of QDSC and TDSC, and connecting the two points to form a risk judgment line.

[0064] As used herein, "halogen atom" refers to a fluorine atom, a chlorine atom, a bromine atom, an iodine atom, or the like. When a halogen atom is used as a substituent for aryl, heteroaryl, or the like, preferred examples include a fluorine atom, a chlorine atom, and a bromine atom. When a halogen atom is used as a substituent for alkyl or a group containing alkyl as a part thereof (alkoxy, alkenyl, alkylthio, or the like) as used herein, preferred examples include a fluorine atom. Specific examples of groups having a halogen atom as a substituent include trifluoromethyl, pentafluoroethyl, trifluoromethoxy, pentafluoroethoxy, trifluoromethylthio, and pentafluoroethylthio.

[0065] As used herein, "C 1-3 "Alkyl" means a monovalent group derived by removing any one hydrogen atom from a straight-chain or branched-chain saturated aliphatic hydrocarbon having 1 to 3 carbon atoms. Specific examples include methyl, ethyl, n-propyl, and isopropyl.

[0066] As used herein, "C 1-10 The term "alkyl" refers to a monovalent group derived by removing any one hydrogen atom from a linear or branched saturated aliphatic hydrocarbon having 1 to 10 carbon atoms. Specific examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, 1-methylpropyl, n-pentyl, isopentyl, 2-methylbutyl, 1,1-dimethylpropyl, 1-ethylpropyl, hexyl, 4-methylpentyl, n-heptyl, 5-methylhexyl, 1-propylbutyl, 2-ethyl-2-methylbutyl, n-octyl, 5-methylheptyl, 2,3-dimethylhexyl, 1-methyl-1-propylbutyl, 2,2-diethylbutyl, 7-methyloctyl, 5-ethylheptyl, n-decyl, 8-methylnonyl, 5,5-dimethyloctyl, and 4-ethyl-6-methylheptyl.

[0067] As used herein, "C 2-10 "Alkenyl" refers to a monovalent group derived by removing any one hydrogen atom from a straight-chain or branched-chain aliphatic hydrocarbon having 2 to 10 carbon atoms and at least one double bond (two adjacent SP2 carbon atoms). Specific examples include vinyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-methyl-2-propenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-methyl-2-butenyl, 1-methyl-3-butenyl, 2-methyl-3-butenyl, 3-methyl-1-butenyl, 1,1-dimethyl-2-propenyl, 1-hexenyl, heptenyl, and octenyl.

[0068] As used herein, "optionally substituted C2-10 "Alkenyl" refers to an unsubstituted C 2-10 Alkenyl or C in which one or more hydrogen atoms on the alkenyl are replaced by a given substituent 2-10 It means alkenyl. When it has two or more substituents, the substituents may be the same or different. One carbon atom may be substituted with multiple substituents.

[0069] As used herein, "C 2-10 "Alkynyl" refers to a monovalent group derived by removing any one hydrogen atom from a straight-chain or branched-chain aliphatic hydrocarbon having 2 to 10 carbon atoms and at least one triple bond (two adjacent SP carbon atoms). Specific examples include ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-methyl-2-propynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 1-methyl-2-butynyl, 1-methyl-3-butynyl, 2-methyl-3-butynyl, 3-methyl-1-butynyl, 1,1-dimethyl-2-propynyl, 1-hexynyl, heptynyl, heptadiynyl, octynyl, and octadiynyl.

[0070] As used herein, "optionally substituted C 2-10 "Alkynyl" refers to an unsubstituted C 2-10 Alkynyl or C in which one or more hydrogen atoms on the alkynyl are replaced by a given substituent 2-10 It means alkynyl. When it has two or more substituents, the substituents may be the same or different. One carbon atom may be substituted with multiple substituents.

[0071] As used herein, "C 1-6 "Alkoxy" is C 1-6 means an alkyl-O- group, where C 1-6 Alkyl is as defined above. Specific examples include methoxy, ethoxy, 1-propoxy, 2-propoxy, n-butoxy, i-butoxy, sec-butoxy, t-butoxy, 1-pentyloxy, and 1-hexyloxy.

[0072] As used herein, "C 1-10 "Alkoxy" is C 1-10 means an alkyl-O- group, where C 1-10 The alkyl is as defined above. Specific examples include methoxy, ethoxy, 1-propoxy, 2-propoxy, n-butoxy, i-butoxy, sec-butoxy, t-butoxy, 1-pentyloxy, 1-hexyloxy, n-heptyloxy, 5-methylhexyloxy, 1-propylbutyloxy, 2-ethyl-2-methylbutyloxy, n-octyloxy, 5-methylheptyloxy, 2,3-dimethylhexyloxy, 1-methyl-1-propylbutyloxy, and 2,2-diethylbutyloxy, 7-methyloctyloxy, 5-ethylheptyloxy, n-decyloxy, 8-methylnonyloxy, 5,5-dimethyloctyloxy, and 4-ethyl-6-methylheptyloxy.

[0073] As used herein, "C 1-6 "Alkylthio" means C 1-6 means an alkyl-S- group, where C 1-6 Alkyl is as defined above. Specific examples include methylthio, ethylthio, n-propylthio, i-propylthio, n-butylthio, i-butylthio, t-butylthio, sec-butylthio, 1-methylpropylthio, n-pentylthio, isopentylthio, 2-methylbutylthio, 1,1-dimethylpropylthio, 1-ethylpropylthio, hexylthio, 4-methylpentylthio, and 2-ethylbutylthio.

[0074] In this specification, "(C 1-10 "Alkyl)carbonyl" is C 1-10 alkyl-C(O)- group, where C 1-10The alkyl is as defined above. Specific examples include methylcarbonyl, ethylcarbonyl, n-propylcarbonyl, i-propylcarbonyl, n-butylcarbonyl, i-butylcarbonyl, sec-butylcarbonyl, t-butylcarbonyl, 1-methylpropylcarbonyl, n-pentylcarbonyl, isopentylcarbonyl, 2-methylbutylcarbonyl, 1,1-dimethylpropylcarbonyl, 1-ethylpropylcarbonyl, hexylcarbonyl, 4-methylpentylcarbonyl, and 2-ethylbutylcarbonyl.

[0075] As used herein, "(di(C 1-6 alkyl)aminocarbonyl" is a di(C 1-6 a "Di(C 1-6 "Alkyl)amino" is a compound consisting of two C 1-6 It means an amino substituted with alkyl. Specific examples include dimethylaminocarbonyl, diethylaminocarbonyl, and the like.

[0076] As used herein, "C 1-6 "Alkylsulfanyl" is C 1-6 means an alkyl-S- group, where C 1-6 The alkyl is as defined above. Specific examples include methylsulfanyl, ethylsulfanyl, n-propylsulfanyl, etc., and preferred is methylsulfanyl.

[0077] As used herein, "C 1-6 "Alkylsulfonyl" is C 1-6 means an alkyl-SO2- group, where C 1-6 The alkyl is as defined above. Specific examples include methylsulfonyl, ethylsulfonyl, n-propylsulfonyl, etc., and preferred is methylsulfonyl.

[0078] As used herein, "C 1-6 "Alkylsulfinyl" is C 1-6 means an alkyl-S(=O)- group, where C1-6 The alkyl is as defined above. Specific examples include methylsulfinyl, ethylsulfinyl, n-propylsulfinyl, etc., and preferred is methylsulfinyl.

[0079] In this specification, "(C 1-10 "Alkoxy)carbonyl" is C 1-10 means an alkyl-OC(O)- group, where C 1-10 The alkyl is as defined above. Specific examples include methoxycarbonyl, ethoxycarbonyl, n-propoxycarbonyl, i-propoxycarbonyl, n-butoxycarbonyl, i-butoxycarbonyl, sec-butoxycarbonyl, t-butoxycarbonyl, 1-methylpropoxycarbonyl, n-pentyloxycarbonyl, isopentyloxycarbonyl, 2-methylbutoxycarbonyl, 1,1-dimethylpropoxycarbonyl, 1-ethylpropoxycarbonyl, hexyloxycarbonyl, 4-methylpentyloxycarbonyl, and 2-ethylbutoxycarbonyl.

[0080] As used herein, "C 6-14 "Aryl" means a monovalent aromatic hydrocarbon ring group. 6-14 Aryl includes, for example, phenyl, 1-naphthyl, and 2-naphthyl. C 6-14 When the aryl is phenyl, the phenyl may be fused with a 5- to 7-membered non-aromatic heterocycle, and the C 6-14 An example of an aryl is 2,3-dihydrobenzo-1,4-dioxinyl.

[0081] As used herein, "5- to 14-membered heteroaryl" refers to an aromatic ring group containing one or more (e.g., 1 to 5, preferably 1 to 3) heteroatoms among 5 to 14 ring-constituting atoms. The ring may be a monocyclic or bicyclic ring. Specific examples of "5- to 14-membered heteroaryl" include thienyl, pyridazinyl, pyrazinyl, thiazolyl, oxazolyl, isothiazolyl, thiadiazolyl, oxadiazolyl, isoxazolyl, pyrazolyl, quinolinyl, isoquinolyl, benzothiazolyl, benzoxazolyl, benzimidazolyl, pyridinyl, pyrimidinyl, indolyl, imidazolyl, furyl, thioxazolyl, pyrrolyl, tetrazolyl, oxopyrimidinyl, naphthyl, benzodioxinyl, benzisoxazolyl, benzisothiazolyl, indazolyl, benzothienyl, benzofuranyl, benzopyranyl, and triazolyl.

[0082] As used herein, "3- to 14-membered non-aromatic heterocyclyl" refers to a non-aromatic ring or ring system containing at least one heteroatom among 3 to 14 ring-constituting atoms. The heterocyclyl may have any degree of saturation, as long as at least one ring in the ring system is aromatic. The heteroatom may be present in a non-aromatic or aromatic ring in the ring system. In preferred 6-membered monocyclic heterocyclyls, the heteroatoms are selected from up to three O, N, or S atoms, and in preferred 5-membered monocyclic heterocyclyls, the heteroatoms are selected from up to two O, N, or S atoms. Examples of heterocyclyl include azepinyl, acridinyl, carbazolyl, cinnolinyl, dioxolanyl, imidazolinyl, imidazolidinyl, morpholinyl, oxiranyl, oxepanyl, thiepanyl, pyridyl, piperidinyl, piperazinyl, dioxopiperazinyl, pyrrolidinyl, pyrrolidinoyl, 4-piperidonyl, pyrazolinyl, pyrazolidinyl, oxazolinyl, oxazolidinyl, oxazolidinonyl, thienyl, thiazolinyl, thiazolidinyl, and the like. When the 3- to 14-membered non-aromatic heterocyclyl is a monocyclic heterocyclyl, the heterocyclyl may be fused with a benzene ring, and an example of the 3- to 14-membered non-aromatic heterocyclyl fused with a benzene ring includes at least 2,3-dihydrobenzo-1,4-dioxinyl.

[0083] As used herein, "C 3-10 The "carbocycle" refers to a cycloalkane ring, cycloalkene ring, or cycloalkyne ring having 3 to 10 carbon atoms constituting the ring, and includes, for example, cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, cyclononane, cyclodecane, cyclopropene, cyclobutene, cyclopentene, cyclohexene, cycloheptene, cyclooctene, cyclononene, cyclodecene, cyclohexadiene, cyclooctadiene, and cyclooctyne.

[0084] As used herein, the term "3- to 10-membered heterocycle" refers to a heterocyclic group containing one N heteroatom and consisting of 3 to 10 ring-constituting atoms. Specific examples include pyrrolidine, piperidine, azepane, azocane, etc., and particularly pyrrolidine and piperidine.

[0085] As used herein, "C 3-10 "Cycloalkyl" means a cyclic saturated aliphatic hydrocarbon group having 3 to 10 carbon atoms. Specific examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, and the like.

[0086] As used herein, "C 3-6 "Cycloalkenyl" means a cyclic aliphatic hydrocarbon group having 3 to 6 carbon atoms and at least one double bond (two adjacent SP2 carbon atoms). Specific examples include cyclopropenyl, cyclobutenyl, cyclopentenyl, and cyclohexenyl.

[0087] As used herein, "C 3-10"Cycloalkenyl" means a cyclic aliphatic hydrocarbon group having 3 to 10 carbon atoms and at least one double bond (two adjacent SP2 carbon atoms). Specific examples include cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, cyclononenyl, cyclodekenyl, etc.

[0088] In this specification, the term "5- to 14-membered nitrogen-containing heterocycle having an imide structure" refers to a heterocyclic group consisting of 5 to 14 ring-constituting atoms, which contains N as a ring-constituting atom and has an imide structure in which two carbonyl groups are bonded to the N. Specific examples include succinimide, glutarimide, and phthalimide.

[0089] As used herein, "optionally substituted by" and "substituted by" mean "optionally substituted by one substituent" and "substituted by one substituent," respectively, unless the number of substituents is specified (e.g., "one or more," "1 to 3," "1 or 2," "2," or "1"). For example, "B optionally substituted by A" and "B substituted by A" mean "B optionally substituted by one A" and "B substituted by one A," respectively.

[0090] In one aspect of the present invention, the compound of formula (I) or a salt thereof preferably comprises R 7 and R 8 is a hydrogen atom and X is 2,6-dichloro-4-pyridyl. Dichloropyridinylmethyl ester represented by the following formula may be hereinafter referred to as "DCPE" in this specification. In one aspect of the present invention, the compound represented by formula (I) or a salt thereof is 7 and R 8 is a hydrogen atom and X is trifluoromethyl. The 2,2,2-trifluoroethyl ester represented by the following formula may be referred to as "TEE" in this specification.

[0091] [ka]

[0092] [ka]

[0093] In one aspect of the present invention, a compound of formula (I) can be converted to a compound of formula (V):

[0094] [ka]

[0095] [In the formula, R 1 is a hydrogen atom, and R x is 2-methylpropyl, or octyl, or R 1 is 2-methylpropyl, and R x is methyl] Compounds represented by the following formula are excluded.

[0096] In addition, in this specification, the main chain amino group of the compound represented by formula (I) may be protected with a general protecting group such as an Fmoc group or a Boc group. In one aspect of the present invention, a compound of formula (IV):

[0097] [ka]

[0098] [In the formula, R 1 , R 2 , R 7 , R 8 and X is as defined herein, 7 and R 8 is a hydrogen atom, X is 2,6-dichloropyridin-4-yl, R 1 and R 2 is a hydrogen atom, or R 1is a hydrogen atom, and R 2 is methyl, ethyl, isopropyl, n-butyl, tert-butyl, or isobutyl, or R 1 is ethyl and R 2 is methyl, or ethyl). In this specification, salts of the compound represented by formula (I) include acid addition salts or base addition salts. Examples of acid addition salts include hydrochloride, hydrobromide, hydroiodide, phosphate, phosphonate, sulfate, etc.; sulfonates such as methanesulfonate, ethanesulfonate, benzenesulfonate, and p-toluenesulfonate; and carboxylates such as acetate, citrate, malate, tartrate, succinate, salicylate, maleate, fumarate, benzoate, malonate, glycolate, oxalate, glucuronate, adipate, glutarate, ketoglutarate, and hippurate. Examples of base addition salts include alkali metal salts such as sodium salt and potassium salt, alkaline earth metal salts such as magnesium salt and calcium salt, ammonium salts such as ammonium salt, alkylammonium salt, dialkylammonium salt, trialkylammonium salt, and tetraalkylammonium salt, and amino acid salts such as lysine salt, arginine salt, glycine salt, valine salt, threonine salt, serine salt, proline salt, and alanine salt, etc. These salts are produced by contacting the compound with an acid or base that can be used in the production of pharmaceuticals.

[0099] In the present specification, the compound represented by formula (I) or a salt thereof may be an anhydrate or may form a solvate such as a hydrate. As used herein, the term "solvate" refers to a solid in which the compound molecule and the solvent molecule form a complex, for example, when the solvent is water, it is called a hydrate. Solvates other than hydrates include solids containing alcohols (e.g., methanol, ethanol, n-propanol), dimethylformamide, etc.

[0100] The compounds of formula (I) and their salts can exist in several tautomeric forms, such as keto and enol forms, imine and enamine forms, and mixtures thereof. Tautomers exist as a mixture of tautomers in solution. In solid form, one tautomer usually predominates. Although one tautomer may be described, the present invention includes all tautomers of the compounds of the present invention.

[0101] The present invention includes all stereoisomers of the compounds of formula (I), such as enantiomers, diastereomers (including cis and trans geometric isomers), racemates, and other mixtures of such isomers. For example, the compounds of the present invention may have one or more asymmetric centers, and the compounds of the present invention include racemic mixtures, diastereomeric mixtures, and enantiomers of such compounds.

[0102] When the compound represented by formula (I) is obtained in a free form, it can be converted into a salt which the compound may form, or a hydrate or solvate thereof, in a conventional manner.

[0103] Furthermore, when the compound represented by formula (I) is obtained as a salt, hydrate, or solvate of the compound, it can be converted into the free form of the compound by a conventional method. The elements constituting the compound represented by formula (I) may be any isotope, and the present invention encompasses compounds of formula (I) containing isotopes. An isotope of the compound is one in which at least one atom has been replaced with an atom having the same atomic number (number of protons) but a different mass number (sum of the number of protons and neutrons). Examples of isotopes contained in the compound of the present invention include a hydrogen atom, a carbon atom, a nitrogen atom, an oxygen atom, a phosphorus atom, a sulfur atom, a fluorine atom, and a chlorine atom, each of which is 2 H, 3 H, 13 C, 14 C, 15 N, 17 O, 18 O, 31 P, 32 P, 35 S,18 F, 36 Cl, etc. In particular, 3 H and 14 Radioisotopes that decay by emitting radioactivity, such as C, are useful in testing the tissue distribution of pharmaceuticals or compounds in the body. Stable isotopes do not decay, their abundance remains almost constant, and they are not radioactive, so they can be used safely. The isotopes of the compounds of the present invention can be converted in accordance with conventional methods by replacing the reagents used in synthesis with reagents containing the corresponding isotope.

[0104] In one aspect of the present invention, the compound represented by formula (I) can be used as a reagent in an acylation reaction. Examples of the acylation reaction include acylation of a hydroxyl group or an amino group which may have a substituent. In one aspect of the present invention, the compound represented by formula (I) can be used as an activated ester for use in a peptide synthesis reaction. Peptide synthesis can be carried out by well-known techniques.

