Methods for the synthesis of arginine-containing peptides

Patent metadata

Field	Value
Publication number	US-11753440-B2
Application number	US-201915734751-A
Country	US
Kind code	B2
Filing date	Jun 5, 2019
Priority date	Jun 5, 2018
Publication date	Sep 12, 2023
Grant date	Sep 12, 2023

Abstract

Methods for the synthesis of arginine-containing peptides are provided. The methods include a deprotection step that minimizes the transfer of by-products deriving from cleaved sulfonyl-5 based side chain protecting groups from arginine to amino acids carrying electron rich side chains.

First claim

The invention claimed is:

1. A method for deprotecting a peptide selected from the group consisting of Boc-Trp-Arg(Pbf)-OH, Ac-Arg(Pbf)-His(Trt)-Phe-OH, TFA-H-Tyr(tBu)-Arg(Pbf)-Pro-OH, Boc-rac-6FTrp-Gly-Arg(Pbf)-Glu(OtBu)-OH, Boc-Tyr(Et)-Arg(Pbf)-Ala-Phe-OH, Ac-5-(OH)Trp-Ala-Arg(Pbf)-Ser(tBu)-Leu-Phe-OH, Boc-Arg(Mtr)-Tyr(tBu)-Phe-OH, Boc-2Nal-Leu-Arg(Pbf)-Phe-OH, Ac-Arg(Pbf)-Met-m(NO 2 )Tyr-Pro-OH and Bz-Gly-His(Trt)-D-Phe-Arg(Pbf)-D-Trp-N(Pr) 2 , wherein the method comprises the steps of:
(a) solubilizing the peptide in a solvent mixture of at least one aprotic organic solvent, a thiol scavenger, and either a protic solvent or a carboxylic acid selected from the group of formic acid, acetic acid and/or trifluoroacetic acid or a mixture thereof, wherein the carboxylic acid is present in an amount of 5 to 30%, based on the total volume of the solvent mixture, followed by
(b) mixing the solution obtained in step (a) with a mixture of (i) a carboxylic acid selected from the group consisting of formic acid, acetic acid and trifluoroacetic acid, and (ii) a sulfonic acid selected from the group consisting of methanesulfonic acid, trifluoromethane sulfonic acid, benzene sulfonic acid and p-toluene sulfonic acid, wherein
a total molar amount of the carboxylic acid used in steps (a) and (b) is higher than a total molar amount of the sulfonic acid.

2. The method according to claim 1 , wherein the at least one aprotic organic solvent in step (a) is selected from the group consisting of aromatic hydrocarbons which are liquid at ambient temperature, C 1-6 -halogenoalkanes and acetonitrile and mixtures thereof.

3. The method according to claim 1 , wherein the aprotic solvent in step (a) is present in an amount of 1 to 100 mL/g peptide.

4. The method according to claim 1 , wherein the protic solvent in step (a) is selected from the group consisting of water, an alcohol and mixtures thereof.

5. The method according to claim 1 , wherein the thiol scavenger is an aliphatic thiol.

6. The method according to claim 1 , wherein step (a) is conducted at a temperature of 15 to 25° C.

7. The method according to claim 1 , wherein the sulfonic acid in step (b) is present in an amount of 1 to 20 mol-equivalents, based on each protected arginine moiety present in the peptide.

8. The method according to claim 1 , wherein a molar ratio of the total amount acid of the carboxylic acid used in steps (a) and (b) to the molar amount of the sulfonic acid used in step (b) is 100:1 to 10:1.

9. The method according to claim 1 , wherein the aprotic solvent in step (a) is present in an amount of 2 to 10 mL/g peptide.

10. The method according to claim 1 , wherein the aprotic solvent in step (a) is present in an amount of 2.5 to 7 mL/g peptide.

11. The method according to claim 1 , wherein the protic solvent in step (a) is selected from the group consisting of water, a C 1-4 -alkyl alcohol and mixtures thereof.

12. The method according to claim 1 , wherein the thiol scavenger is dodecanethiol.

13. The method according to claim 1 , wherein the sulfonic acid in step (b) is present in an amount of 2 to 12 mol-equivalents, based on each protected arginine moiety present in the peptide.

14. The method according to claim 1 , wherein the sulfonic acid in step (b) is present in an amount of 3 to 10 mol-equivalents, based on each protected arginine moiety present in the peptide.

15. The method according to claim 1 , wherein the sulfonic acid in step (b) is present in an amount of 5 to 8 mol-equivalents, based on each protected arginine moiety present in the peptide.

16. The method according to claim 1 , wherein the sulfonic acid used in step (b) is methane sulfonic acid.

17. The method according to claim 8 , wherein the molar ratio of the total amount acid of the carboxylic acid used in steps (a) and (b) to the molar amount of the sulfonic acid used in step (b) is 50:1 to 12:1.

18. The method according to claim 8 , wherein the molar ratio of the total amount acid of the carboxylic acid used in steps (a) and (b) to the molar amount of the sulfonic acid used in step (b) is 30:1 to 15:1.

19. The method according to claim 8 , wherein the molar ratio of the total amount acid of the carboxylic acid used in steps (a) and (b) to the molar amount of the sulfonic acid used in step (b) is 25:1 to 18:1.

Assignees

• Dsm Ip Assets Bv

Inventors

• Heidl Marc
• Geotti-Bianchini Piero

Classifications

• C07K1/064 Primary

  for omega-amino or -guanidino functions · CPC title

• C07K5/1019 Primary

  with the first amino acid being basic · CPC title

• C07K5/06156

  and Trp-amino acid; Derivatives thereof · CPC title

• C07K5/0817

  the first amino acid being Arg · CPC title

• C07K5/1016

  and aromatic or cycloaliphatic · CPC title