Method for producing peptide

Patent metadata

Field	Value
Publication number	US-9353148-B2
Application number	US-201314089895-A
Country	US
Kind code	B2
Filing date	Nov 26, 2013
Priority date	May 31, 2011
Publication date	May 31, 2016
Grant date	May 31, 2016

Abstract

The present invention provides a production method of a protected amino acid, protected peptide or peptide, including precipitation and solid-liquid separation of C-protected amino acid or C-protected peptide in a solvent containing water-containing acetonitrile, after removing the N-terminal protecting group from N-protected C-protected amino acid or N-protected C-protected peptide wherein the C-terminal carboxy group is protected by an anchor group.

First claim

The invention claimed is:

1. A method of producing a peptide, comprising:
(1) condensing an N-terminal amino group of (a) a C-protected amino acid or (b) a C-protected peptide, with a C-terminal carboxy group of (c) an N-protected amino acid or (d) an N-protected peptide in a solvent to obtain a reaction solution which comprises an (e) N-protected C-protected peptide,
wherein a C-terminal carboxy group of said (a) C-protected amino acid or said (b) C-protected peptide is protected by an anchor group;
(2) removing the N-terminal protecting group of said (e) N-protected C-protected peptide in said reaction solution, without isolating said N-protected C-protected peptide from said reaction solution, to obtain (b′) a C-protected peptide, wherein when said condensing is conducted with said (b) C-protected peptide, then said (b) C-protected peptide and said (b′) C-protected peptide are different; and
(3) precipitating said C-protected peptide in a solvent comprising a water-acetonitrile mixture, wherein said water-acetonitrile mixture comprises 60 to 95% v/v acetonitrile, based on the total volume of said mixture, after said (2) removing; and
(4) obtaining said C-protected peptide by solid-liquid separation,
wherein said anchor group is a group represented by formula (II):

wherein:
R 1 is a hydrogen atom or, when R b is a group represented by the following formula (a), optionally forms a single bond together with R 3 to form a fluorene ring together with ring A and ring B;
each R 2 is independently an organic group having an aliphatic hydrocarbon group;
p is an integer of 1 to 4;
ring A optionally further has, in addition to OR 2 , a substituent selected from the group consisting of a halogen atom, a C 1-6 alkyl group optionally substituted by a halogen atom, and a C 1-6 alkoxy group optionally substituted by a halogen atom;
R a is a hydrogen atom, or a phenyl group optionally substituted by a halogen atom; and
R b is a hydrogen atom, or a group represented by formula (a):

wherein:
* indicates the position of binding to the remainder of the molecule;
r is an integer of 0 to 4;
each R 4 is independently an organic group having an aliphatic hydrocarbon group;
R 3 is a hydrogen atom, or optionally forms a single bond together with R 1 to form a fluorene ring together with ring A and ring B; and
ring B optionally further has, in addition to OR 4 , a substituent selected from the group consisting of a halogen atom, a C 1-6 alkyl group optionally substituted by a halogen atom, and a C 1-6 alkoxy group optionally substituted by a halogen atom; and
L represent a connection to the carbonyl group of said C-terminus and is —O— or —N(R)—, wherein R is a hydrogen atom, an alkyl group, or an aralkyl group.

2. The method according to claim 1 , further comprising:
(5) removing the C-terminal anchor group of said (b′) C-protected peptide after said (4) obtaining.

3. The method according to claim 1 , wherein, in formula (II), R 1 is a hydrogen atom, R 2 and/or R 4 are/is aliphatic hydrocarbon group(s) having 5 to 60 carbon atoms, p is an integer of 1 to 3, and r is an integer of 0-2.

4. The method according to claim 1 , wherein, in formula (II), R a , R b , and R 1 are each a hydrogen atom, R 2 is an aliphatic hydrocarbon group having 5 to 60 carbon atoms, and p is an integer of 1 to 3.