[0105] In another aspect of the present invention, the compound of formula (I) can be used as an activated ester for modifying groups, such as hydroxy or optionally substituted amino groups, contained in peptides or proteins by acylation.

[0106] In one aspect of the present invention, a compound represented by formula (I) can be used as a reagent for the aminoacylation of tRNA. The compound represented by formula (I) is an ester obtained by reacting an amino acid represented by formula (II) with a compound represented by formula (III). The amino acid represented by formula (II) includes not only natural amino acids but also unnatural amino acids. The present inventors have discovered that by reacting the above ester with tRNA, the amino acid represented by formula (II) can be attached to the 3' end of tRNA (aminoacylation reaction). This reaction can be carried out by known methods. In one embodiment, the aminoacylation of tRNA can be carried out in the presence of a catalyst or enzyme, preferably in the presence of Flexizyme. Flexizyme is an artificial aminoacylation RNA catalyst that recognizes and activates only the A (adenosine residue) of CCA at the 3' end of all tRNAs. That is, since Flexizyme does not have strict substrate specificity, it is possible to bind various amino acids, including unnatural amino acids, and amino acid derivatives to any tRNA (aminoacylation of tRNA), and to synthesize unique peptides incorporating unnatural amino acids. Examples of such Flexizymes include: Original Flexizyme Fx [5'-GGAUCGAAAGAUUUCCGCAGGCCCGAAAGGGUAUUGGCGUUAGGU-3', 45nt] (SEQ ID NO: 1), Enhanced Flexizyme eFx [5'-GGAUCGAAAGAUUUCCGCGGCCCCGAAAGGGGAUUAGCGUUAGGU-3', 45nt]) (SEQ ID NO: 2), Dinitrobenzyl Flexizyme dFx [5'-GGAUCGAAAGAUUUCCGCAUCCCCGAAAGGGUACAUGGCGUUAGGU-3', 46nt] (SEQ ID NO: 3), Aminoflexizyme aFx [5'-GGAUCGAAAGAUUUCCGCACCCCCGAAAGGGGUAAGUGGCGUUAGGU-3', 47 nt] (SEQ ID NO: 4) and the like are known (WO 2011 / 049157), but the present invention is not limited to these, and any substance having flexizyme activity can be suitably used. Although not limited thereto, dFx can be suitably used for DCPE, and eFx can be suitably used for TEE.

[0107] In one aspect of the present invention, a peptide synthesis method is provided, comprising synthesizing corresponding peptides from mRNA using an aminoacylated tRNA synthesized by the aminoacylation reaction described above in a cell-free translation system. Furthermore, in another aspect of the present invention, a peptide library preparation method is provided, comprising preparing an mRNA library and synthesizing peptides corresponding to each mRNA from the mRNA library using an cell-free translation system using the aminoacylated tRNA synthesized by the aminoacylation reaction described above, thereby preparing a peptide library. Here, the mRNA library may be prepared by purchasing it from a commercial source or by preparation by hand. For example, when preparing an mRNA library, a DNA library may be obtained according to the method described in Chemistry & Biology 18, 1562-1570 (2011) and / or Chemistry & Biology 21, 766-774 (2014), and the mRNA library may be prepared by transcribing the DNA library in a test tube.

[0108] As used herein, the term "cell-free translation system" is not particularly limited as long as it does not contain cells. In one aspect of the present invention, a system that synthesizes a target peptide or protein in a test tube by utilizing a protein synthesis function extracted from cells can be used as the cell-free translation system. Examples of cell-free translation systems that can be used include Escherichia coli extract, wheat germ extract, rabbit erythrocyte extract, and insect cell extract. In one aspect of the present invention, a reconstituted cell-free translation system can be used, constructed by reconstituting purified ribosomal proteins, aminoacyl-tRNA synthetases (aaRS), ribosomal RNA, amino acids, rRNA, GTP, ATP, translation initiation factors (IFs), elongation factors (EFs), release factors (RFs), and ribosome recycling factors (RRFs), as well as other factors required for translation.

[0109] As used herein, the cell-free translation system may also include an RNA polymerase for simultaneous transcription from DNA. In one aspect of the present invention, the cell-free translation system used may be a commercially available system, such as an Escherichia coli-derived system such as RTS-100 (registered trademark) from Roche Diagnostics, a reconstituted translation system such as PURESYSTEM (registered trademark) from PGI or PURExpress® In Vitro Protein Synthesis Kit from New England BioLabs, or a wheat germ extract-based system such as those from Zoigene or Cell Free Sciences.

[0110] Furthermore, as a system using Escherichia coli ribosomes, the techniques described in the following documents are known, for example: H.F. Kung et al., 1977, The Journal of Biological Chemistry, Vol. 252, No. 19, 6889-6894; M.C. Konza et al., 1985, Proceedings of the National Academy of Sciences of the United States of America, Vol. 82, 1648-1652; M.Y. Pavlov and M. Ehrenberg, 1996, Archives of Biochemistry and Biophysics, Vol. 328, No. 1, 9-16; Y. Shimizu et al., 2001, Nature Biotechnology, Vol. 19, No. 8, 751-755; H. Ohashi et al., 2007, Biochemical and Biophysical Research Communications, Vol. 352, No. 1, 270-276. The cell-free translation system allows expression products to be obtained in a highly pure form without purification. Expression of peptides using a cell-free translation system can be carried out using the Flexible Invitro Translation system (FIT system) in accordance with the method described in Goto, Y., Katoh, T. & Suga, H. Flexizymes for genetic code reprogramming. Nat Protoc 6, 779-790, (2011).

[0111] In one aspect of the present invention, a compound of formula (II):

[0112] [ka]

[0113] with a compound represented by formula (III):

[0114] [ka]

[0115] wherein L is a leaving group. In one embodiment of the present invention, the leaving group is a halogen atom, and R D- The group represented by SO2O- is selected from, for example, a chlorine atom, a bromine atom, an iodine atom, methanesulfonyloxy, benzenesulfonyloxy, toluenesulfonyloxy, trifluoromethanesulfonyloxy, pentafluoroethanesulfonyloxy, etc. The method can be carried out by a method known to those skilled in the art.

[0116] In one embodiment of the invention, the process is carried out in a solvent, such as DMF. In one embodiment of the invention, the process is carried out in the presence of a base, such as diisopropylethylamine.

[0117] In one embodiment of the present invention, the compound represented by formula (III) can be used in an amount of 0.45 to 1.45 equivalents, 0.65 to 1.25 equivalents, preferably 0.85 to 1.05 equivalents relative to the compound represented by formula (II).

[0118] In one embodiment of the present invention, the reaction temperature is set in the range of 0 to 40° C., 0 to 30° C., and preferably 0 to 25° C. In one embodiment of the present invention, the reaction time is set in the range of 30 minutes to 24 hours, 30 minutes to 12 hours, 30 minutes to 6 hours, and preferably 30 minutes to 2 hours.

[0119] In one aspect of the present invention, an activated amino acid ester is provided that is highly safe in terms of dangers when used as a reagent, such as ignition and explosion. The activated ester of the present invention is not a nitro compound or a cyanide compound, and is therefore considered to be safer than DBE or CME, which have been used as activated amino acid esters for the aminoacylation of tRNA. Evaluation tests were conducted to support this claim. Differential scanning calorimetry (DSC) can be used to determine the danger of an activated amino acid ester (Akiyoshi et al., NetsuSokutei 2018, 45(4), 161-167.; Sakira Kaneko, NetsuSokutei 1995, 22(1), 36-43.). DSC measures the reaction onset temperature (T DSC ) and the heat of reaction (Q DSC Because DSC can measure the heat release (exothermic decomposition energy) of a small sample, it is an effective tool for assessing the hazards of self-reactive substances. A correlation between the calorific value (exothermic decomposition energy) obtained by DSC and the fire and explosive properties of chemicals has been recognized, and DSC has traditionally been used as a hazard assessment test. In Japan, DSC is used to determine whether a chemical falls under the category of Class 5 hazardous materials (self-reactive substances) under the Fire Service Act. Internationally, the UN Recommendation on the Transport of Dangerous Goods (TDG) first uses the exothermic decomposition energy obtained by DSC to determine whether a chemical is an explosive or not. If, for example, a chemical is determined to be a Class 5 hazardous material, careful and complex storage and handling measures are required, such as avoiding heat, impact, and friction, separating it from flammable materials, avoiding contact with other chemicals, storing it in a cool, dark place, and ensuring that the container is not damaged, resulting in increased storage and transportation costs.

[0120] As a test method for determining the 5th class of hazardous materials (self-reactive substances) under the Fire Service Act, 2,4-dinitrotoluene and benzoyl peroxide are used as standard compounds. DSC and T DSC One method is to plot the reference points of each of the above and use a straight line connecting the two points as the risk assessment line. If the substance is on or above the assessment line, it is deemed "risky" (class 5 hazardous material), and if it is below the assessment line, it is deemed "not risky" (not class 5 hazardous material). The present inventors performed the above method on DBE, which has previously been used as an activated ester of an amino acid for aminoacylation of tRNA, and DCPE and TEE of the present invention. As a result, DBE was determined to be class 5 hazardous material, while DCPE and TEE were determined not to be class 5 hazardous material (Test Example). This demonstrates that the activated ester of the present invention has reduced safety and waste risks, is easy to handle, and can reduce disposal costs compared to conventional activated esters. Example

[0121] Example 1: Synthesis of activated esters of amino acids The present invention will be explained in more detail below by showing synthesis examples as examples, but the present invention is not limited to these examples.

[0122] In the present specification, when amino acids and the like are represented by abbreviations, each representation is based on the abbreviations according to the IUPAC-IUB Commission on Biochemical Nomenclature or on the abbreviations commonly used in the art.

[0123] The abbreviations used in the synthesis examples are as follows: Fmoc as 9-fluorenylmethyloxycarbonyl or 9-fluorenylmethoxycarbonyl; Boc as tert-butoxycarbonyl; tBu as tertiary butyl; DMF as N,N-dimethylformamide; DIPEA as diisopropylethylamine; EtOAc as ethyl acetate; Sodium sulfate as Na2SO4; DCM as dichloromethane; MTHP as 4-methyltetrahydropyran; MTBE as methyl tertiary butyl ether; CPME as cyclopentyl methyl ether; Flexizyme Fx: THF as tetrahydrofuran; HCl as hydrochloric acid; milliliters (units) as mL; M as molar (unit: mol / L); mM as millimolar (unit); mm as millimeters (unit); nm as nanometers (units); μm as micrometer (unit); Angstroms (units) as Å; Minutes (units) as min; MS as mass spectrometry; mmol as millimoles (units); mg as milligrams (units); Liquid chromatography-mass spectrometry (LC-MS or LC / MS); and tR as retention time.

[0124] Unless otherwise specified, proton nuclear magnetic resonance (H NMR) spectra of the following synthesis examples were measured in a deuterated chloroform or deuterated dimethyl sulfoxide solvent using a JEOL JNM-ECP300 or a JEOL JNM-ECX300, or a Bruker Ascend™500, and chemical shifts are shown as δ values ​​(ppm) using tetramethylsilane as the internal standard (0.0 ppm).

[0125] In describing NMR spectra, "s" means singlet, "d" means doublet, "t" means triplet, "q" means quartet, "dd" means doublet of doublets, "dt" means doublet of triplets, "sept" means septet, "m" means multiplet, "br" means broad, "J" means coupling constant, "Hz" means Hertz, "CDCl3" means deuterated chloroform, and "DMSO-d6" means deuterated dimethyl sulfoxide.

[0126] Unless otherwise specified, high-performance liquid chromatography / mass spectrometry was measured using either a Waters ACQUITY UPLC H-Class / QDa, a Waters ACQUITY UPLC H-Class / SQD2, or a Shimadzu LC-20AD / Triple Tof5600.

[0127] In the description of high performance liquid chromatography / mass spectrometry, ESI+ is the positive mode of electrospray ionization, M+H means proton adduct, and M+Na means sodium adduct.

[0128] In the description of high performance liquid chromatography / mass spectrometry, ESI- stands for negative mode electrospray ionization, and MH stands for proton-deficient. The purity of the activated esters of amino acids synthesized according to the following synthesis examples was calculated from the area ratio of the LC / MS chromatograms under the following analytical conditions A to D, and mass spectrometry was performed using a single quadrupole mass spectrometer and an ESI-MS(+) ion source.

[0129] Analysis conditions A Column: Kinetex® EVO C18, 2.6 μm, 2.1 x 150 mm, 100 Å Mobile phase A: 0.025% TFA in H2O Mobile phase B:0.025% TFA in CH3CN Column temperature: 60℃ Gradient (%B): 0-40% over 7.15 min, then 40% from 7.16 to 9.00 min, then 95-95% over 1.55 min, flow rate: 0.5 mL / min Detection: UV 220nm. Analysis conditions B Column: Kinetex® EVO C18, 2.6 μm, 2.1 x 150 mm, 100 Å Mobile phase A: 0.025% TFA in H2O Mobile phase B:0.025% TFA in CH3CN Column temperature: 60℃ Gradient (%B): 5-95% over 7.15 min, then 95% from 7.16 min to 10.55 min, flow rate: 0.5 mL / min Detection: UV 220nm. Analysis conditions C Column: Waters XBridge BEH C18 2.5 μm 2.1 × 150 mm 130 Å Mobile phase A: 0.025% TFA in H2O Mobile phase B:0.025% TFA in CH3CN Column temperature: 60℃ Gradient (%B): 0-40% over 7.15 min, then 40% from 7.16 to 9.00 min, then 95-95% over 1.55 min, flow rate: 0.5 mL / min Detection: UV 220nm. Analysis conditions D Column: Waters ACQUITY UPLC BEH C18 1.7μm 2.1 x 50mm, 130Å Mobile phase A: 0.025% TFA in H2O Mobile phase B:0.025% TFA in CH3CN Column temperature: 60℃ Gradient (%B): 5 - 95% over 2.78 min, then 95% from 2.78 min to 3.61 min, flow rate: 0.4 mL / min Detection: UV 220nm. The structures of activated esters of amino acids synthesized according to the following synthesis examples and the results of LC / MS analysis are shown in Table 1.

[0130] Example 1-1: 2,2,2-trifluoroethyl L-phenylalaninate hydrochloride (Compound No. 39)

[0131] [ka]

[0132] To a flask equipped with a nitrogen balloon, (tert-butoxycarbonyl)-L-phenylalanine (7.96 g, 30.0 mmol), DMF (60.0 mL), 2,2,2-trifluoroethyl trifluoromethanesulfonate (6.61 g, 28.5 mmol), and DIPEA (6.29 mL, 36.0 mmol) were added in this order. After stirring at room temperature, the reaction was quenched by the addition of 1 M aqueous HCl. The mixture was extracted with EtOAc, and the organic layer was washed with saturated brine, dried over anhydrous Na2SO4, filtered through a glass filter, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; EtOAc / hexane = 0 / 100 → 100 / 0). To the resulting compound, DCM (30.0 mL) and 4 N HCl / MTHP (30.0 mL) were added in this order. After stirring at room temperature, the mixture was concentrated under reduced pressure. The residue was added with MTBE, filtered, washed with MTBE, and dried in vacuo to give 2,2,2-trifluoroethyl L-phenylalaninate hydrochloride (7.28 g, 25.7 mmol, 86% yield) as a colorless solid.

[0133] Example 1-2: 2,2,2-trifluoroethyl(2-chloroacetyl)-L-phenylalaninate (Compound No. 90)

[0134] [ka]

[0135] 2,2,2-Trifluoroethyl L-phenylalaninate hydrochloride (2.84 g, 10.0 mmol), THF (25.0 mL), N-methylmorpholine (2.42 mL, 22.0 mmol), and chloroacetyl chloride (0.881 mL, 11.0 mmol) were added sequentially to a flask equipped with a nitrogen balloon while maintaining the temperature below −15°C. After stirring at −15°C, the reaction was quenched by the addition of 1 M HCl. The mixture was extracted with EtOAc, and the organic layer was washed with saturated brine, dried over anhydrous Na2SO4, filtered through a glass filter, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; EtOAc / hexane = 0 / 100 → 100 / 0) to give 2,2,2-trifluoroethyl(2-chloroacetyl)-L-phenylalaninate (3.00 g, 9.27 mmol, 93% yield) as a colorless solid.

[0136] Example 1-3: (2,6-Dichloropyridin-4-yl)methylacetyl-L-alaninate (Compound No. 87)

[0137] [ka]

[0138] Acetyl-L-alanine (2.62 g, 20.0 mmol), DMF (40.0 mL), 4-(bromomethyl)-2,6-dichloropyridine (4.58 g, 19.0 mmol), and DIPEA (4.19 mL, 24.0 mmol) were added sequentially to a flask equipped with a nitrogen balloon. After stirring at room temperature, the reaction was quenched by the addition of 2 M aqueous HCl. The mixture was extracted with EtOAc, and the organic layer was washed with saturated brine, dried over anhydrous Na2SO4, filtered through a glass filter, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; EtOAc / hexane = 0 / 100 → 100 / 0) to give (2,6-dichloropyridin-4-yl)methylacetyl-L-alaninate (2.93 g, 10.1 mmol, 50% yield) as a colorless solid.

