Method for removing FMOC group

Patent metadata

Field	Value
Publication number	US-9334302-B2
Application number	US-201414305841-A
Country	US
Kind code	B2
Filing date	Jun 16, 2014
Priority date	Dec 15, 2011
Publication date	May 10, 2016
Grant date	May 10, 2016

Abstract

The present invention relates to a method of removing an Fmoc group, including a step of mixing a compound represented by the formula (I):

HS-L-COOH  (I)

wherein L is a C 1-8 alkylene group optionally having substituent(s), an amino group-containing compound protected by an Fmoc group, and a base to give a reaction mixture containing a compound represented by the formula (II):

Fm—S-L-COOH  (II)

wherein Fm is a 9-fluorenylmethyl group, and L is as mentioned above, and an amino group-containing compound, and a step of removing the compound represented by the formula (II) by washing the obtained reaction mixture with a basic aqueous solution. According to the present invention, a removal method of Fmoc group, which can remove a dibenzofulvene derivative as a byproduct with ease, can be provided.

First claim

The invention claimed is:

1. A method of removing an Fmoc group, comprising:
mixing an amino group-containing compound protected by an Fmoc group, a base, and a compound represented by formula (I):
HS-L-COOH  (I)

where L is a C 1-8 alkylene group optionally having at least one substituent, to obtain a reaction mixture comprising an amino group-containing compound and a compound represented by formula (II):
Fm—S-L-COOH  (II)

where Fm is a 9-fluorenylmethyl group; and
washing the reaction mixture with a basic aqueous solution such that the compound represented by the formula (II) is removed;
wherein the amino group-containing compound protected by an Fmoc group, and the amino group-containing compound, do not have a free carboxy group.

2. The method according to claim 1 , wherein the base is an organic base.

3. The method according to claim 2 , wherein the organic base is 1,8-diazabicyclo[5.4.0]-7-undecene.

4. The method according to claim 1 , wherein the compound represented by the formula (I) is at least one selected from the group consisting of 3-mercaptopropionic acid, thiomalic acid and cysteine.

5. The method according to claim 1 , wherein the basic aqueous solution is an aqueous solution of at least one selected from the group consisting of lithium carbonate, potassium carbonate, sodium carbonate, lithium hydrogen carbonate, potassium hydrogen carbonate, sodium hydrogen carbonate, lithium hydroxide, potassium hydroxide and sodium hydroxide.

6. The method according to claim 1 , wherein the amino group-containing compound protected by an Fmoc group is N-Fmoc-C-protected peptide, N-Fmoc-C-protected amino acid or N-Fmoc-C-protected amino acid amide, and
the amino group-containing compound in the reaction mixture is C-protected peptide, C-protected amino acid or C-protected amino acid amide.

7. A method of producing a peptide by a liquid phase synthesis method, comprising:
removing an Fmoc group by the method according to claim 6 .

8. The method according to claim 7 , further comprising:
condensing C-protected peptide, C-protected amino acid or C-protected amino acid amide, and N-Fmoc amino acid or N-Fmoc peptide in the presence of a condensing agent to obtain the N-Fmoc-C-protected peptide.

9. The method according to claim 8 , wherein the condensing is performed in the presence of an activator and the condensing agent.

10. The method according to claim 8 , wherein the C-protected peptide, C-protected amino acid or C-protected amino acid amide is obtained by removing the Fmoc group, and subjected to the condensing without isolating as a solid.

11. The method according to claim 10 , wherein the peptide is produced by one-pot synthesis.

12. The method according to claim 7 , further comprising:
condensing C-protected peptide, C-protected amino acid or C-protected amino acid amide, and N-Fmoc amino acid active ester or N-Fmoc peptide active ester to obtain the N-Fmoc-C-protected peptide.

13. The method according to claim 12 , wherein the C-protected peptide, C-protected amino acid or C-protected amino acid amide is obtained by removing the Fmoc group, and subjected to the condensing without isolating as a solid.

14. The method according to claim 13 , wherein the peptide is produced by one-pot synthesis.

15. The method according to claim 7 , further comprising:
(1) condensing C-protected peptide, C-protected amino acid or C-protected amino acid amide, and N-Fmoc amino acid or N-Fmoc peptide in the presence of a condensing agent; and
(2) condensing C-protected peptide, C-protected amino acid or C-protected amino acid amide, and N-Fmoc amino acid active ester or N-Fmoc peptide active ester to obtain the N-Fmoc-C-protected peptide.

16. The method according to claim 15 , wherein the (1) condensing is performed in the presence of an activator and the condensing agent.

17. The method according to claim 15 , wherein the C-protected peptide, C-protected amino acid or C-protected amino acid amide is obtained by removing the Fmoc group, and subjected to the (1) condensing and the (2) condensing without isolating as a solid.

18. The method according to claim 17 , wherein the peptide is produced by one-pot synthesis.

Assignees

• Ajinomoto Kk

Inventors

• Takahashi Daisuke

Classifications

• C07K1/063 Primary

  for alpha-amino functions · CPC title

• C07K1/064

  for omega-amino or -guanidino functions · CPC title

• C07K1/08

  using activating agents {(C07K1/003, C07K1/006 take precedence)} · CPC title

• C07C227/04

  Formation of amino groups in compounds containing carboxyl groups · CPC title

• C07B43/04

  of amino groups · CPC title