# US 2023/0056969 A1 · Method for synthesizing peptide compound

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- Number: US 2023/0056969 A1
- Document ID: US20230056969A1
- Status: Published application
- Kind: A1
- Application number: 17/788,506
- Filed: 2020-12-25
- Published: 2023-02-23
- Earliest claimed: 2019-12-27
- Applicants: Chugai Seiyaku Kabushiki Kaisha
- Inventors: Yasuhiro KONDO; Kentarou SETO; Shio KOMIYA; Zengye HOU; Masatoshi MURAKATA
- Web page: https://patentgrep.com/patent/US20230056969A1

## CPC

- C07K 1/06: using protecting groups or activating agents {(C07K1/003, C07K1/006 take precedence)}
- C07K 1/02: in solution {(C07K1/003, C07K1/006 take precedence)}
- Y02P 20/55: Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- Y02P: CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS

## Abstract

The present inventors found that, in the synthesis of a peptide compound involving condensation of a C-terminal-activated substance of an acid component with an amine component, the C-terminal-activated substance can be removed by mixing a solution containing a residual C-terminal-activated substance after a condensation reaction with a tertiary amine and water or an aqueous solution.

## Claims

### Claim 1

A method of producing a peptide compound, comprising:
step A: a step of obtaining a reaction mixture comprising a peptide compound obtained by condensing a C-terminal-activated substance of an acid component with an amine component in a solvent; and
step B: a step of mixing the reaction mixture, a tertiary amine, and water or an aqueous solution to remove the C-terminal-activated substance.

### Claim 2 (depends on 1)

The method of claim 1, wherein the tertiary amine is nucleophilic to the C-terminal-activated substance.

### Claim 3 (depends on 1)

The method of claim 1, wherein the tertiary amine is an amine having small steric hindrance in the vicinity of nitrogen.

### Claim 4 (depends on 1)

The method of claim 1, wherein the tertiary amine is NMI, DMAP, or trimethylamine.

### Claim 5 (depends on 1)

The method of claim 1, wherein the peptide compound comprises one or more non-natural amino acids.

### Claim 6 (depends on 1)

The method of claim 1, wherein a temperature for allowing the tertiary amine to act on the C-terminal-activated substance is 25° C. to 60° C.

### Claim 7 (depends on 1)

The method of claim 1, wherein the tertiary amine is added in an amount of 0.5 equivalents or more relative to the amine component.

### Claim 8 (depends on 1)

The method of claim 1, wherein the C-terminal-activated substance is allowed to be acted on by the tertiary amine and hydrolyzed, and is removed.

### Claim 9 (depends on 1)

The method of claim 1, wherein step B further comprises separating the reaction mixture into an organic layer and an aqueous layer and then washing the organic layer, and wherein a residual amount of the C-terminal-activated substance after the washing is 1.0% or less.

### Claim 10 (depends on 1)

The method of claim 1, wherein the solvent in step A is toluene, acetonitrile, tetrahydrofuran, 2-methyltetrahydrofuran, isopropyl acetate, ethyl acetate, methyl tert-butyl ether, cyclopentyl methyl ether, or N,N-dimethylformamide, or a mixed solvent thereof.

### Claim 11 (depends on 1)

The method of claim 1, wherein in step B, the aqueous solution is an aqueous potassium carbonate solution or an aqueous sodium carbonate solution.

### Claim 12 (depends on 1)

The method of claim 1, wherein the acid component is a first amino acid having an amino group protected with a protecting group, and wherein a side chain of the first amino acid comprises one or more carbon atoms.

### Claim 13 (depends on 12)

The method of claim 12, wherein the side chain is optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted alkoxyalkyl, optionally substituted cycloalkylalkyl, optionally substituted aralkyl, or optionally substituted heteroarylalkyl.

### Claim 14 (depends on 1)

The method of claim 1, wherein a time for allowing the tertiary amine to act on the C-terminal-activated substance is 2 hours or less.

### Claim 15 (depends on 1)

The method of claim 1, wherein the C-terminal-activated substance is formed in the presence of a condensing agent, and wherein the condensing agent comprises T3P, HATU, BEP, DMT-MM, a combination of EDC and PfpOH, a combination of EDC and HOOBt, or a combination of EDC and HOBt.