[0139] Example 1-4: (2,6-dichloropyridin-4-yl)methyl(2-([1,1'-biphenyl]-4-yl)ethyl)glycinate hydrochloride (Compound No. 35)

[0140] [ka]

[0141] A flask equipped with a nitrogen balloon was charged with (2-([1,1'-biphenyl]-4-yl)ethyl)glycine (1.55 g, 6.07 mmol, CAS Registry Number: 1906593-76-7), 1,4-dioxane (14.0 mL), HO (7.00 mL), sodium bicarbonate (1.53 g, 18.2 mmol), and di-tert-butyl dicarbonate (1.32 g, 6.07 mmol, CAS Registry Number: 24424-99-5) in this order under ice cooling. After stirring at room temperature, the reaction was quenched by the addition of aqueous citric acid. The mixture was extracted with EtOAc, and the organic layer was washed with saturated brine, dried over anhydrous NaSO, filtered through a glass filter, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; 0.1% aqueous formic acid / acetonitrile = 50 / 50 → 20 / 80). To the resulting compound (1.00 g), DMF (5.6 mL), 4-(bromomethyl)-2,6-dichloropyridine (0.644 g, 2.67 mmol), and DIPEA (0.590 mL, 3.38 mmol) were added in this order. After stirring at room temperature, the reaction was quenched by adding 1 M aqueous HCl. The mixture was extracted with EtOAc, and the organic layer was washed successively with aqueous sodium bicarbonate and saturated brine, dried over anhydrous Na2SO4, filtered through a glass filter, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; EtOAc / hexane = 20 / 80 → 40 / 60). To the resulting compound, DCM (8.0 mL) and 4 N HCl / MTHP (8.0 mL) were added in this order. After stirring at room temperature, the mixture was concentrated under reduced pressure. Diisopropyl ether was added to the residue, filtered, washed with diisopropyl ether, and dried in vacuo to give (2,6-dichloropyridin-4-yl)methyl(2-([1,1'-biphenyl]-4-yl)ethyl)glycinate hydrochloride (941 mg, 2.08 mmol) as a colorless solid.

[0142] Example 1-5: (2,6-Dichloropyridin-4-yl)methyl (S)-2-amino-3-(3-guanidinophenyl)propanoate dihydrochloride (Compound No. 74)

[0143] [ka]

[0144] To a flask equipped with a nitrogen balloon, (S)-3-(3-aminophenyl)-2-((tert-butoxycarbonyl)amino)propanoic acid (1.03 g, 3.67 mmol), absolute ethanol (15 mL), and N,N'-bis(tert-butoxycarbonyl)-1H-pyrazole-1-carboxamidine (1.14 g, 3.67 mmol, CAS Registry Number: 152120-54-2) were added in that order. After stirring at room temperature, the reaction was quenched by the addition of 1 M aqueous HCl. The mixture was extracted with EtOAc, and the organic layer was washed with saturated brine, dried over anhydrous Na2SO4, filtered through a glass filter, and concentrated under reduced pressure. To the resulting residue, DMF (60.0 mL), 4-(bromomethyl)-2,6-dichloropyridine (0.841 g, 3.49 mmol), and DIPEA (0.770 mL, 4.41 mmol) were added in that order. After stirring at room temperature, the reaction was quenched by adding 1 M aqueous HCl. The mixture was extracted with EtOAc, and the organic layer was washed successively with aqueous sodium bicarbonate and saturated brine, dried over anhydrous Na2SO4, filtered through a glass filter, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; EtOAc / hexane = 10 / 90 → 30 / 70). To the resulting compound, DCM (26.0 mL) and 4 N HCl / MTHP (26.0 mL) were added in that order. After stirring at room temperature, the mixture was concentrated under reduced pressure. Diethyl ether was added to the residue, filtered, washed with diethyl ether, and dried in vacuo to give (2,6-dichloropyridin-4-yl)methyl (S)-2-amino-3-(3-guanidinophenyl)propanoate dihydrochloride (962 mg, 2.11 mmol) as a colorless solid.

[0145] Example 1-6: (2,6-Dichloropyridin-4-yl)methyl (S)-3-cyclopentyl-2-(methylamino)propanoate hydrochloride (Compound No. 75)

[0146]

change

[0147] To a flask equipped with a nitrogen balloon, (S)-2-((tert-butoxycarbonyl)amino)-3-cyclopentylpropanoic acid (10.0 g, 38.9 mmol, CAS Registry Number: 143415-31-0), THF (300 mL), and 60% sodium hydride (2.80 g, 117 mmol, CAS Registry Number: 7646-69-7) were added sequentially under ice-cooling. The mixture was stirred at room temperature for 30 minutes, and then iodomethane (11.0 g, 77.7 mmol, CAS Registry Number: 74-88-4) was added. The mixture was stirred at room temperature overnight, and then aqueous citric acid was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; acetonitrile / 0.1% formic acid aqueous solution = 10 / 90 → 80 / 20) to give (S)-2-((tert-butoxycarbonyl)(methyl)amino)-3-cyclopentylpropanoic acid (5.00 g). To the resulting compound, DMF (25 mL), 4-(bromomethyl)-2,6-dichloropyridine (4.30 g, 17.9 mmol, CAS Registry Number: 175204-45-2), and DIPEA (3.92 mL, 22.6 mmol) were added in that order. The mixture was stirred at room temperature for 3 hours, and then ethyl acetate was added to the mixture. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; acetonitrile / 0.1% formic acid aqueous solution = 10 / 90 → 100 / 0) to give (2,6-dichloropyridin-4-yl)methyl (S)-2-((tert-butoxycarbonyl)(methyl)amino)-3-cyclopentylpropanoate (6.67 g). To the obtained compound (3.00 g), dichloromethane (26 mL) and 4 M hydrochloric acid / MTHP (26 mL) were added in sequence under ice-cooling. The mixture was stirred at room temperature for 2 hours, and then concentrated under reduced pressure.The residue was suspended and washed with diethyl ether and dried in vacuo to give (2,6-dichloropyridin-4-yl)methyl (S)-3-cyclopentyl-2-(methylamino)propanoate hydrochloride (2.62 g, 7.13 mmol) as a colorless solid.

[0148] Example 1-7: 2,2,2-trifluoroethyl (S)-2-amino-3-(6-phenylpyridin-3-yl)propanoate dihydrochloride (Compound No. 76)

[0149] [ka]

[0150] To a flask equipped with a nitrogen balloon, zinc (9.22 g, 141 mmol, CAS Registry Number: 7440-66-6), DMF (150 mL), and iodine (1.79 g, 7.05 mmol, CAS Registry Number: 7553-56-2) were added in that order. After stirring the mixture at room temperature for 10 minutes, (S)-methyl 2-(tert-butoxycarbonylamino)-3-iodopropanoate (15.5 g, 47.0 mmol, CAS Registry Number: 93267-04-0) and iodine (1.79 g, 7.05 mmol, CAS Registry Number: 7553-56-2) were added in that order. After stirring the mixture at room temperature for 30 minutes, 5-bromo-2-phenylpyridine (13.2 g, 56.4 mmol, CAS Registry Number: 27012-25-5), 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (0.96 g, 2.3 mmol, CAS Registry Number: 657408-07-6), and tris(dibenzylideneacetone)dipalladium-chloroform (1.46 g, 1.41 mmol, CAS Registry Number: 52522-40-4) were sequentially added to the mixture. After stirring at 50°C for 3 hours, the mixture was filtered, extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 10 → 1 / 3) to give methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(6-phenylpyridin-3-yl)propanoate (16.0 g). To the resulting compound, isopropanol (180 mL), water (60 mL), calcium chloride (79.7 g, 718 mmol, CAS Registry Number: 10043-52-4), and lithium hydroxide monohydrate (7.55 g, 180 mmol, CAS Registry Number: 1310-66-3) were added sequentially under ice-cooling. The mixture was stirred at room temperature for 24 hours, and then sodium dihydrogen phosphate was added to quench the reaction. The mixture was filtered, extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure.The residue was purified by flash column chromatography (silica gel; methanol / dichloromethane = 1 / 50 → 1 / 10) to give (S)-2-((tert-butoxycarbonyl)amino)-3-(6-phenylpyridin-3-yl)propanoic acid (14.1 g). To the resulting compound, DMF (140 mL), 2,2,2-trifluoroethyl trifluoromethanesulfonate (9.08 g, 39.1 mmol, CAS Registry Number: 6226-25-1), and DIPEA (6.39 g, 49.4 mmol) were added sequentially. The mixture was stirred at room temperature for 3 hours, and then water and sodium dihydrogen phosphate were added sequentially to quench the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 10 → 1 / 1) to give 2,2,2-trifluoroethyl (S)-2-((tert-butoxycarbonyl)amino)-3-(6-phenylpyridin-3-yl)propanoate (15.1 g). To the obtained compound (1.87 g), dichloromethane (30 mL) and 4M hydrochloric acid / MTHP (30 mL) were added, successively. The mixture was stirred at room temperature for 3 hours, and then concentrated under reduced pressure. The residue was suspended and washed with diethyl ether and dried in vacuo to give 2,2,2-trifluoroethyl (S)-2-amino-3-(6-phenylpyridin-3-yl)propanoate dihydrochloride (1.56 g, 4.32 mmol) as a colorless solid.

[0151] Example 1-8: (2,6-Dichloropyridin-4-yl)methyl (S)-2-amino-4-(quinolin-5-yl)butanoate dihydrochloride (Compound No. 77)

[0152] [ka]

[0153] To a flask equipped with a nitrogen balloon, zinc (47.2 g, 721 mmol, CAS Registry Number: 7440-66-6), DMF (500 mL), and iodine (36.6 g, 144 mmol, CAS Registry Number: 7553-56-2) were added in that order. After stirring the mixture at room temperature for 5 minutes, methyl (S)-2-((tert-butoxycarbonyl)amino)-4-iodobutanoate (99.0 g, 288 mmol, CAS Registry Number: 101650-14-0) was added. After stirring the mixture at room temperature for 1 hour, 5-bromoquinoline (50.0 g, 240 mmol, CAS Registry Number: 4964-7-10), 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (9.85 g, 24.0 mmol, CAS Registry Number: 657408-07-6), and tris(dibenzylideneacetone)dipalladium-chloroform (11.0 g, 12.0 mmol, CAS Registry Number: 52522-40-4) were sequentially added to the mixture. After stirring at 50°C for 3 hours, the mixture was filtered and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 1) to give methyl (S)-2-((tert-butoxycarbonyl)amino)-4-(quinolin-5-yl)butanoate (40.0 g). To the resulting compound, THF (200 mL) and lithium hydroxide monohydrate (5.85 g, 139 mmol, CAS Registry Number: 1310-66-3) dissolved in water (200 mL) were added sequentially under ice-cooling. The mixture was stirred at room temperature for 2 hours, and then aqueous sodium dihydrogen phosphate was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give (S)-2-((tert-butoxycarbonyl)amino)-4-(quinolin-5-yl)butanoic acid (20.0 g).To the resulting compound (5.00 g), DMF (50 mL), 4-(bromomethyl)-2,6-dichloropyridine (3.46 g, 14.4 mmol, CAS reg. no.: 175204-45-2), and DIPEA (3.17 mL, 18.2 mmol) were added, in that order. The mixture was stirred at room temperature for 30 minutes, and then aqueous sodium dihydrogen phosphate was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 5) to give (2,6-dichloropyridin-4-yl)methyl (S)-2-((tert-butoxycarbonyl)amino)-4-(quinolin-5-yl)butanoate (6.31 g). To the resulting compound (3.30 g), dichloromethane (25 mL) and 4M hydrochloric acid / MTHP (25 mL) were added in sequence. The mixture was stirred at room temperature for 2 hours, and then concentrated under reduced pressure. The residue was suspended in diethyl ether and dried in vacuo to give (2,6-dichloropyridin-4-yl)methyl (S)-2-amino-4-(quinolin-5-yl)butanoate dihydrochloride (2.92 g, 6.84 mmol) as a colorless solid.

[0154] Example 1-9: (2,6-Dichloropyridin-4-yl)methyl (S)-2-amino-3-(6-aminopyridin-3-yl)propanoate dihydrochloride (Compound No. 78)

[0155] [ka]

[0156] To a flask equipped with a nitrogen balloon, zinc (22.7 g, 347 mmol, CAS Registry Number: 7440-66-6), DMF (400 mL), and iodine (8.80 g, 34.7 mmol, CAS Registry Number: 7553-56-2) were added in this order. After stirring the mixture at room temperature, (S)-methyl 2-(tert-butoxycarbonylamino)-3-iodopropanoate (41.9 g, 127 mmol, CAS Registry Number: 93267-04-0) and iodine (8.80 g, 34.7 mmol, CAS Registry Number: 7553-56-2) were added in this order. After stirring the mixture at room temperature for 30 minutes, 2-amino-5-bromopyridine (20.0 g, 116 mmol, CAS Registry Number: 1072-97-5), 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (9.49 g, 23.1 mmol, CAS Registry Number: 657408-07-6), and tris(dibenzylideneacetone)dipalladium-chloroform (5.29 g, 5.78 mmol, CAS Registry Number: 52522-40-4) were sequentially added to the mixture. After stirring at 50°C for 3 hours, ethyl acetate was added to the mixture, which was then filtered and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 1) to give methyl (S)-3-(6-aminopyridin-3-yl)-2-((tert-butoxycarbonyl)amino)propanoate (30.0 g). To the resulting compound (40.0 g), dichloromethane (200 mL), tert-butanol (800 mL), di-tert-butyl dicarbonate (35.5 g, 163 mmol, CAS Registry Number: 24424-99-5), and sodium iodide (24.4 g, 163 mmol, CAS Registry Number: 7681-82-5) were added, in that order. The mixture was stirred at room temperature overnight, then concentrated under reduced pressure, ethyl acetate was added, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure.The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 1) to give methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(6-((tert-butoxycarbonyl)amino)pyridin-3-yl)propanoate (50.0 g). To the resulting compound (30.0 g), THF (100 mL), isopropanol (600 mL), water (200 mL), calcium chloride (135 g, 1210 mmol, CAS Registry Number: 10043-52-4), and lithium hydroxide monohydrate (12.8 g, 304 mmol, CAS Registry Number: 1310-66-3) were added in this order under ice cooling. The mixture was stirred at room temperature, and then sodium dihydrogen phosphate was added to quench the reaction. The mixture was filtered and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / dichloromethane = 1 / 1) to give (S)-2-((tert-butoxycarbonyl)amino)-3-(6-phenylpyridin-3-yl)propanoic acid (15.0 g). To the resulting compound (9.00 g), DMF (50 mL), 4-(bromomethyl)-2,6-dichloropyridine (5.40 g, 22.4 mmol, CAS Registry Number: 175204-45-2), and DIPEA (4.93 mL, 28.3 mmol) were added in that order. The mixture was stirred at room temperature for 2 hours, and then ethyl acetate was added to the mixture. The organic layer was washed successively with aqueous sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 5) to give (2,6-dichloropyridin-4-yl)methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(6-((tert-butoxycarbonyl)amino)pyridin-3-yl)propanoate (11.2 g). To the obtained compound (3.00 g), dichloromethane (25 mL) and 4 M hydrochloric acid / MTHP (25 mL) were added in that order.After stirring the mixture at room temperature, the mixture was concentrated under reduced pressure, the residue was suspended in methyl tert-butyl ether, washed, and dried in vacuo to give (2,6-dichloropyridin-4-yl)methyl (S)-2-amino-3-(6-aminopyridin-3-yl)propanoate dihydrochloride (2.32 g, 5.60 mmol) as a colorless solid.

[0157] Example 1-10: (2,6-Dichloropyridin-4-yl)methyl (S)-2-amino-4-(2-aminopyridin-4-yl)butanoate dihydrochloride (Compound No. 79)

[0158] [ka]

[0159] To a flask equipped with a nitrogen balloon, zinc (8.57 g, 131 mmol, CAS Registry Number: 7440-66-6), DMF (200 mL), and iodine (6.66 g, 26.2 mmol, CAS Registry Number: 7553-56-2) were added in that order. After stirring the mixture at room temperature for 5 minutes, methyl (S)-2-((tert-butoxycarbonyl)amino)-4-iodobutanoate (15.0 g, 43.7 mmol, CAS Registry Number: 101650-14-0) was added. After stirring the mixture at room temperature for 1 hour, 2-amino-4-bromopyridine (8.32 g, 48.1 mmol, CAS Registry Number: 84249-14-9), 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (3.59 g, 8.75 mmol, CAS Registry Number: 657408-07-6), and tris(dibenzylideneacetone)dipalladium-chloroform (2.00 g, 2.18 mmol, CAS Registry Number: 52522-40-4) were added sequentially to the mixture. After stirring at 60°C for 3 hours, the mixture was filtered, extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 2) to give methyl (S)-4-(2-aminopyridin-4-yl)-2-((tert-butoxycarbonyl)amino)butanoate (9.00 g). To the resulting compound, tert-butanol (90 mL), sodium iodide (5.23 g, 34.9 mmol, CAS Registry Number: 7681-82-5), and di-tert-butyl dicarbonate (7.62 g, 34.9 mmol, CAS Registry Number: 24424-99-5) were added, in that order. The mixture was stirred at room temperature for 16 hours, after which ethyl acetate was added to the mixture. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure.The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 2) to give methyl (S)-2-((tert-butoxycarbonyl)amino)-4-(2-((tert-butoxycarbonyl)amino)pyridin-4-yl)butanoate (8.80 g). To the resulting compound, isopropanol (120 mL), calcium chloride (38.2 g, 344 mmol, CAS Registry Number: 10043-52-4), and lithium hydroxide monohydrate (3.61 g, 86.0 mmol, CAS Registry Number: 1310-66-3) dissolved in water (40 mL) were added sequentially under ice cooling. The mixture was stirred at room temperature for 16 hours, then filtered, and the reaction was quenched by the addition of aqueous sodium dihydrogen phosphate solution. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 1) to give (S)-2-((tert-butoxycarbonyl)amino)-4-(2-((tert-butoxycarbonyl)amino)pyridin-4-yl)butanoic acid (8.00 g). To the resulting compound, DMF (80 mL), 4-(bromomethyl)-2,6-dichloropyridine (4.63 g, 19.2 mmol, CAS registration number: 175204-45-2), and DIPEA (4.23 mL, 24.3 mmol) were added, in that order. The mixture was stirred at room temperature for 30 minutes, and then aqueous sodium dihydrogen phosphate was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; acetonitrile / 10 mM aqueous ammonium bicarbonate solution = 10 / 90 → 100 / 0) to give (2,6-dichloropyridin-4-yl)methyl (S)-2-((tert-butoxycarbonyl)amino)-4-(2-((tert-butoxycarbonyl)amino)pyridin-4-yl)butanoate (5.62 g).To the resulting compound (3.00 g), dichloromethane (18 mL), triisopropylsilane (2.14 g, 13.5 mmol, CAS registration number: 6485-79-6), and 2,2,2-trifluoroacetic acid (12.5 mL) were added in this order. After stirring the mixture at room temperature for 18 hours, the mixture was concentrated under reduced pressure, and 4M hydrochloric acid / CPME (25 mL) was added. After stirring the mixture at room temperature for 1 hour, the mixture was filtered, washed with diethyl ether, and dried in vacuo to obtain (2,6-dichloropyridin-4-yl)methyl (S)-2-amino-4-(2-aminopyridin-4-yl)butanoate dihydrochloride (1.52 g, 3.55 mmol) as a colorless solid.

