	US 12,351,601 B2
	Method for producing peptide continuously
Daisuke Takahashi, Kawasaki (JP); Metten Bert, Wettern (BE); Kimio Kawajiri, Kawasaki (JP); and Ryotaro Nakaya, Wetteren (BE)
Assigned to AJINOMOTO CO., INC., Tokyo (JP)
Filed by AJINOMOTO CO., INC., Tokyo (JP)
Filed on Oct. 21, 2021, as Appl. No. 17/506,934.
Application 17/506,934 is a continuation of application No. PCT/JP2020/017670, filed on Apr. 24, 2020.
Claims priority of application No. 2019-084652 (JP), filed on Apr. 25, 2019.
Prior Publication US 2022/0041648 A1, Feb. 10, 2022
Int. Cl. C07K 1/06 (2006.01)

CPC C07K 1/062 (2013.01)

15 Claims

1. A method for producing a peptide by flow synthesis in a flow reactor wherein the peptide is protected by a pseudo-solid phase protecting group to maintain solubility of the peptide as it elongates, comprising the following (A) or (B), or a combination thereof:

(A) producing a reaction mix comprising the peptide protected by a pseudo-solid-phase group in a flow reactor; wherein the peptide comprises an N-protected C-protected peptide having an amino acid residue number of from 5 to 100, in which an N-terminal amino group and C-terminal are protected by protecting groups;

washing the reaction mix that contains the peptide with water and/or a hydrophilic organic solvent

and then

separating the peptide into an organic layer via an oil and water type phase separation comprising water or the hydrophilic organic solvent and a solvent immiscible with water or the hydrophilic organic solvent, thereby recovering the protected peptide,

wherein protected peptide solution is diluted 5- to 100-fold compared to the initial concentration of the substrate peptide in the immiscible solvent

(B) producing a reaction mix comprising the peptide protected by a pseudo-solid-phase group in a flow reactor; wherein the peptide comprises an N-terminal amino group that is not protected, a C-terminal that is protected by a protecting group having an amino acid residue number of from 5 to 100, in which an N-terminal amino group is not protected and C-terminal are protected by protecting groups:

washing the reaction mix that contains the peptide with water and/or a hydrophilic organic solvent; separating the peptide into an organic layer via an oil and water type phase separation comprising water or the hydrophilic organic solvent and a second solvent immiscible with water or immiscible with the hydrophilic organic solvent, thereby recovering the protected peptide,

wherein protected peptide solution is diluted 2- to 100-fold compared to the initial concentration of the substrate peptide in the immiscible solvent