5. The method according to claim 1 , wherein, in formula (II), R a , R b , and R 1 are each a hydrogen atom, R 2 is an alkyl group having 10 to 40 carbon atoms, and p is 2 or 3.

6. The method according to claim 1 , wherein, in formula (II), R a , R b , and R 1 are each a hydrogen atom, R 2 is an alkyl group having 12 to 30 carbon atoms, and p is 2 or 3.

7. The method according to claim 1 , wherein said anchor group is derived from an anchor compound selected from the group consisting of:
3,4,5-tri(octadecyloxy)benzyl alcohol,
2,4-di(docosyloxy)benzyl alcohol,
4-methoxy-2-[3′,4′,5′-tri(octadecyloxy)benzyloxy]benzyl alcohol,
4-methoxy-2-[3′,4′,5′-tri(octadecyloxy)cyclohexyl-methyloxy]benzyl alcohol,
2-methoxy-4-[3′,4′,5′-tri(octadecyloxy)cyclohexyl-methyloxy]benzyl alcohol,
4-[3′,4′,5′-tri(octadecyloxy)cyclohexylmethyloxy]benzyl alcohol,
3,5-dimethoxy-4-[3′,4′,5′-tri(octadecyloxy)cyclohexyl-methyloxy]benzyl alcohol,
2,4-di(dodecyloxy)benzyl alcohol,
3,4,5-tri(octadecyloxy)benzylamine,
bis(4-docosyloxyphenyl)methanol, and
bis(4-docosyloxyphenyl)methylamine.

8. The method according to claim 1 , wherein said mixture comprises acetonitrile in an amount of 70 v/v% to 90 v/v% based on the the total volume of said mixture.

9. The method according to claim 1 , wherein said mixture comprises acetonitrile in an amount of 75 v/v% to 85 v/v% based on the total volume of said mixture.

10. The method according to claim 1 , wherein said mixture comprises acetonitrile in an amount of 80 v/v% based on the total volume of said mixture.

11. The method according to claim 1 , wherein said solvent comprising a water-acetonitrile mixture is a mixed solvent of said water-acetonitrile mixture, and at least one solvent selected from the group consisting of methanol, ethanol, dimethylformamide, propionitrile, dimethyl sulfoxide, acetone, dichloromethane, chloroform, tetrahydrofuran, cyclopentyl methyl ether, and ethyl acetate.

12. The method according to claim 1 , wherein said solvent is said water-acetonitrile mixture.

13. The method according to claim 1 , wherein said amino-protecting group of said (c) N-protected amino acid or said (d) N-protected peptide is a 9-fluorenylmethyloxycarbonyl group, a tert-butoxycarbonyl group, or a benzyloxycarbonyl group.

14. The method according to claim 1 , wherein the number of amino acid residues of said N-protected peptide is not more than 100.

15. The method according to claim 1 , wherein the number of amino acid residues of said N-protected peptide is not more than 50.

16. The method according to claim 1 , wherein the number of amino acid residues of said N-protected peptide is not more than 30.

17. The method according to claim 1 , wherein the number of amino acid residues of said N-protected peptide is not more than 10.

18. The method according to claim 1 , wherein the number of amino acid residues of said N-protected peptide is not more than 5.

19. The method according to claim 1 , wherein the number of amino acid residues of the C-protected peptide is not more than 100.

20. The method according to claim 1 , wherein the number of amino acid residues of the C-protected peptide is not more than 50.

21. The method according to claim 1 , wherein the number of amino acid residues of the C-protected peptide is not more than 30.

Assignees

• Ajinomoto Kk

Inventors

• Takahashi Daisuke

Classifications

• C07K1/062

  for alpha- or omega-carboxy functions · CPC title

• C07K1/061 Primary

  using protecting groups · CPC title

• Y02P20/55

  Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups · CPC title

• C07C229/34

  the carbon skeleton containing six-membered aromatic rings · CPC title

• C07C229/08 Primary

  the nitrogen atom of the amino group being further bound to hydrogen atoms · CPC title