[0160] Example 1-11: (S)-2-(4-(4-amino-5-((2,6-dichloropyridin-4-yl)methoxy)-5-oxopentanoyl)piperazin-1-yl)acetic acid dihydrochloride (Compound No. 60)

[0161] [ka]

[0162] To a flask equipped with a nitrogen balloon, (S)-5-(benzyloxy)-4-((tert-butoxycarbonyl)amino)-5-oxopentanoic acid (15.0 g, 44.5 mmol, CAS registration number: 30924-93-7), tert-butyl 2-(piperazin-1-yl)acetate (8.90 g, 44.5 mmol, CAS registration number: 112257-22-4), and THF (300 mL) were added in this order under ice cooling. After stirring the mixture at room temperature, 2-cyano-2-(hydroxyimino)ethyl acetate (6.94 g, 48.9 mmol, CAS Registry Number: 3849-21-6) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (9.38 g, 48.9 mmol, CAS Registry Number: 25952-53-8) were sequentially added to the mixture under ice-cooling. After stirring the mixture at room temperature overnight, ethyl acetate was added to the mixture. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 1) to give benzyl (S)-5-(4-(2-(tert-butoxy)-2-oxoethyl)piperazin-1-yl)-2-((tert-butoxycarbonyl)amino)-5-oxopentanoate (13.0 g). Methanol (260 mL) and palladium on carbon (2.60 g, CAS registration number: 7440-05-3) were added sequentially to the obtained compound. After stirring under a hydrogen atmosphere at room temperature for 2 hours, the mixture was filtered, extracted with ethyl acetate, and concentrated under reduced pressure to give (S)-5-(4-(2-(tert-butoxy)-2-oxoethyl)piperazin-1-yl)-2-((tert-butoxycarbonyl)amino)-5-oxopentanoic acid (12.0 g). To the obtained compound were added DMF (100 mL), 4-(bromomethyl)-2,6-dichloropyridine (6.39 g, 26.5 mmol, CAS registration number: 175204-45-2) and DIPEA (5.60 mL, 32.1 mmol) in that order.After stirring the mixture at room temperature for 2 hours, ethyl acetate was added to the mixture, and the organic layer was washed successively with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 1) to give (2,6-dichloropyridin-4-yl)methyl (S)-5-(4-(2-(tert-butoxy)-2-oxoethyl)piperazin-1-yl)-2-((tert-butoxycarbonyl)amino)-5-oxopentanoate (9.07 g). To the resulting compound (3.00 g), dichloromethane (25 mL) and 4M hydrochloric acid / CPME (25 mL) were added, successively. The mixture was stirred at room temperature for 3 hours, and then concentrated under reduced pressure. The residue was suspended and washed with methyl tert-butyl ether and dried in vacuo to give (S)-2-(4-(4-amino-5-((2,6-dichloropyridin-4-yl)methoxy)-5-oxopentanoyl)piperazin-1-yl)acetic acid dihydrochloride (2.39 g, 4.72 mmol) as a colorless solid.

[0163] Example 1-12: (2,6-Dichloropyridin-4-yl)methyl (3-methoxypropyl)glycinate hydrochloride (Compound No. 61)

[0164] [ka]

[0165] To a flask equipped with a nitrogen balloon, 3-methoxypropylamine (10.0 g, 112 mmol, CAS Registry Number: 5332-73-0), ethyl glyoxylate (23.0 mL, 225 mmol, CAS Registry Number: 924-44-7), and dichloromethane (200 mL) were added sequentially. After stirring the mixture at room temperature for 3 minutes, sodium cyanoborohydride (14.0 g, 223 mmol, CAS Registry Number: 25895-60-7) and acetic acid (9.5 mL) were added sequentially. After stirring the mixture at room temperature for 30 minutes, triethylamine (31.3 mL, 224 mmol) and di-tert-butyl dicarbonate (37.0 g, 170 mmol, CAS Registry Number: 24424-99-5) were added sequentially. After stirring the mixture at room temperature, the mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 1) to give ethyl N-(tert-butoxycarbonyl)-N-(3-methoxypropyl)glycinate (11.0 g). To the resulting compound (9.50 g), 1,4-dioxane (10 mL), water (10 mL), and lithium hydroxide monohydrate (2.30 g, 54.8 mmol) were added sequentially at room temperature. The mixture was stirred at room temperature, and then aqueous sodium dihydrogen phosphate was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give N-(tert-butoxycarbonyl)-N-(3-methoxypropyl)glycine (8.30 g). To the obtained compound (7.30 g), DMF (50 mL), 4-(bromomethyl)-2,6-dichloropyridine (5.70 g, 1.91 mmol, CAS registration number: 175204-45-2) and DIPEA (6.0 mL, 2.30 mmol) were added in this order.After stirring the mixture at room temperature, it was purified by flash column chromatography (silica gel; water / methanol = 95 / 5 → 0 / 100) to give (2,6-dichloropyridin-4-yl)methyl N-(tert-butoxycarbonyl)-N-(3-methoxypropyl)glycinate (6.18 g). To the obtained compound (1.90 g), dichloromethane (18 mL) and 4M hydrochloric acid / CPME (18 mL) were added, successively. After stirring the mixture at room temperature for 2 hours, the mixture was concentrated under reduced pressure. The residue was suspended and washed with diethyl ether and dried in vacuo to give (2,6-dichloropyridin-4-yl)methyl (3-methoxypropyl)glycinate (1.31 g, 3.81 mmol) as a colorless solid.

[0166] Example 1-13: 2,2,2-trifluoroethyl (S)-2-amino-3-(4-(5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)phenyl)propanoate hydrochloride (Compound No. 62)

[0167] [ka]

[0168] To a flask equipped with a nitrogen balloon, (S)-3-(4-aminophenyl)-2-((tert-butoxycarbonyl)amino)propanoic acid (7.01 g, 25 mmol, CAS Registry Number: 55533-24-9), DMF (50 mL), allyl (2,5-dioxopyrrolidin-1-yl)carbonate (4.88 g, 24.5 mmol, CAS Registry Number: 135544-68-2), and DIPEA (4.80 mL, 27.5 mmol) were added in this order under ice-cooling. The mixture was stirred at room temperature, and then hydrochloric acid was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed successively with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give (S)-3-(4-((allyloxy)carbonyl)amino)phenyl)-2-((tert-butoxycarbonyl)amino)propanoic acid. To the resulting compound were added DMF (50 mL), 2,2,2-trifluoroethyl trifluoromethanesulfonate (5.80 g, 25.0 mmol, CAS Registry Number: 6226-25-1), and DIPEA (5.24 mL, 30 mmol), successively. The mixture was stirred at room temperature, and then hydrochloric acid was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed successively with water and aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / heptane = 0 / 100 → 100 / 0) to give 2,2,2-trifluoroethyl (S)-3-(4-((allyloxy)carbonyl)amino)phenyl)-2-((tert-butoxycarbonyl)amino)propanoate (3.70 g). To the resulting compound (3.57 g), dichloromethane (20 mL), tetrakis(triphenylphosphine)palladium (0.185 g, 0.160 mmol, CAS Registry Number: 14221-01-3), and phenylsilane (2.95 mL, 24.0 mmol, CAS Registry Number: 694-53-1) were added in this order under a nitrogen atmosphere and ice cooling.After stirring under ice-cooling under a nitrogen atmosphere, the mixture was purified by flash column chromatography (silica gel; ethyl acetate / heptane=0 / 100→100 / 0) to give 2,2,2-trifluoroethyl (S)-3-(4-aminophenyl)-2-((tert-butoxycarbonyl)amino)propanoate (2.26 g). To the resulting compound (0.725 g), DMF (8.0 mL), biotin (0.537 g, 2.20 mmol, CAS Registry Number: 58-85-5), ethyl 2-cyano-2-(hydroxyimino)acetate (0.313 g, 2.20 mmol, CAS Registry Number: 3849-21-6), and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (0.422 g, 2.20 mmol, CAS Registry Number: 25952-53-8) were added in that order. The mixture was stirred at room temperature, and then water was added to quench the reaction. The resulting residue was collected by filtration, washed with water, and purified by flash column chromatography (silica gel; methanol / dichloromethane = 3 / 97 → 20 / 80) to give 2,2,2-trifluoroethyl (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)phenyl)propanoate (0.965 g). Dichloromethane (8.2 mL) and 4 M hydrochloric acid / MTHP (8.2 mL) were added sequentially to the resulting compound. The mixture was stirred at room temperature and then concentrated under reduced pressure. The residue was suspended and washed with methyl tert-butyl ether and dried in vacuo to give 2,2,2-trifluoroethyl (S)-2-amino-3-(4-(5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)phenyl)propanoate (0.501 g, 0.954 mmol) as a colorless solid.

[0169] Example 1-14: (2,6-Dichloropyridin-4-yl)methyl (S)-2-(methylamino)-5-ureidopentanoate hydrochloride (Compound No. 63)

[0170]

change

[0171] N-(tert-butoxycarbonyl)-L-glutamine (30.0 g, 122 mmol, CAS Registry Number: 13726-85-7), pyridine (210 mL), and dicyclohexylcarbodiimide (22.6 g, 110 mmol, CAS Registry Number: 538-75-0) were added sequentially to a flask equipped with a nitrogen balloon under ice-cooling. After stirring at room temperature for 3 hours, the mixture was filtered and extracted with dichloromethane. The organic layer was washed successively with hydrochloric acid and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 1) to give (S)-2-((tert-butoxycarbonyl)amino)-4-cyanobutanoic acid (23.0 g). To the resulting compound, THF (150 mL) and 60% sodium hydride (8.00 g, 200 mmol, CAS Registry Number: 7646-69-7) were added at 10°C. After stirring the mixture for 30 minutes, iodomethane (110 g, 775 mmol, CAS Registry Number: 74-88-4) was added. The mixture was stirred at room temperature overnight, and then water was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed successively with citric acid and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give (S)-2-((tert-butoxycarbonyl)(methyl)amino)-4-cyanobutanoic acid (13.0 g). To the resulting compound, isopropanol (100 mL), concentrated hydrochloric acid (3 mL), and platinum dioxide (1.00 g, 4.48 mmol, CAS Registry Number: 1314-15-4) were added, in that order. After stirring at room temperature under a hydrogen atmosphere for 6 hours, the mixture was filtered and concentrated under reduced pressure to give (S)-5-amino-2-((tert-butoxycarbonyl)(methyl)amino)pentanoic acid hydrochloride (13.0 g). To the resulting compound, THF (200 mL), water (130 mL), and potassium cyanate (12.9 g, 159 mmol, CAS Registry Number: 590-28-3) were added, in that order. The mixture was stirred at room temperature overnight, and then the reaction was quenched by the addition of hydrochloric acid.The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; acetonitrile / 0.1% formic acid aqueous solution = 10 / 90 → 50 / 50) to give (S)-2-((tert-butoxycarbonyl)(methyl)amino)-5-ureidopentanoic acid (5.03 g). To the resulting compound (3.50 g), DMF (50 mL), 4-(bromomethyl)-2,6-dichloropyridine (2.77 g, 11.5 mmol, CAS Registry Number: 175204-45-2), and DIPEA (2.54 mL, 14.5 mmol) were added in that order. After stirring the mixture at room temperature for 16 hours, the mixture was purified by flash column chromatography (silica gel; methanol / dichloromethane = 1 / 99 → 20 / 80) to give (2,6-dichloropyridin-4-yl)methyl (S)-2-((tert-butoxycarbonyl)(methyl)amino)-5-ureidopentanoate (4.00 g). To the obtained compound (3.00 g), dichloromethane (25 mL) and 4 M hydrochloric acid / CPME (25 mL) were added, successively. After stirring the mixture at room temperature for 2 hours, the mixture was concentrated under reduced pressure. The residue was suspended and washed with diethyl ether to give (2,6-dichloropyridin-4-yl)methyl (S)-2-(methylamino)-5-ureidopentanoate (2.17 g, 5.63 mmol) as a colorless solid.

[0172] Example 1-15: (S)-2-(4-(2-amino-3-((2,6-dichloropyridin-4-yl)methoxy)-3-oxopropyl)piperidin-1-yl)acetic acid dihydrochloride (Compound No. 64)

[0173] [ka]

[0174] To a flask, (S)-2-((tert-butoxycarbonyl)amino)-3-(pyridin-4-yl)propanoic acid (45.0 g, 169 mmol, CAS Registry Number: 37535-57-2), isopropanol (675 mL), 1 M aqueous hydrochloric acid (169 mL), and platinum dioxide (6.52 g, 28.7 mmol, CAS Registry Number: 1314-15-4) were added in this order. After stirring under a hydrogen atmosphere at room temperature for 16 hours, the mixture was filtered and concentrated under reduced pressure to give (S)-2-((tert-butoxycarbonyl)amino)-3-(piperidin-4-yl)propanoic acid hydrochloride (50.0 g). To the resulting compound, THF (1000 mL) and bis(trimethylsilyl)acetamide (115 g, 567 mmol, CAS Registry Number: 10416-59-8) were added in this order under ice-cooling. After stirring for 1 hour under ice-cooling, DIPEA (62.8 mL, 243 mmol) and tert-butyl bromoacetate (47.4 g, 243 mmol, CAS Registry Number: 5292-43-3) were added in this order under ice-cooling. After stirring for 1 hour under ice-cooling and at room temperature for another 1 hour, the mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; methanol / dichloromethane = 1 / 100 → 1 / 1) to give (S)-3-(1-(2-(tert-butoxy)-2-oxoethyl)piperidin-4-yl)-2-((tert-butoxycarbonyl)amino)propanoic acid (55.0 g). To the resulting compound (8.20 g), DMF (80 mL), 4-(bromomethyl)-2,6-dichloropyridine (4.86 g, 20.2 mmol, CAS Registry Number: 175204-45-2), and DIPEA (4.43 mL, 25.5 mmol) were added in this order. The mixture was stirred at room temperature for 2 hours, and then aqueous sodium dihydrogen phosphate was added to quench the reaction. The mixture was extracted with isopropyl acetate, and the organic layer was washed successively with aqueous sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure.The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether=1 / 1) to give (2,6-dichloropyridin-4-yl)methyl (S)-3-(1-(2-(tert-butoxy)-2-oxoethyl)piperidin-4-yl)-2-((tert-butoxycarbonyl)amino)propanoate (7.06 g). To the obtained compound (1.60 g), dichloromethane (15 mL) and 4M hydrochloric acid / MTHP (15 mL) were added in that order. The mixture was stirred at room temperature for 2 hours, and then concentrated under reduced pressure. The residue was suspended and washed with diethyl ether and dried in vacuo to give (S)-2-(4-(2-amino-3-((2,6-dichloropyridin-4-yl)methoxy)-3-oxopropyl)piperidin-1-yl)acetic acid dihydrochloride (1.49 g, 3.22 mmol) as a colorless solid.

[0175] Example 1-16: (2,6-Dichloropyridin-4-yl)methyl (S)-2-amino-3-(1-carbamoylpiperidin-4-yl)propanoate hydrochloride (Compound No. 65)

[0176] [ka]

[0177] To (S)-2-((tert-butoxycarbonyl)amino)-3-(piperidin-4-yl)propanoic acid hydrochloride (80.0 g, 259 mmol) obtained in Example 1-15, THF (600 mL), water (400 mL), and potassium cyanate (36.0 g, 444 mmol, CAS Registry Number: 590-28-3) were added, in that order. The mixture was stirred at room temperature for 16 hours, and then aqueous sodium dihydrogen phosphate was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give (S)-2-((tert-butoxycarbonyl)amino)-3-(1-carbamoylpiperidin-4-yl)propanoic acid (80.0 g). To the resulting compound (15.0 g), DMF (150 mL), 4-(bromomethyl)-2,6-dichloropyridine (10.9 g, 45.2 mmol, CAS Registry Number: 175204-45-2), and DIPEA (9.94 mL, 57.1 mmol) were added, in that order. The mixture was stirred at room temperature for 1 hour, and then water was added to quench the reaction. The mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; methanol / dichloromethane = 1 / 10) to give (2,6-dichloropyridin-4-yl)methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(1-carbamoylpiperidin-4-yl)propanoate (10.7 g). To the resulting compound (3.00 g), dichloromethane (25 mL) and 4M hydrochloric acid / MTHP (25 mL) were added in sequence. The mixture was stirred at room temperature for 30 minutes, and then concentrated under reduced pressure. The residue was suspended in diethyl ether and dried in vacuo to give (2,6-dichloropyridin-4-yl)methyl (S)-2-amino-3-(1-carbamoylpiperidin-4-yl)propanoate hydrochloride (2.90 g, 6.48 mmol) as a colorless solid.

[0178] Example 1-17: (S)-2-(4-((5-amino-6-((2,6-dichloropyridin-4-yl)methoxy)-6-oxohexyl)carbamoyl)piperazin-1-yl)acetic acid dihydrochloride (Compound No. 66)

[0179] [ka]

[0180] To a flask, (tert-butoxycarbonyl)-L-lysine (10.0 g, 40.6 mmol, CAS Registry Number: 13734-28-6), THF (66 mL), and bis(trimethylsilyl)acetamide (18.2 g, 89.3 mmol, CAS Registry Number: 10416-59-8) were added in this order under ice cooling. After stirring the mixture at room temperature for 30 minutes, DIPEA (8.49 mL, 48.7 mmol) and 4-nitrophenyl chloroformate (7.77 g, 38.6 mmol, CAS Registry Number: 7693-46-1) dissolved in THF (17 mL) were added in this order under ice cooling. After stirring at below 10°C for 1.5 hours, the reaction was quenched by the addition of hydrochloric acid. The mixture was extracted with ethyl acetate, and the organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether=1 / 1) to give N 2 -(tert-butoxycarbonyl)-N 6-((4-nitrophenoxy)carbonyl)-L-lysine (10.6 g) was obtained. 1,1-Dimethylethyl 1-piperazine acetate (5.32 g, 26.5 mmol) dissolved in THF (90 mL), DMF (10 mL), and DIPEA (4.84 mL, 27.8 mmol) were added to the obtained compound (10.4 g) in that order under ice-cooling. The mixture was stirred at room temperature for 4 hours, and then aqueous sodium dihydrogen phosphate solution was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; methanol / dichloromethane = 8 / 92) to give N 6 -(4-(2-(tert-butoxy)-2-oxoethyl)piperazine-1-carbonyl)-N 2 4-(tert-Butoxycarbonyl)-L-lysine (8.00 g) was obtained. To the obtained compound, DMF (80 mL), 4-(bromomethyl)-2,6-dichloropyridine (3.87 g, 16.1 mmol, CAS Registry Number: 175204-45-2), and DIPEA (2.63 g, 20.3 mmol) were added in that order. The mixture was stirred at room temperature for 1 hour, and then water was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; methanol / dichloromethane = 4 / 96) to give (2,6-dichloropyridin-4-yl)methyl N 6 -(4-(2-(tert-butoxy)-2-oxoethyl)piperazine-1-carbonyl)-N 2To the resulting compound (5.07 g), dichloromethane (10 mL) and 2,2,2-trifluoroacetic acid (12.5 mL) were added, successively. After stirring at room temperature for 20 hours, the mixture was concentrated under reduced pressure, and acetonitrile and 4M hydrochloric acid / MTHP (25 mL) were added, successively. The mixture was filtered, washed with THF, and dried in vacuo to give (S)-2-(4-((5-amino-6-((2,6-dichloropyridin-4-yl)methoxy)-6-oxohexyl)carbamoyl)piperazin-1-yl)acetic acid dihydrochloride (4.10 g, 8.01 mmol) as a colorless solid.

[0181] Example 1-18: (2,6-Dichloropyridin-4-yl)methyl (S)-2-amino-4-(benzylsulfonyl)butanoate hydrochloride (Compound No. 67)

[0182] [ka]

[0183] To a flask, S-benzyl-N-(tert-butoxycarbonyl)-L-homocysteine ​​(0.325 g, 1.00 mmol, CAS Registry Number: 16947-99-2), methanol (2.0 mL), and magnesium bis(monoperoxyphthalate) hexahydrate (0.594 g, 1.20 mmol, CAS Registry Number: 84665-66-7) were added sequentially under ice-cooling. The mixture was stirred at room temperature, and then hydrochloric acid was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give (S)-4-(benzylsulfonyl)-2-((tert-butoxycarbonyl)amino)butanoic acid (0.357 g). To the resulting compound, DMF (2.0 mL), 4-(bromomethyl)-2,6-dichloropyridine (0.229 g, 0.95 mmol, CAS reg. no.: 175204-45-2), and DIPEA (0.210 mL, 1.20 mmol) were added, in that order. The mixture was stirred at room temperature, and then hydrochloric acid was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / hexane = 0 / 100 → 100 / 0) to give (2,6-dichloropyridin-4-yl)methyl (S)-4-(benzylsulfonyl)-2-((tert-butoxycarbonyl)amino)butanoate (0.474 g). To the resulting compound, dichloromethane (4.6 mL) and 4M hydrochloric acid / MTHP (4.6 mL) were added sequentially. After stirring the mixture at room temperature, the mixture was concentrated under reduced pressure. The residue was suspended in methyl tert-butyl ether and dried in vacuo to give (2,6-dichloropyridin-4-yl)methyl (S)-2-amino-4-(benzylsulfonyl)butanoate hydrochloride (0.384 g, 0.845 mmol) as a colorless solid.

[0184] Example 1-19: (2,6-Dichloropyridin-4-yl)methyl (S)-2-amino-3-(2-carbamoyl-1H-imidazol-4-yl)propanoate dihydrochloride (Compound No. 68)

[0185] [ka]

[0186] To a flask equipped with a nitrogen balloon, 4-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazole (80.0 g, 247 mmol, CAS Registry Number: 518329-44-7) and THF (1400 mL) were added sequentially. After the mixture was stirred at room temperature for 30 minutes, 2M-LDA / THF (148 mL, 296 mmol, CAS Registry Number: 4111-54-0) was added to the mixture at −78° C. After stirring at −78° C. for 30 minutes, the mixture was added to a mixture of ethyl chloroformate (93.7 g, 864 mmol, CAS Registry Number: 541-41-3) and THF (500 mL) at −78° C. After stirring at −78° C. for 30 minutes, water was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 20 / 80 → 50 / 50) to give ethyl 4-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazole-2-carboxylate (80.0 g). 4M-NH3 / MeOH (1600 mL, CAS Registry Number: 7664-41-7) was added to the resulting compound (80.0 g). After stirring at 60 °C for 2.5 hours, the mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 20 / 80 → 50 / 50) to give 4-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazole-2-carboxamide (52.0 g). Zinc (23.5 g, 359 mmol, CAS Registry Number: 7440-66-6), DMF (880 mL), and iodine (9.12 g, 35.9 mmol, CAS Registry Number: 7553-56-2) were added sequentially to a flask equipped with a nitrogen balloon. After stirring at 60°C for 30 minutes, (S)-methyl 2-(tert-butoxycarbonylamino)-3-iodopropanoate (78.9 g, 240 mmol, CAS Registry Number: 93267-04-0) was added to the mixture, which was then stirred at room temperature for 1.5 hours to prepare the organozinc reagent.2-Dicyclohexylphosphino-2',6'-dimethoxybiphenyl (4.92 g, 12.0 mmol, CAS Registry Number: 657408-07-6), tris(dibenzylideneacetone)dipalladium-chloroform (2.74 g, 3.00 mmol, CAS Registry Number: 52522-40-4), and DMF (40 mL) were added in this order to a flask equipped with a nitrogen balloon, and the mixture was stirred at 60°C for 30 minutes to obtain a palladium complex solution.

[0187] The prepared organozinc reagent was added sequentially to the prepared palladium complex solution and 4-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazole-2-carboxamide (44.0 g) dissolved in DMF (160 mL) at room temperature. After stirring at 60 °C for 1 hour, the mixture was filtered and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 20 / 80 → 40 / 60) to give methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(2-carbamoyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol-4-yl)propanoate (30.0 g). To the resulting compound, isopropanol (900 mL), water (300 mL), calcium chloride (120 g, 1080 mmol, CAS Registry Number: 10043-52-4), and lithium hydroxide monohydrate (11.4 g, 271 mmol, CAS Registry Number: 1310-66-3) were added in this order under ice-cooling. The mixture was stirred at room temperature for 2 hours, and then sodium dihydrogen phosphate was added to quench the reaction. The mixture was filtered and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 40 / 60 → 60 / 40) to give (S)-2-((tert-butoxycarbonyl)amino)-3-(2-carbamoyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol-4-yl)propanoic acid (20.0 g). To the resulting compound (20.0 g), DMF (400 mL), 4-(bromomethyl)-2,6-dichloropyridine (10.7 g, 44.3 mmol, CAS registration number: 175204-45-2), and DIPEA (7.24 g, 56.0 mmol) were added in that order. The mixture was stirred at room temperature for 2 hours, and then water was added to quench the reaction.The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; acetonitrile / 0.1% formic acid aqueous solution = 40 / 60 → 60 / 40) to give (2,6-dichloropyridin-4-yl)methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(2-carbamoyl-1-((2-(trimethylsilyl)methoxy)))-1H-imidazol-4-yl)propanoate (10.7 g). Dichloromethane (32 mL) and 4M hydrochloric acid / MTHP (32 mL) were added sequentially to the resulting compound (5.01 g). The mixture was stirred at room temperature for 3 days, and then concentrated under reduced pressure. The residue was suspended and washed with diethyl ether and dried in vacuo. The residue was purified by flash column chromatography (silica gel; acetonitrile / water = 2 / 98 → 15 / 85), and water and acetonitrile were added and the mixture was lyophilized to give (2,6-dichloropyridin-4-yl)methyl (S)-2-amino-3-(6-aminopyridin-3-yl)propanoate dihydrochloride (0.368 g, 0.854 mmol) as a colorless solid.

[0188] Example 1-20: (2,6-dichloropyridin-4-yl)methyl N 5 -(4-Aminobutyl)-L-glutamate dihydrochloride (Compound No. 69)

[0189] [ka]

[0190] Add N to the flask. 2 -(((9H-fluoren-9-yl)methoxy)carbonyl)-N 5-(4-((tert-butoxycarbonyl)amino)butyl)-L-glutamine (2.70 g, 5.00 mmol, purchased from Amatek Chemical Co., Ltd. (AS02649)), acetonitrile (10 mL), and triethylamine (2.53 g, 25.0 mmol) were added sequentially at room temperature. After stirring at 60°C for 1 hour, the mixture was concentrated under reduced pressure. Diethyl ether was added to the residue, filtered, washed with diethyl ether, dried in vacuo, and then purified with N 5 -(4-((tert-butoxycarbonyl)amino)butyl)-L-glutamine was obtained. 1,4-Dioxane (15 mL), water (10 mL), sodium carbonate (1.59 g, 15.0 mmol), and di-tert-butyl dicarbonate (1.31 g, 6.00 mmol, CAS Registry Number: 24424-99-5) were added to the obtained compound in this order at room temperature. After stirring the mixture at room temperature, the mixture was concentrated under reduced pressure and N 2 -(tert-butoxycarbonyl)-N 5 To the resulting compound, DMF (10 mL), DIPEA (0.775 g, 6.00 mmol), and 4-(bromomethyl)-2,6-dichloropyridine (1.14 g, 4.75 mmol, CAS registration number: 175204-45-2) were added in that order. The mixture was stirred at room temperature, and then water was added to quench the reaction. The mixture was extracted with isopropyl acetate, and the organic layer was washed with water and saturated brine in that order, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / heptane = 0 / 100 → 75 / 25) to give (2,6-dichloropyridin-4-yl)methyl N 2 -(tert-butoxycarbonyl)-N 5To the resulting compound, dichloromethane (25 mL) and 4M hydrochloric acid / MTHP (25 mL) were added in that order. The mixture was stirred at room temperature and then concentrated under reduced pressure. The residue was suspended and washed with diethyl ether, and (2,6-dichloropyridin-4-yl)methyl N-(4-((tert-butoxycarbonyl)amino)butyl)-L-glutamate was obtained. 5 -(4-aminobutyl)-L-glutamate dihydrochloride (1.12 g, 2.49 mmol) was obtained as a colorless solid.

[0191] Example 1-21: (2,6-Dichloropyridin-4-yl)methyl O-isobutyl-L-homoserinate hydrochloride (Compound No. 70)

[0192] [ka]

[0193] To a flask were added (S)-4-(tert-butoxy)-3-((tert-butoxycarbonyl)amino)-4-oxobutanoic acid (50.0 g, 173 mmol, CAS Registry Number: 34582-32-6), THF (500 mL), 4-methylmorpholine (22.7 g, 224 mmol, CAS Registry Number; 109-02-4), and isopropyl chloroformate (23.1 g, 188 mmol, CAS Registry Number: 108-23-6) in that order at −10° C. After stirring at −10° C. for 30 minutes, sodium borohydride (13.1 g, 346 mmol, CAS Registry Number: 16940-66-2) was added to the mixture at −10° C. After stirring at -10°C for 30 minutes, sodium borohydride (13.1 g, 346 mmol) dissolved in water (150 mL) was added at -10°C. After stirring at -10°C for 2 hours, aqueous ammonium chloride solution was added to the mixture to quench the reaction. The mixture was extracted with ethyl acetate, washed with aqueous ammonium chloride and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain tert-butyl (tert-butoxycarbonyl)-L-homoserinate (45.0 g). To thionyl chloride (56.2 g, 472 mmol, CAS Registry Number: 7719-09-7) dissolved in acetonitrile (850 mL), tert-butyl (tert-butoxycarbonyl)-L-homoserinate (50.0 g) dissolved in acetonitrile (150 mL) was added at -40°C. After stirring at −40° C. for 15 minutes, pyridine (86.2 g, 1090 mmol) was added to the mixture at −40° C. After stirring at 0° C. for 20 minutes, water was added to the mixture to quench the reaction. The mixture was extracted with dichloromethane, washed with 1 M aqueous hydrochloric acid and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether=1 / 10) to give di-tert-butyl (4S)-1,2,3-oxathiazinane-3,4-dicarboxylate 2-oxide (30.0 g).To the resulting compound, dimethyl carbonate (300 mL) and ruthenium trichloride hydrate (0.21 g, 0.933 mmol, CAS number: 14898-67-0) dissolved in water and sodium periodate (59.9 g, 280 mmol, CAS number: 7790-28-5) were added sequentially under ice-cooling. After stirring at room temperature for 3 hours, the mixture was filtered, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 10) to give di-tert-butyl (S)-1,2,3-oxathiazinane-3,4-dicarboxylate 2,2-dioxide (25.0 g). To the resulting compound (14.5 g), sodium dihydrogen phosphate (20.6 g, 172 mmol, CAS Registry Number: 7558-79-4) and 2-methylpropan-1-ol (79.6 g, 1070 mmol, CAS Registry Number: 78-83-1) were added at room temperature. After stirring at 55 °C for 3 hours, the mixture was concentrated under reduced pressure. The mixture was dissolved in ethyl acetate, washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give tert-butyl N-(tert-butoxycarbonyl)-O-isobutyl-L-homoserinate (13.6 g). To the resulting compound, 1,4-dioxane (35 mL) and 4 M aqueous hydrochloric acid solution (105 mL) were added sequentially at room temperature. After stirring at 85 °C for 3 hours, the mixture was concentrated under reduced pressure to give O-isobutyl-L-homoserine hydrochloride (8.60 g). To the resulting compound, 1,4-dioxane (90 mL), water (60 mL), sodium bicarbonate (13.6 g, 162 mmol, CAS Registry Number: 144-55-8), and di-tert-butyl dicarbonate (10.6 g, 48.6 mmol, CAS Registry Number: 24424-99-5) were added in this order under ice-cooling. The mixture was stirred at room temperature for 5 hours, and then quenched by the addition of 1M aqueous hydrochloric acid. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure.The residue was purified by flash column chromatography (silica gel; methanol / dichloromethane = 1 / 10) to give N-(tert-butoxycarbonyl)-O-isobutyl-L-homoserine (10.4 g). To the resulting compound, DMF (100 mL), DIPEA (5.86 g, 45.3 mmol), and 4-(bromomethyl)-2,6-dichloropyridine (8.64 g, 35.9 mmol, CAS Registry Number: 175204-45-2) were added sequentially. The mixture was stirred at room temperature for 1 hour, and then quenched by the addition of aqueous sodium dihydrogen phosphate. The mixture was extracted with ethyl acetate, and the organic layer was washed successively with aqueous sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 5) to give (2,6-dichloropyridin-4-yl)methyl N-(tert-butoxycarbonyl)-O-isobutyl-L-homoserinate (9.95 g). To the resulting compound (4.98 g), dichloromethane (42 mL) and 4M hydrochloric acid / MTHP (43 mL) were added, successively. The mixture was stirred at room temperature for 17 hours, and then concentrated under reduced pressure. The residue was suspended and washed with diethyl ether to give (2,6-dichloropyridin-4-yl)methyl O-isobutyl-L-homoserinate hydrochloride (3.11 g, 8.37 mmol) as a colorless solid.

[0194] Example 1-22: (2,6-dichloropyridin-4-yl)methyl N 2 ,N 5 ,N 5 -Trimethyl-L-glutamate hydrochloride (Compound No. 71)

[0195] [ka]

[0196] To a flask, ((benzyloxy)carbonyl)-L-glutamic acid (50.0 g, 178 mmol, CAS Registry Number: 1155-62-0), paraformaldehyde (16.0 g, 533 mmol, CAS Registry Number: 30525-89-4), 10-camphorsulfonic acid (8.25 g, 35.6 mmol, CAS Registry Number: 3144-16-9), and toluene (800 mL) were added in this order. After stirring at 80 °C overnight, ethyl acetate was added to the mixture, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; methanol / dichloromethane = 1 / 10) to give (S)-3-(3-((benzyloxy)carbonyl)-5-oxooxazolidin-4-yl)propanoic acid (50.0 g). To the resulting compound (20.0 g), DMF (300 mL), dimethylamine hydrochloride (8.34 g, 102 mmol, CAS Registry Number: 506-59-2), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (51.9 g, 136 mmol, CAS Registry Number: 148893-10-1), and DIPEA (35.3 g, 273 mmol) were added sequentially at room temperature. The mixture was stirred overnight at room temperature, and then ethyl acetate was added to the mixture. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether=1 / 1) to give benzyl (S)-4-(3-(dimethylamino)-3-oxopropyl)-5-oxooxazolidine-3-carboxylate (20.0 g). To the obtained compound (6.50 g), dichloromethane (30 mL), triisopropylsilane (10.5 g, 66.5 mmol, CAS registration number: 6485-79-6), and 2,2,2-trifluoroacetic acid (30.0 mL) were added in this order at room temperature. The mixture was stirred at room temperature overnight, and then concentrated under reduced pressure. 2 -((benzyloxy)carbonyl)-N2 ,N 5 ,N 5 To the resulting compound, methanol (170 mL) and palladium on carbon (1.77 g, CAS Registry Number: 7440-05-3) were added in that order. After stirring overnight at room temperature under a hydrogen atmosphere, the mixture was filtered, extracted with methanol, and concentrated under reduced pressure. 2 ,N 5 ,N 5 1,4-Dioxane (100 mL) and water (100 mL) were added to the resulting compound. After stirring the mixture at room temperature, sodium carbonate (21.4 g, 202 mmol, CAS Registry Number: 497-19-8) and di-tert-butyl dicarbonate (33.1 g, 151 mmol, CAS Registry Number: 24424-99-5) were added in this order under ice-cooling. The mixture was stirred overnight at room temperature, and then hydrochloric acid was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then extracted with N 2 -(tert-butoxycarbonyl)-N 2 ,N 5 ,N 5 To the resulting compound, DMF (100 mL), 4-(bromomethyl)-2,6-dichloropyridine (7.62 g, 31.6 mmol, CAS registration number: 175204-45-2), and DIPEA (5.16 g, 40.0 mmol) were added in that order. The mixture was stirred at room temperature for 1 hour, and then water was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 1) to give (2,6-dichloropyridin-4-yl)methyl N 2 -(tert-butoxycarbonyl)-N 2 ,N 5 ,N 5To the resulting compound (4.48 g), dichloromethane (25 mL) and 4M hydrochloric acid / MTHP (25 mL) were added in that order. The mixture was stirred at room temperature for 17 hours. After the mixture was evaporated, the mixture was concentrated under reduced pressure. The residue was suspended in methyl tert-butyl ether. The resulting mixture was washed with water, dried in vacuo, and purified to give (2,6-dichloropyridin-4-yl)methyl N 2 ,N 5 ,N 5 -trimethyl-L-glutamate hydrochloride (3.54 g, 9.21 mmol) was obtained as a colorless solid.

[0197] Example 1-23: (2,6-Dichloropyridin-4-yl)methyl (S)-3-(5-hydroxypyridin-3-yl)-2-(methylamino)propanoate dihydrochloride (Compound No. 72)

[0198] [ka]

[0199] N-(((9H-fluoren-9-yl)methoxy)carbonyl)-N-methyl-L-serine (150 g, 439 mmol, CAS Registry Number: 291311-48-3), methanol (1500 mL), and concentrated sulfuric acid (50 mL) were added sequentially to a flask equipped with a nitrogen balloon. The mixture was stirred at 60°C and then concentrated under reduced pressure. Ethyl acetate was added to the mixture, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain methyl N-(((9H-fluoren-9-yl)methoxy)carbonyl)-N-methyl-L-serinate (120 g). To the resulting compound (71.0 g), dichloromethane (500 mL), triphenylphosphine (78.6 g, 300 mmol, CAS Registry Number: 603-35-0), imidazole (20.4 g, 300 mmol, CAS Registry Number: 288-32-4), and iodine (76.1 g, 300 mmol, CAS Registry Number: 7553-56-2) were added sequentially under ice cooling. The mixture was stirred at room temperature for 30 minutes, and then dichloromethane was added to the mixture. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 1) to give methyl (R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)(methyl)amino)-3-iodopropanoate (70.0 g). Zinc (19.0 g, 290 mmol, CAS Registry Number: 7440-66-6), DMF (400 mL), and iodine (14.7 g, 58.0 mmol, CAS Registry Number: 7553-56-2) were added sequentially to a flask equipped with a nitrogen balloon. After stirring the mixture at room temperature for 2 minutes, methyl (R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)(methyl)amino)-3-iodopropanoate (45.0 g) was added to the mixture.After stirring the mixture at room temperature for 30 minutes, 5-bromopyridin-3-ol (20.2 g, 116 mmol, CAS Registry Number: 74115-13-2), 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (1.98 g, 4.83 mmol, CAS Registry Number: 657408-07-6), and tris(dibenzylideneacetone)dipalladium-chloroform (2.21 g, 2.42 mmol, CAS Registry Number: 52522-40-4) were added sequentially at room temperature. After stirring at 50°C for 3 hours, water was added to the mixture, which was then filtered and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether=1 / 3) to give methyl (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)(methyl)amino)-3-(5-hydroxypyridin-3-yl)propanoate (18.0 g).

[0200] To the resulting compound (10.0 g), isopropanol (120 mL), water (40 mL), calcium chloride (41.1 g, 370 mmol, CAS Registry Number: 10043-52-4), and lithium hydroxide monohydrate (3.88 g, 92.5 mmol, CAS Registry Number: 1310-66-3) were added sequentially under ice cooling. The mixture was stirred at room temperature overnight, and then sodium dihydrogen phosphate was added to quench the reaction. The mixture was filtered and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 5) to give (S)-2-(((9H-fluoren-9-yl)methoxy)carbonyl)(methyl)amino)-3-(5-hydroxypyridin-3-yl)propanoic acid (8.00 g). To the resulting compound (6.7 g), THF (20 mL) and diethylamine (3.51 g, 48.0 mmol, CAS Registry Number: 109-89-7) were added in that order. After stirring the mixture at room temperature for 1 hour, the mixture was concentrated under reduced pressure to give (S)-3-(5-hydroxypyridin-3-yl)-2-(methylamino)propanoic acid (3.14 g). To the resulting compound, THF (30 mL), di-tert-butyl dicarbonate (7.68 g, 35.2 mmol, CAS Registry Number: 24424-99-5), and DIPEA (4.13 g, 31.2 mmol) were added in that order under ice cooling. The mixture was stirred at room temperature overnight, and then concentrated under reduced pressure to give (S)-2-((tert-butoxycarbonyl)(methyl)amino)-3-(5-((tert-butoxycarbonyl)oxy)pyridin-3-yl)propanoic acid (6.00 g). To the resulting compound were added DMF (30 mL), 4-(bromomethyl)-2,6-dichloropyridine (3.45 g, 14.3 mmol, CAS registration number: 175204-45-2), and DIPEA (2.35 g, 18.2 mmol), in that order. The mixture was stirred at room temperature for 1 hour, and then an aqueous solution of sodium dihydrogen phosphate was added to quench the reaction.The mixture was extracted with ethyl acetate, and the organic layer was washed successively with aqueous sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 4) to give (2,6-dichloropyridin-4-yl)methyl (S)-2-((tert-butoxycarbonyl)(methyl)amino)-3-(5-((tert-butoxycarbonyl)oxy)pyridin-3-yl)propanoic acid (4.95 g). To the resulting compound (2.62 g), dichloromethane (21.3 mL) and 4M hydrochloric acid / MTHP (21.3 mL) were added, successively. The mixture was stirred at room temperature for 4 hours, and then concentrated under reduced pressure. The residue was suspended and washed with diethyl ether and dried in vacuo to give (2,6-dichloropyridin-4-yl)methyl (S)-3-(5-hydroxypyridin-3-yl)-2-(methylamino)propanoate dihydrochloride (1.99 g, 4.64 mmol) as a light brown solid.

[0201] Example 1-24: (2,6-Dichloropyridin-4-yl)methyl (S)-3-(6-aminopyridin-3-yl)-2-(methylamino)propanoate dihydrochloride (Compound No. 73)

[0202] [ka]

[0203] Zinc (22.4 g, 343 mmol, CAS Registry Number: 7440-66-6), DMF (450 mL), and iodine (17.4 g, 68.6 mmol, CAS Registry Number: 7553-56-2) were sequentially added to a flask equipped with a nitrogen balloon. After stirring the mixture at room temperature for 10 minutes, methyl (R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)(methyl)amino)-3-iodopropanoate (53.2 g) obtained from Example 1-23 was added to the mixture. After stirring at room temperature for 1 hour, 5-bromopyridin-2-amine (20.0 g, 116 mmol, CAS Registry Number: 1072-97-5), 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (9.40 g, 22.9 mmol, CAS Registry Number: 657408-07-6), and tris(dibenzylideneacetone)dipalladium-chloroform (5.80 g, 5.72 mmol, CAS Registry Number: 52522-40-4) were added sequentially at room temperature. After stirring at 50°C for 16 hours, water was added to the mixture, which was then filtered and extracted with ethyl acetate. The organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether=2 / 1) to give methyl ((S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)(methyl)amino)-3-(6-aminopyridin-3-yl)propanoate (30.0 g). To the obtained compound, tert-butanol (600 mL), di-tert-butyl dicarbonate (18.2 g, 83.4 mmol, CAS Registry Number: 24424-99-5), and sodium iodide (12.5 g, 83.4 mmol, CAS Registry Number: 7681-82-5) were added, in that order. The mixture was stirred at room temperature for 16 hours, and then concentrated under reduced pressure.The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 4) to give methyl (S)-2-(((9H-fluoren-9-yl)methoxy)carbonyl)(methyl)amino)-3-(6-((tert-butoxycarbonyl)amino)pyridin-3-yl)propanoate (20.0 g). To the resulting compound, isopropanol (360 mL), calcium chloride (66.8 g, 602 mmol, CAS Registry Number: 10043-52-4), water (120 mL), and lithium hydroxide monohydrate (3.61 g, 151 mmol, CAS Registry Number: 1310-66-3) were added in this order under ice cooling. The mixture was stirred at room temperature for 16 hours, and then hydrochloric acid was added to quench the reaction. The mixture was filtered and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give (S)-2-(((9H-fluoren-9-yl)methoxy)carbonyl)(methyl)amino)-3-(6-((tert-butoxycarbonyl)amino)pyridin-3-yl)propanoic acid. THF (50 mL) and diethylamine (5.50 g, 75.2 mmol, CAS Registry Number: 109-89-7) were added sequentially to the obtained compound. The mixture was stirred at room temperature for 5 hours, and then concentrated under reduced pressure to give (S)-3-(6-((tert-butoxycarbonyl)amino)pyridin-3-yl)-2-(methylamino)propanoic acid.

[0204] To the resulting compound, THF, TEA (11.4 g, 75.2 mmol), and di-tert-butyl dicarbonate (9.86 g, 45.2 mmol, CAS Registry Number: 24424-99-5) were added in that order. The mixture was stirred at room temperature for 16 hours, and then concentrated under reduced pressure to give (S)-2-((tert-butoxycarbonyl)(methyl)amino)-3-(6-((tert-butoxycarbonyl)amino)pyridin-3-yl)propanoic acid. To the resulting compound, DMF (200 mL), 4-(bromomethyl)-2,6-dichloropyridine (10.8 g, 44.9 mmol, CAS Registry Number: 175204-45-2), and DIPEA (7.33 g, 56.7 mmol) were added in that order. The mixture was stirred at room temperature for 1 hour, and then aqueous sodium dihydrogen phosphate was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed successively with aqueous sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 5) to give (2,6-dichloropyridin-4-yl)methyl (S)-2-((tert-butoxycarbonyl)(methyl)amino)-3-(6-((tert-butoxycarbonyl)amino)pyridin-3-yl)propanoate (11.3 g). To the resulting compound (4.44 g), dichloromethane (20 mL) and 4M hydrochloric acid / MTHP (20 mL) were added, successively. The mixture was stirred at room temperature, and then concentrated under reduced pressure. The residue was suspended and washed with heptane, dried in vacuo, dissolved in water and acetonitrile, and then lyophilized to give (2,6-dichloropyridin-4-yl)methyl (S)-3-(6-aminopyridin-3-yl)-2-(methylamino)propanoate dihydrochloride (3.33 g, 7.77 mmol) as a colorless solid.

[0205] Example 1-25: (S)-2-(4-((6-((2,6-dichloropyridin-4-yl)methyl)-5-(methylamino)-6-oxohexyl)carbamoyl)piperazin-1-yl)acetic acid dihydrochloride (Compound No. 74)

[0206] [ka]

[0207] In a flask equipped with a nitrogen balloon, add N 2 -(tert-butoxycarbonyl)-N 6 Diazo-L-lysine (50.0 g, 183 mmol, CAS Registry Number: 846549-33-5), THF (500 mL), and 60% sodium hydride (18.4 g, 459 mmol, CAS Registry Number: 7646-69-7) were added in this order under ice-cooling. After stirring for 1 hour under ice-cooling, iodomethane (78.2 g, 551 mmol, CAS Registry Number: 74-88-4) was added to the mixture under ice-cooling. The mixture was stirred overnight at room temperature, and then water was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed successively with aqueous sodium dihydrogen phosphate and saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then extracted with N 2 -(tert-butoxycarbonyl)-N 6 -Diazo-N 2 To the resulting compound, methanol (400 mL) and palladium on carbon (4.00 g, CAS Registry Number: 7440-05-3) were added in that order. After stirring overnight at room temperature under a hydrogen atmosphere, the mixture was filtered, extracted with methanol, concentrated under reduced pressure, and then washed with N 2 -(tert-butoxycarbonyl)-N 2To the resulting compound (30.0 g), methanol (300 mL), bis(trimethylsilyl)acetamide (46.9 g, 230 mmol, CAS Registry Number: 10416-59-8), 1-bromo-2-(2-bromoethoxy)ethane (28.1 g, 121 mmol, CAS Registry Number: 5414-19-7), and DIPEA (44.7 g, 346 mmol) were added in this order at room temperature. After stirring at 60 °C for 24 hours, the mixture was concentrated under reduced pressure to give (S)-2-((tert-butoxycarbonyl)(methyl)amino)-6-morpholinohexanoic acid (30.0 g). To the resulting compound, DMF (300 mL), 4-(bromomethyl)-2,6-dichloropyridine (26.3 g, 109 mmol, CAS Registry Number: 175204-45-2), and DIPEA (17.6 g, 136 mmol) were added in that order. The mixture was stirred at room temperature for 2 hours, and then aqueous sodium dihydrogen phosphate was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed successively with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; acetonitrile / 10 mM aqueous ammonium bicarbonate = 20 / 80 → 60 / 40) to give (2,6-dichloropyridin-4-yl)methyl (S)-2-((tert-butoxycarbonyl)(methyl)amino)-6-morpholinohexanoate (9.80 g). To the resulting compound (4.90 g), dichloromethane (25 mL) and 4 M hydrochloric acid / MTHP (25 mL) were added, successively. The mixture was stirred at room temperature and then concentrated under reduced pressure. Methyl tert-butyl ether was added to the residue, which was then filtered, washed with methyl tert-butyl ether, and dried in vacuo to give (2,6-dichloropyridin-4-yl)methyl (S)-2-(methylamino)-6-morpholinohexanoate dihydrochloride (39.1 g, 8.45 mmol) as a colorless solid.

[0208] Example 1-26: (2,6-Dichloropyridin-4-yl)methyl (S)-2-(methylamino)-6-morpholinohexanoate dihydrochloride (Compound No. 75)

[0209] [ka]

[0210] In a flask, add the N 2 -(tert-butoxycarbonyl)-N 2 To the mixture were added 1-methyl-L-lysine (35.0 g), THF (500 mL), and bis(trimethylsilyl)acetamide (60.2 g, 296 mmol, CAS Registry Number: 10416-59-8), DIPEA (20.9 g, 161 mmol), and 4-nitrophenyl chloroformate (25.2 g, 128 mmol, CAS Registry Number: 7693-46-1) in this order under ice-cooling. The mixture was stirred at room temperature for 2 hours, and then hydrochloric acid was added under ice-cooling to quench the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 1) and purified with N 2 -(tert-butoxycarbonyl)-N 2 -methyl-N 6 -((4-nitrophenoxy)carbonyl)-L-lysine (12.0 g) was obtained. To the obtained compound, THF (108 mL), DMF (12 mL), and 1,1-dimethylethyl 1-piperazine acetate (5.99 g, 29.6 mmol) were added in that order under ice-cooling. The mixture was stirred at 30°C overnight, and then aqueous sodium bicarbonate solution was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. 6 -(4-(2-(tert-butoxy)-2-oxoethyl)piperazine-1-carbonyl)-N 2-(tert-butoxycarbonyl)-N 2 N-methyl-L-lysine (13.0 g) was obtained. To the obtained compound, DMF (60 mL), 4-(bromomethyl)-2,6-dichloropyridine (6.11 g, 25.4 mmol, CAS Registry Number: 175204-45-2), and DIPEA (4.14 g, 32.1 mmol) were added in that order. The mixture was stirred at room temperature for 1 hour, and then aqueous sodium dihydrogen phosphate was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed successively with aqueous sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; acetonitrile / 0.1% aqueous ammonium bicarbonate = 30 / 70 → 50 / 50) to give (2,6-dichloropyridin-4-yl)methyl N-methyl-L-lysine. 6 -(4-(2-(tert-butoxy)-2-oxoethyl)piperazine-1-carbonyl)-N 2 -(tert-butoxycarbonyl)-N 2 To the resulting compound (3.02 g), dichloromethane (14 mL) and 2,2,2-trifluoroacetic acid (14 mL) were added, successively. After stirring at room temperature for 23 hours, the mixture was concentrated under reduced pressure, and acetonitrile and 4M hydrochloric acid / MTHP were added, successively. The mixture was filtered, washed with diethyl ether, and dried in vacuo to obtain (S)-2-(4-((6-((2,6-dichloropyridin-4-yl)methoxy)-5-(methylamino)-6-oxohexyl)carbamoyl)piperazin-1-yl)acetic acid dihydrochloride (2.18 g, 4.67 mmol) as a pale yellow solid.

[0211] Example 1-27: (2,6-Dichloropyridin-4-yl)methyl (S)-2-amino-3-(2-oxo-1,2-dihydropyridin-4-yl)propanoate hydrochloride (Compound No. 76)

[0212] [ka]

[0213] To a flask equipped with a nitrogen balloon, zinc (20.9 g, 319 mmol, CAS Registry Number: 7440-66-6), DMF (600 mL), and iodine (16.2 g, 63.8 mmol, CAS Registry Number: 7553-56-2) were added in that order. After stirring the mixture at room temperature, (S)-methyl 2-(tert-butoxycarbonylamino)-3-iodopropanoate (42.0 g, 128 mmol, CAS Registry Number: 93267-04-0) was added. After stirring the mixture at room temperature for 60 minutes, 4-bromo-2-methoxypyridine (20.0 g, 106 mmol, CAS Registry Number: 100367-39-3), 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (8.73 g, 21.3 mmol, CAS Registry Number: 657408-07-6), and tris(dibenzylideneacetone)dipalladium-chloroform (5.44 g, 5.32 mmol, CAS Registry Number: 52522-40-4) were sequentially added to the mixture. After stirring at 50°C for 16 hours, water was added to the mixture, which was then filtered and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether=1 / 5) to give methyl (S)-2-(((tert-butoxycarbonyl)amino)-3-(2-methoxypyridin-4-yl)propanoate (25.0 g). 1,4-Dioxane (250 mL) and 48% aqueous hydrogen bromide solution (250 mL) were added to the resulting compound, successively. After stirring at 100° C. overnight, the mixture was concentrated under reduced pressure to give (S)-2-amino-3-(2-oxo-1,2-dihydropyridin-4-yl)propanoic acid hydrobromide. The resulting compound was purified by 1,4-dioxane (300 mL), water (300 mL), DIPEA (49.4 g, 380 mmol), and di-tert-butyl dicarbonate (24.9 g, 114 mmol, CAS Registry Number: 24424-99-5) was added in turn under ice-cooling. After the mixture was stirred at room temperature for 4 hours, the reaction was quenched by adding aqueous hydrochloric acid, and the mixture was concentrated under reduced pressure.The residue was purified by flash column chromatography (silica gel; acetonitrile / 10 mM aqueous ammonium bicarbonate = 10 / 90 → 50 / 50) to give (S)-2-((tert-butoxycarbonyl)amino)-3-(2-oxo-1,2-dihydropyridin-4-yl)propanoic acid (20.0 g). To the resulting compound, DMF (150 mL), DIPEA (11.1 g, 63.8 mmol), and 4-(bromomethyl)-2,6-dichloropyridine (12.2 g, 50.5 mmol, CAS Registry Number: 175204-45-2) were added, in that order. The mixture was stirred at room temperature for 2 hours, and then quenched by the addition of 1 M aqueous hydrochloric acid. The mixture was extracted with ethyl acetate, and the organic layer was washed successively with aqueous sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; methanol / dichloromethane = 1 / 10) to give (2,6-dichloropyridin-4-yl)methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(2-oxo-1,2-dihydropyridin-4-yl)propanoate (10.8 g). Dichloromethane (42 mL) and 4M hydrochloric acid / MTHP (42 mL) were added sequentially to the obtained compound (5.00 g). The mixture was stirred at room temperature for 5 hours and then concentrated under reduced pressure. The residue was suspended and washed with diethyl ether and dried in vacuo to give (2,6-dichloropyridin-4-yl)methyl (S)-2-amino-3-(2-oxo-1,2-dihydropyridin-4-yl)propanoate hydrochloride (4.85 g, 12.81 mmol) as a colorless solid.

[0214] Example 1-28: (2,6-Dichloropyridin-4-yl)methyl (S)-2-amino-4-(2-oxo-1,2-dihydroquinolin-6-yl)butanoate hydrochloride (Compound No. 77)

[0215] [ka]

[0216] To a flask equipped with a nitrogen balloon, zinc (16.5 g, 252 mmol, CAS Registry Number: 7440-66-6), DMF (500 mL), and iodine (6.40 g, 25.2 mmol, CAS Registry Number: 7553-56-2) were added in that order. After stirring the mixture at room temperature, (S)-2-((tert-butoxycarbonyl)amino)-4-iodobutanoate (34.6 g, 101 mmol, CAS Registry Number: 101650-14-0) was added. After stirring the mixture at room temperature for 60 minutes, 6-bromo-2-methoxyquinoline (20.0 g, 84.0 mmol, CAS Registry Number: 99455-05-7), 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (6.90 g, 16.8 mmol, CAS Registry Number: 657408-07-6), and tris(dibenzylideneacetone)dipalladium-chloroform (4.35 g, 4.20 mmol, CAS Registry Number: 52522-40-4) were sequentially added to the mixture. After stirring at 50°C for 3 hours, water was added to the mixture, which was then filtered and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether=1 / 5) to give methyl (S)-2-(((tert-butoxycarbonyl)amino)-4-(2-methoxyquinolin-6-yl)butanoate (25.0 g). 1,4-Dioxane (300 mL) and 48% aqueous hydrogen bromide solution (300 mL) were added to the resulting compound, successively. After stirring at 100° C. overnight, the mixture was concentrated under reduced pressure to give (S)-2-amino-4-(2-oxo-1,2-dihydroquinolin-6-yl)butanoic acid hydrobromide. The resulting compound was dissolved in 1,4-dioxane (200 mL), water (200 mL), DIPEA (34.8 g, 269 mmol), and di-tert-butyl dicarbonate (17.6 g, 80.7 mmol, CAS Registry Number: 24424-99-5) was added under ice-cooling. After the mixture was stirred at room temperature overnight, the reaction was quenched by adding aqueous hydrochloric acid, and the mixture was concentrated under reduced pressure.The residue was purified by flash column chromatography (silica gel; methanol / dichloromethane = 1 / 5) to give (S)-2-((tert-butoxycarbonyl)amino)-4-(2-oxo-1,2-dihydroquinolin-6-yl)butanoic acid (20.0 g). To the resulting compound (12.0 g), DMF (120 mL), DIPEA (5.37 g, 41.6 mmol), and 4-(bromomethyl)-2,6-dichloropyridine (7.93 g, 32.9 mmol, CAS Registry Number: 175204-45-2) were added, in that order. The mixture was stirred at room temperature for 2 hours, and then quenched by the addition of 1M aqueous sodium bicarbonate solution. The mixture was extracted with ethyl acetate, and the organic layer was washed successively with aqueous sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / dichloromethane = 1 / 1) to give (2,6-dichloropyridin-4-yl)methyl (S)-2-((tert-butoxycarbonyl)amino)-4-(2-oxo-1,2-dihydroquinolin-6-yl)butanoate (10.1 g). To the resulting compound (3.66 g), dichloromethane (27 mL) and 4M hydrochloric acid / MTHP (27 mL) were added, successively. The mixture was stirred at room temperature for 16 hours, and then concentrated under reduced pressure. The residue was added diethyl ether, filtered, washed with diethyl ether, and dried in vacuo to give (2,6-dichloropyridin-4-yl)methyl (S)-2-amino-4-(2-oxo-1,2-dihydroquinolin-6-yl)butanoate hydrochloride (3.14 g, 7.09 mmol) as a colorless solid.

[0217] Example 1-29: 2,2,2-trifluoroethyl (S)-2-amino-3-(4-(2-aminopyridin-3-yl)phenyl)propanoate dihydrochloride (Compound No. 78)

[0218] [ka]

[0219] A flask was charged with (S)-3-(4-bromophenyl)-2-(((tert-butoxycarbonyl)amino)propanoic acid (20.0 g, 58.1 mmol, CAS Registry Number: 62129-39-9), 1,4-dioxane (160 mL), water (40 mL), (2-((tert-butoxycarbonyl)amino)pyridin-3-yl)boronic acid (16.6 g, 69.7 mmol, CAS Registry Number: 863753-35-9), potassium carbonate (24.1 g, 174 mmol), and [1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium (3.03 g, 4.65 mmol, CAS Registry Number: 95408-45-0) was added sequentially at room temperature. After stirring at 50°C under a nitrogen atmosphere for 6 hours, the reaction was quenched by adding 1M aqueous hydrochloric acid. The mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; methanol / dichloroethane = 1 / 5) to give (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(2-((tert-butoxycarbonyl)amino)pyridin-3-yl)phenyl)propanoic acid (14.5 g). The resulting compound was dissolved in DMF (150 mL), 2,2,2-trifluoroethyl trifluoromethanesulfonate (6.99 g, 30.1 mmol, CAS Registry Number: 6226-25-1), and DIPEA (4.92 g, 38.0 mmol). (mmol) was added sequentially. The mixture was stirred at room temperature for 3 hours, and then aqueous sodium dihydrogen phosphate was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed with aqueous sodium bicarbonate and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether=1 / 1) to give 2,2,2-trifluoroethyl (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(2-((tert-butoxycarbonyl)amino)pyridin-3-yl)phenyl)propanoate (10.3 g).To the resulting compound (4.32 g), dichloromethane (20 mL) and 4M hydrochloric acid / MTHP (20 mL) were added, successively. The mixture was stirred at room temperature and then concentrated under reduced pressure. The residue was triturated with methyl tert-butyl ether and dried in vacuo to give 2,2,2-trifluoroethyl (S)-2-amino-3-(4-(2-aminopyridin-3-yl)phenyl)propanoate dihydrochloride (3.33 g, 8.10 mmol) as a light beige solid.

[0220] Example 1-30: (2,6-Dichloropyridin-4-yl)methyl (S)-2-amino-3-(2-carbamoyl-1H-imidazol-4-yl)propanoate dihydrochloride (Compound No. 79)

[0221] [ka]

[0222] 2-(1-trityl-1H-imidazol-4-yl)ethan-1-amine (11.7 g, 33.1 mmol, CAS Registry Number: 195053-92-0), acetonitrile (300 mL), DIPEA (8.56 g, 66.2 mmol), and 2-bromobenzyl acetate (8.34 g, 36.4 mmol, CAS Registry Number: 5437-45-6) were added to a flask in this order under ice cooling. The mixture was stirred at room temperature for 5 hours, and then water was added to quench the reaction. The mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; methanol / dichloroethane = 0 / 100 → 7 / 93) to give benzyl (2-(1-trityl-1H-imidazol-4-yl)ethyl)glycinate (13.6 g). Methanol (120 mL) and palladium on carbon (4.08 g, CAS Registry Number: 7440-05-3) were added sequentially to the obtained compound. After stirring under a hydrogen atmosphere at room temperature for 4 hours, the mixture was filtered, extracted with methanol, and concentrated under reduced pressure to give (2-(1-trityl-1H-imidazol-4-yl)ethyl)glycine (11.5 g). To the resulting compound (5.70 g), 1,4-dioxane (30 mL), water (10 mL), sodium bicarbonate (2.33 g, 27.7 mmol), and di-tert-butyl dicarbonate (3.63 g, 16.6 mmol, CAS Registry Number: 24424-99-5) were added sequentially under ice-cooling. The mixture was stirred at room temperature for 2 hours, and then quenched by the addition of aqueous citric acid. The mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; methanol / dichloroethane = 1 / 100 → 1 / 10) to give N-(tert-butoxycarbonyl)-N-(2-(1-trityl-1H-imidazol-4-yl)ethyl)glycine (4.50 g).To the resulting compound, DMF (20 mL), 4-(bromomethyl)-2,6-dichloropyridine (2.01 g, 8.36 mmol, CAS Registry Number: 175204-45-2), and DIPEA (1.36 g, 4.69 mmol) were added in that order. The mixture was stirred at room temperature for 2 hours, and then water was added to quench the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed successively with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 100 → 1 / 1) to give (2,6-dichloropyridin-4-yl)methyl N-(tert-butoxycarbonyl)-N-(2-(1-trityl-1H-imidazol-4-yl)ethyl)glycinate (2.69 g). To the resulting compound (1.50 g), dichloromethane (3.5 mL), triisopropylsilane (1.0 mL), and 2,2,2-trifluoroacetic acid (3.4 mL) were added in this order. After stirring the mixture at room temperature for 2 hours, the mixture was concentrated under reduced pressure, and acetonitrile and 4M hydrochloric acid / MTHP were added in this order. After stirring the mixture at room temperature for 30 minutes, the mixture was filtered, washed with diethyl ether, and dried in vacuo to give (2,6-dichloropyridin-4-yl)methyl (S)-2-amino-3-(2-carbamoyl-1H-imidazol-4-yl)propanoate dihydrochloride (0.487 g, 1.21 mmol) as a gray solid.

[0223] Example 1-31: (2,6-Dichloropyridin-4-yl)methyl (S)-2-amino-3-(3-((3-aminopropyl)carbamoyl)phenyl)propanoate dihydrochloride (Compound No. 80)

[0224] [ka]

[0225] 3-Bromobenzoic acid (10.0 g, 49.7 mmol, CAS Registry Number: 585-76-2), DMF (200 mL), tert-butyl (3-aminopropyl)carbamate (10.4 g, 59.7 mmol, CAS Registry Number: 75178-96-0), 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide hydrochloride (10.5 g, 54.7 mmol, CAS Registry Number: 25952-53-8), and ethyl 2-cyano-2-(hydroxyimino)acetate (7.78 g, 54.7 mmol, CAS Registry Number: 3849-21-6) were added to a flask at room temperature. The mixture was stirred at room temperature for 2 hours, and then water was added to quench the reaction. The mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 1) to give tert-butyl (3-(3-bromobenzamido)propyl)carbamate (15.0 g). Zinc (8.24 g, 126 mmol, CAS Registry Number: 7440-66-6), DMF (300 mL), and iodine (3.20 g, 12.6 mmol, CAS Registry Number: 7553-56-2) were added sequentially to a flask equipped with a nitrogen balloon. The mixture was stirred at room temperature, and then methyl (R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-iodopropanoate (20.8 g, 46.2 mmol, CAS Registry Number: 156017-42-4) and iodine (3.20 g, 12.6 mmol, CAS Registry Number: 7553-56-2) were added to the mixture in that order.After stirring the mixture at room temperature for 30 minutes, the obtained tert-butyl (3-(3-bromobenzamido)propyl)carbamate (15.0 g), 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (3.45 g, 8.40 mmol, CAS Registry Number: 657408-07-6), and tris(dibenzylideneacetone)dipalladium-chloroform (1.92 g, 2.10 mmol, CAS Registry Number: 52522-40-4) were added to the mixture at room temperature in that order. After stirring at 50 °C for 3 hours, the mixture was filtered, ethyl acetate was added, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was suspended and washed with diethyl ether and dried in vacuo to give methyl (S)-2-(((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(3-(((tert-butoxycarbonyl)amino)propyl)carbamoyl)phenyl)propanoate (20.0 g).

[0226] To the resulting compound (23.0 g), isopropyl alcohol (300 mL), water (100 mL), THF (50 mL), calcium chloride (67.9 g, 612 mmol, CAS Registry Number: 10043-52-4), and lithium hydroxide monohydrate (6.42 g, 153 mmol, CAS Registry Number: 1310-66-3) were added in this order under ice-cooling. The mixture was stirred at room temperature, and then aqueous sodium dihydrogen phosphate was added to quench the reaction. The mixture was filtered, extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / petroleum ether = 1 / 1) to give (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(3-(((tert-butoxycarbonyl)amino)propyl)carbamoyl)phenyl)propanoic acid (19.5 g). To the obtained compound (1.20 g), acetonitrile (10 mL) and triethylamine (1.01 g, 10.0 mmol) were added in that order at room temperature. The mixture was stirred at 60°C for 1 hour and then concentrated under reduced pressure. The residue was suspended and washed with diethyl ether and dried in vacuo to give (S)-2-amino-3-(3-(((tert-butoxycarbonyl)amino)propyl)carbamoyl)phenyl)propanoic acid. To the obtained compound were added 1,4-dioxane (6.0 mL), water (4.0 mL), sodium carbonate (0.636 g, 6.00 mmol), and di-tert-butyl dicarbonate (0.524 g, 2.4 mmol, CAS registration number: 24424-99-5) in this order at room temperature.

[0227] The mixture was stirred at room temperature and then concentrated under reduced pressure to give (S)-2-((tert-butoxycarbonyl)amino)-3-(3-((tert-butoxycarbonyl)amino)propyl)phenyl)propanoic acid. To the resulting compound were added DMF (4.0 mL) and 4-(bromomethyl)-2,6-dichloropyridine (0.458 g, 1.90 mmol, CAS registration number: 175204-45-2), in that order. The mixture was stirred at room temperature and then quenched by the addition of water. The mixture was extracted with isopropyl acetate, and the organic layer was washed successively with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; ethyl acetate / heptane = 0 / 100 → 80 / 20) to give (2,6-dichloropyridin-4-yl)methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(3-((tert-butoxycarbonyl)amino)carbamoyl)phenyl)propanoate. Dichloromethane (25 mL) and 4 M hydrochloric acid / MTHP (25 mL) were added sequentially to the obtained compound. After stirring at room temperature, the mixture was concentrated under reduced pressure. The residue was suspended and washed with diethyl ether and dried in vacuo to give (2,6-dichloropyridin-4-yl)methyl (S)-2-amino-3-(3-((3-aminopropyl)carbamoyl)phenyl)propanoate dihydrochloride (0.262 g, 0.525 mmol) as a colorless solid.

[0228] Example 1-32: (2,6-Dichloropyridin-4-yl)methyl (S)-2-amino-4-(3,3-difluoropiperidin-1-yl)butanoate dihydrochloride (Compound No. 81)

[0229] [ka]

[0230] A flask was charged with (S)-4-(tert-butoxy)-3-((tert-butoxycarbonyl)amino)-4-oxobutanoic acid (40.0 g, 138 mmol, CAS Registry Number: 34582-32-6), DMF (800 mL), 3,3-difluoropiperidine hydrochloride (24.0 g, 152 mmol, CAS Registry Number: 496807-97-7), 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide hydrochloride (29.2 g, 152 mmol, CAS Registry Number: 25952-53-8), ethyl 2-cyano-2-(hydroxyimino)acetate (21.6 g, 152 mmol, CAS Registry Number: 3849-21-6), and DIPEA (26.8 g, 207 (mmol) was added sequentially under ice-cooling. The mixture was stirred at room temperature overnight, and then aqueous sodium bicarbonate solution was added to quench the reaction. The mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; methanol / dichloromethane = 1 / 20) to give tert-butyl (S)-2-((tert-butoxycarbonyl)amino)-4-(3,3-difluoropiperidin-1-yl)-4-oxobutanoate (44.0 g). To a solution of 1,1,3,3-tetramethyldisiloxane (120 g, 897 mmol, CAS Registry Number: 3277-26-7) and dodecacarbonyltriruthenium (3.58 g, 5.61 mmol, CAS Registry Number: 15243-33-1) in THF was added tert-butyl (S)-2-((tert-butoxycarbonyl)amino)-4-(3,3-difluoropiperidin-1-yl)-4-oxobutanoate (44.0 g) dissolved in THF (800 mL). After stirring overnight at 40°C under an atmospheric atmosphere, the mixture was concentrated under reduced pressure to give tert-butyl (S)-2-(((tert-butoxycarbonyl)amino)-4-(3,3-difluoropiperidin-1-yl)butanoate. 1,4-Dioxane (300 mL) and 4 M aqueous hydrochloric acid solution (300 mL) were added to the obtained compound in that order at room temperature.After stirring overnight at 100°C, the mixture was concentrated under reduced pressure to give (S)-2-amino-4-(3,3-difluoropiperidin-1-yl)butanoic acid dihydrochloride. 1,4-Dioxane (300 mL), water (300 mL), sodium carbonate (57.5 g, 542 mmol, CAS Registry Number: 497-19-8), and di-tert-butyl dicarbonate (35.5 g, 163 mmol, CAS Registry Number: 24424-99-5) were added to the resulting compound in this order under ice cooling. The mixture was stirred overnight at room temperature, and then aqueous sodium dihydrogen phosphate was added to quench the reaction.

[0231] The mixture was extracted with ethyl acetate, and the organic layer was washed successively with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give (S)-2-((tert-butoxycarbonyl)amino)-4-(3,3-difluoropiperidin-1-yl)butanoic acid (14.0 g). To the resulting compound, DMF (150 mL), DIPEA (6.74 g, 52.1 mmol), and 4-(bromomethyl)-2,6-dichloropyridine (9.94 g, 41.3 mmol, CAS Registry Number: 175204-45-2) were added in that order. The mixture was stirred at room temperature for 1 hour, and then quenched by the addition of 1 M aqueous hydrochloric acid. The mixture was extracted with ethyl acetate, and the organic layer was washed successively with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; acetonitrile / 0.1% formic acid aqueous solution = 5 / 95 → 100 / 0) to give (2,6-dichloropyridin-4-yl)methyl (S)-2-((tert-butoxycarbonyl)amino)-4-(3,3-difluoropiperidin-1-yl)butanoate (7.30 g). To the obtained compound (3.22 g), dichloromethane (20 mL) and 4M-hydrochloric acid / MTHP (20 mL) were added, successively. The mixture was stirred at room temperature for 4 hours and then concentrated under reduced pressure. The residue was suspended and washed with diethyl ether and dried in vacuo to give (2,6-dichloropyridin-4-yl)methyl (S)-2-amino-4-(3,3-difluoropiperidin-1-yl)butanoate dihydrochloride (2.68 g, 5.89 mmol) as a colorless solid.

[0232] Using the corresponding starting amino acid, esterification reagent, and deprotection conditions in the table, Example 1-1 The following compounds were synthesized according to the procedure described above.

[0233] [Table 1-1]

[0234] Table 1-2

[0235] Table 1-3

[0236] Table 1-4

[0237] Table 1-5

[0238] Table 1-6

[0239] Table 1-7

[0240] Table 1-8

[0241] Table 1-9

[0242] Table 1-10

[0243] Table 1-11

[0244] Table 1-12

[0245] Table 1-13

[0246] Table 1-14

[0247] Table 1-15

[0248] Table 1-16

[0249] Table 1-17

[0250] Table 1-18

[0251] Table 1-19

[0252] Table 1-20

[0253] Table 1-21

[0254] Table 1-22

[0255] Table 1-23

[0256] Table 1-24

[0257] Table 1-25

[0258] Table 1-26

[0259] Table 1-27

[0260] Table 1-28

[0261] Table 1-29

[0262] Table 1-30

[0263] Table 1-31

[0264] Table 1-32

[0265] Table 1-33

[0266] Table 1-34

[0267] Table 1-35

[0268] Table 1-36

[0269] Table 1-37

[0270] Table 1-38

[0271] Table 1-39

[0272] Table 1-40

[0273] Table 1-41

[0274] Table 1-42

[0275] Table 1-43

[0276] Table 1-44

[0277] Table 1-45

[0278] Table 1-46

[0279] Table 1-47

[0280] Table 1-48

[0281] Table 1-49

[0282] Table 1-50

[0283] Table 1-51

[0284] Table 1-52

[0285] Table 1-53

[0286] Table 1-54

[0287] Table 1-55

[0288] Table 1-56

[0289] Table 1-57

[0290] Table 1-58

[0291] Table 1-59

[0292] Table 1-60

[0293] Table 1-61

[0294] Table 1-62

[0295] Table 1-63

[0296] Table 1-64

[0297] Table 1-65

[0298] Table 1-66

[0299] Table 1-67

[0300] Table 1-68

[0301] Table 1-69

[0302] Table 1-70

[0303] Table 1-71

[0304] Table 1-72

[0305] Table 1-73

[0306] Table 1-74

[0307] Table 1-75

[0308] Table 1-76

[0309] Table 1-77

[0310] Table 1-78

[0311] Table 1-79

[0312] Table 1-80

[0313] Table 1-81

[0314] Table 1-82

[0315] Table 1-83

[0316] Table 1-84

[0317] Table 1-85

[0318] Table 1-86

[0319] Table 1-87

[0320] Table 1-88

[0321] Table 1-89

[0322]

Table 1-90

[0323] Table 1-91

[0324] Table 1-92

[0325] Table 1-93

[0326] Table 1-94

[0327] Table 1-95

[0328] Table 1-96

[0329] Table 1-97

[0330] Table 1-98

[0331] Table 1-99

[0332] Table 1-100

[0333] Table 1-101

[0334] Table 1-102

[0335] Table 1-103

[0336] Table 1-104

[0337] Table 1-105

[0338] Table 1-106

[0339] Table 1-107

[0340] Table 1-108

[0341] Table 1-109

[0342]

Table 1-110

[0343] Table 1-111

[0344] Table 1-112

[0345] Table 1-113

[0346] Table 1-114

[0347] Table 1-115

[0348] Table 1-116

[0349] Table 1-117

[0350] Table 1-118

[0351] Table 1-119

[0352]

Table 1-120

[0353] Table 1-121

[0354] Table 1-122

[0355]

Table 1-123

[0356] Table 1-124

[0357] Table 1-125

[0358] Table 1-126

[0359] Table 1-127

[0360] Table 1-128

[0361] Table 1-129

[0362] Table 1-130

[0363] Table 1-131

[0364] Table 1-132

[0365] Table 1-133

[0366] Table 1-134

[0367] Table 1-135

[0368] Table 1-136

[0369] Table 1-137

[0370] Table 1-138

[0371] Table 1-139

[0372] Table 1-140

[0373] Table 1-141

[0374] Table 1-142

[0375] Table 1-143

[0376] [Table 1-144]

[0377] [Table 1-145]

[0378] [Table 1-146]

[0379] [Table 1-147]

[0380] The compounds shown below were synthesized according to the procedures in Examples 1-2 using the corresponding starting amino acids in the table.

[0381] [Table 2-1]

[0382] [Table 2-2]

[0383] [Table 2-3]

[0384] [Table 2-4]

[0385] [Table 2-5]

[0386] [Table 2-6]

[0387] [Table 2-7]

[0388] [Table 2-8]

[0389] The compounds shown below were synthesized according to the procedures of Examples 1-3 using the corresponding starting amino acids and esterification reagents in the table.

[0390] [Table 3-1]

[0391] [Table 3-2]

[0392] [Table 3-3]

[0393] The compounds shown below were synthesized according to the procedures of Examples 1-4 using the corresponding starting amino acids and esterification reagents in the table.

[0394] [Table 4-1]

[0395] [Table 4-2]

[0396] The compounds shown below were synthesized according to the procedures of Examples 1-5 using the corresponding starting amino acids and esterification reagents in the table.

[0397] [Table 5-1]

[0398] [Table 5-2]

[0399] [Table 5-3]

[0400] The compounds shown below were synthesized according to the procedures in Examples 1-6 using the corresponding starting materials in the table.

[0401] [Table 6-1]

[0402] [Table 6-2]

[0403] [Table 6-3]

[0404] [Table 6-4]

[0405] [Table 6-5]

[0406] [Table 6-6]

[0407] The compounds shown below were synthesized according to the procedures of Examples 1-7 using the corresponding starting compounds in the table.

[0408] [Table 7-1]

[0409] [Table 7-2]

[0410] [Table 7-3]

[0411] [Table 7-4]

[0412] [Table 7-5]

[0413] [Table 7-6]

[0414] [Table 7-7]

[0415] [Table 7-8-1] [Table 7-8-2]

[0416] [Table 7-9]

[0417] The compounds shown below were synthesized according to the procedures of Examples 1-8 using the corresponding starting compounds in the table.

[0418] [Table 8-1] [Table 8-2]

[0419] The compounds shown below were synthesized according to the procedures of Examples 1-9 using the corresponding starting compounds in the table.

[0420] [Table 9-1]

[0421] [Table 9-2]

[0422] The compounds shown below were synthesized according to the procedures of Examples 1-10 using the corresponding starting compounds in the table.

[0423] [Table 10]

[0424] The compounds shown below were synthesized according to the procedures of Examples 1-11 using the corresponding starting compounds in the table.

[0425] [Table 11-1]

[0426] [Table 11-2]

[0427] Following the procedures of Examples 1-12, the compounds shown below were synthesized.

[0428] [Table 12]

[0429] Following the procedures of Examples 1-13, the compounds shown below were synthesized.

[0430] [Table 13]

[0431] Following the procedures of Examples 1-14, the compounds shown below were synthesized.

[0432] [Table 14-1] [Table 14-2]

[0433] Following the procedures of Examples 1-15, the compounds shown below were synthesized.

[0434] [Table 15]

[0435] Following the procedures of Examples 1-16, the compounds shown below were synthesized. [Table 16]

[0436] Following the procedures of Examples 1-17, the compounds shown below were synthesized. [Table 17]

[0437] Following the procedures of Examples 1-18, the compounds shown below were synthesized.

[0438] [Table 18]

[0439] Following the procedures of Examples 1-19, the compounds shown below were synthesized.

[0440] [Table 19]

[0441] The compounds shown below were synthesized according to the procedures of Examples 1-20 using the corresponding starting amino acids in the table.

[0442] [Table 20-1]

[0443] [Table 20-2]

[0444] Following the procedures of Examples 1-21, the compounds shown below were synthesized.

[0445] [Table 21]

[0446] The compounds shown below were synthesized according to the procedures of Examples 1-22 using the corresponding starting compounds in the table.

[0447] [Table 22-1]

[0448] [Table 22-2-1] [Table 22-2-2]

[0449] The compounds shown below were synthesized according to the procedures of Examples 1-23 using the corresponding starting compounds in the table.

[0450] [Table 23-1]

[0451] [Table 23-2]

[0452] Following the procedures of Examples 1-24, the compounds shown below were synthesized.

[0453] [Table 24]

[0454] Following the procedures of Examples 1-25, the compounds shown below were synthesized.

[0455] [Table 25]

[0456] Following the procedures of Examples 1-26, the compounds shown below were synthesized.

[0457] [Table 26]

[0458] The compounds shown below were synthesized according to the procedures of Examples 1-27 using the corresponding starting compounds in the table.

[0459] [Table 27-1]

[0460] [Table 27-2]

[0461] [Table 27-3]

[0462] Following the procedures of Examples 1-28, the compounds shown below were synthesized.

[0463] [Table 28]

[0464] Following the procedures of Examples 1-29, the compounds shown below were synthesized. [Table 29-1] [Table 29-2]

[0465] Following the procedures of Examples 1-30, the compounds shown below were synthesized.

[0466] [Table 30]

[0467] The compounds shown below were synthesized according to the procedures in Examples 1-31 using the corresponding starting amino acids in the table.

[0468] [Table 31]

[0469] Following the procedures of Examples 1-32, the compounds shown below were synthesized.

[0470] [Table 32]

[0471] Example 2: Confirmation of tRNA aminoacylation and measurement of aminoacylation efficiency In this embodiment, Flexizyme was used to perform aminoacylation of tRNA using the activated ester synthesized in Example 1 as a substrate. To simply confirm acylation with various amino acids, the initiator tRNA (tRNA Met Instead of α-aminohelix, a short-chain analogue of α-aminohelix, the acylation reaction was performed using microhelix. The resulting solution was analyzed by polyacrylamide gel electrophoresis under acidic conditions to confirm the efficiency of aminoacylation. The band derived from microhelix exhibits slower mobility upon aminoacylation. The efficiency of aminoacylation can be determined by comparing the intensity of the microhelix band and the acylated microhelix band.

[0472] Example 2-1: Measurement of tRNA aminoacylation efficiency of (2,6-dichloropyridin-4-yl)methyl-L-isoleucinate (Compound No. 21) and (2,6-dichloropyridin-4-yl)methyl glycinate (Compound No. 99) The acylation reaction was carried out overnight on ice in 50 mM Hepes-K buffer (pH 7.5), 50 mM MgCl, 25 μM tRNA analog (microhelix), and 5 mM substrate in 20% DMSO. The nucleotide sequence of dFx is shown in SEQ ID NO: 3.

[0473] The detailed procedure was as follows: 1 μL of 500 mM Hepes-K buffer (pH 7.5), 1 μL of 250 μM Flexizyme (dFx), and 1 μL of 250 μM tRNA analog were added to 3 μL of ultrapure water, heated at 95°C for 2 minutes, and cooled at room temperature for 5 minutes. 2 μL of 250 mM MgCl2 was added and the mixture was incubated on ice for 1 minute. The acylation reaction of the tRNA analog was initiated by adding 1.5 μL of DMSO and 0.5 μL of 100 mM substrate, and the mixture was incubated on ice overnight. The reaction was stopped by adding 1 μL of 3 M sodium acetate, pH 5.2. This solution was analyzed by 20% denaturing PAGE (50 mM sodium acetate, 6 M urea) under acidic conditions.

[0474] The results of acylation using (2,6-dichloropyridin-4-yl)methyl L-isoleucinate (Compound No. 21) or (2,6-dichloropyridin-4-yl)methyl glycinate (Compound No. 99) as the substrate are shown in Figures 1-1 and 1-2. These results confirmed that amino acid derivatives were efficiently acylated by Flexizyme.

[0475] Example 2-2: 2,2,2-trifluoroethyl(S)-2-amino-4-phenyl Measurement of the aminoacylation efficiency of tRNA with butanoate (Compound No. 47) and 2,2,2-trifluoroethyl (S)-2-amino-3-(1H-pyrrolo[2,3-b]pyridin-3-yl)propanoate (Compound No. 81) The acylation reaction was carried out overnight on ice in 50 mM Hepes-K buffer (pH 7.5), 600 mM MgCl, 25 μM flexizyme (eFx), 25 μM tRNA analog (microhelix), and 5 mM substrate in 20% DMSO. The nucleic acid sequence of eFx is shown in SEQ ID NO: 2.

[0476] The detailed procedure was as follows: 4 μL of 500 mM Hepes-K buffer (pH 7.5), 4 μL of 250 μM Flexizyme (eFx), and 4 μL of 250 μM tRNA analog were added to 12 μL of ultrapure water, heated at 95°C for 3 minutes, and cooled at room temperature for 5 minutes. 8 μL of 3M MgCl2 was added, and the mixture was incubated at room temperature for 5 minutes, followed by incubation on ice for 5 minutes. The acylation reaction of the tRNA analog was initiated by adding 6 μL of DMSO and 2 μL of 100 mM substrate, and then incubated on ice overnight. The reaction was stopped by adding 4 μL of 3M sodium acetate, pH 5.2. This solution was analyzed by 20% denaturing PAGE (50 mM sodium acetate, 6 M urea) under acidic conditions.

[0477] The results of acylation using 2,2,2-trifluoroethyl (S)-2-amino-4-phenylbutanoate (Compound No. 47) and 2,2,2-trifluoroethyl (S)-2-amino-3-(1H-pyrrolo[2,3-b]pyridin-3-yl)propanoate (Compound No. 81) as substrates are shown in Figures 1-3 and 1-4. These results confirmed that amino acid derivatives were efficiently acylated by Flexizyme.

[0478] Test example: Differential scanning calorimetry (DSC) was performed on Compounds 21, 47, 81, and 99 prepared in Example 1-1 to determine the reaction initiation temperature (T DSC ) and reaction heat (Q DSC ) was calculated. The Q values ​​of 2,4-dinitrotoluene (compound number 115) and benzoyl peroxide (compound number 116) were DSC and T DSC The values ​​used were those described in the following literature. DSC Data Collection of Reactive Substances (2) (RIIS-SD-89), https: / / www.jniosh.johas.go.jp / publication / houkoku / houkoku_2007_03_list.html The detailed procedure for differential scanning calorimetry (DSC) is as follows.

[0479] An activated amino acid ester (approximately 2 mg) was sealed in a SUS pressure-resistant cell and set in a heat flux type differential scanning calorimeter. The cell was heated to 500°C at 10°C / min in a nitrogen atmosphere of 50 mL / min. The cell was sealed in air, and an empty cell was used as the reference material. The indium (In) value measured under the same conditions was used as the standard for correction.

[0480] The results are shown in Figure 2. Figure 2 shows the Q values ​​obtained using 2,4-dinitrotoluene (compound number 115) and benzoyl peroxide (compound number 116) as reference compounds. DSC and T DSC The line connecting the two plotted reference points is used as the danger judgment line, and the results show the results of confirming whether or not each activated ester of amino acid is dangerous. In the figure, the dotted line indicates the danger judgment line. If it is on or above the judgment line, it is judged as "dangerous" (class 5 dangerous goods), and if it is below the judgment line, it is judged as "not dangerous" (not class 5 dangerous goods). The horizontal axis is log(T DSC -25), and the vertical axis is logQ DSCIn the figure, -L-Hph-DBE_HCl represents the hydrochloride of 3,5-dinitrobenzyl (S)-2-amino-4-phenylbutanoate (Compound No. 114), Gly-DBE_HCl represents the hydrochloride of 3,5-dinitrobenzyl glycinate (Compound No. 112), -L-Ile-DBE_HCl represents the hydrochloride of 3,5-dinitrobenzyl L-isoleucinate (Compound No. 111), and -L-W7N-DBE_HCl represents the hydrochloride of 3,5-dinitrobenzyl (S)-2-amino-3-(1H-pyrrolo[2,3-b]pyridin-3-yl)propanoate (Compound No. 112). The hydrochloride salt of DBE_HCl (Compound No. 113) is indicated; L-Ile-DCPE_HCl indicates (2,6-dichloropyridin-4-yl)methyl L-isoleucinate (Compound No. 21); L-W7N-TEE_HCl indicates 2,2,2-trifluoroethyl (S)-2-amino-3-(1H-pyrrolo[2,3-b]pyridin-3-yl)propanoate (Compound No. 81); and Gly-DCPE_HCl indicates (2,6-dichloropyridin-4-yl)methyl glycinate (Compound No. 99). L-Hph-DBE_HCl, Gly-DBE_HCl, L-Ile-DBE_HCl, and L-W7N-DBE_HCl were plotted above the risk assessment line, whereas L-Ile-DCPE_HCl, L-W7N-TEE_HCl, and Gly-DCPE_HCl were plotted below the risk assessment line (Figure 2). From the above, it was determined that DCPE and TEE do not fall under the category of dangerous goods of Class 5.

Claims

[Claim 1] The invention as described in the specification or drawings.

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