METHOD FOR CLEAVAGE OF SOLID PHASE-BOUND PEPTIDES FROM THE SOLID PHASE

• US 20190330266A1
• Filed: 04/10/2019
• Published: 10/31/2019
• Est. Priority Date: 04/10/2018
• Status: Active Grant

• Current Assignee: Sanofi-Aventis Deutschland GmbH (Sanofi )
• Original Assignee: Sanofi-Aventis Deutschland GmbH (Sanofi )
• Inventors: FIEDLER, Wolfgang, PLEUSS, Norbert, HENKEL, Bernd, GERKEN, Manfred
• Granted Patent: US 11,560,402 B2
• Time in Patent Office: Days
• CPC Class Codes: C07K 1/04 on carriers C07K1/003, C07K1/006 take precedence
• CPC Class Codes: C07K 1/042 characterised by the nature of the carrier
• CPC Class Codes: C07K 1/122 Hydrolysis with acids different from HF
• CPC Class Codes: C07K 14/57563 Vasoactive intestinal peptide [VIP]; Related peptides
• CPC Class Codes: C07K 14/605 Glucagons

Abstract

The invention relates to a method for the cleavage of a solid phase-bound polypeptide from the solid phase, the method comprising contacting the solid phase, to which the polypeptide is bound, with a composition consisting essentially of trifluoroacetic acid and 1,2-ethanedithiol, at a temperature in the range of about 23° C. to about 29° C.

Claims

1. A method for the cleavage of a solid phase-bound polypeptide from the solid phase, the method comprising contacting the solid phase, to which the polypeptide is bound, with a composition consisting essentially of trifluoroacetic acid and 1,2-ethanedithiol, at a temperature in the range of about 23°

C. to about 29°

C.

2. The method of claim 1, wherein the solid phase comprises a Rink amide resin.

3. The method of claim 1, wherein the polypeptide is bound to the Rink amide resin by a linker.

4. The method of claim 1, wherein the composition comprises trifluoroacetic acid in an amount of about 95 to about 99% v/v, and 1,2-ethanedithiol in an amount of about 1 to about 5% v/v.

5. The method of claim 1, wherein the composition essentially consists of trifluoroacetic acid in an amount of about 97% v/v, and 1,2-ethanedithiol in an amount of about 3% v/v.

6. The method of claim 1, wherein the composition is contacted with the solid phase to which the polypeptide is bound, at a temperature of about 25°

C. to about 27°

C.

7. The method of claim 1, wherein the composition is contacted with the solid phase to which the polypeptide is bound for 1 to 8 h.

8. The method of claim 1, wherein the polypeptide is selected from GLP-1, analogs and derivatives thereof, exendin-3, analogs and derivatives thereof, and exendin-4, analogs and derivatives thereof.

9. The method of claim 8, wherein the polypeptide is selected from exendin-4, lixisenatide, albiglutide, dulaglutide and semaglutide.

10. A method for the solid-phase synthesis of a polypeptide comprising a pre-determined amino acid sequence, said method comprising:

(a) coupling an amino acid building block, comprising an unprotected C-terminal carboxyl group and a protected N-terminal amino group, C-terminally to a solid phase,

(b) de-protecting the N-terminal amino group of the amino acid building block,

(c) coupling an amino acid building block, comprising an unprotected C-terminal carboxyl group and a protected N-terminal amino group, C-terminally to the unprotected N-terminal amino of step (b),

(d) optionally repeating steps (b) and (c), and

(e) cleaving the polypeptide from the solid phase by the method of claim 1.

11. The method of claim 10, wherein the polypeptide is selected from GLP-1, analogs and derivatives thereof, exendin-3, analogs and derivatives thereof, and exendin-4, analogs and derivatives thereof.

12. The method of claim 10, wherein the polypeptide is selected from exendin-4, lixisenatide, albiglutide, dulaglutide and semaglutide.

16. The method of claim 6, wherein the composition is contacted with the solid phase to which the polypeptide is bound at a temperature of about 26°

C.

17. The method of claim 7, wherein the composition is contacted with the solid phase to which the polypeptide is bound for 3 to 5 h.

18. The method of claim 10, wherein the solid phase is a Rink amide resin.

13. A composition consisting essentially of trifluoroacetic acid and 1,2-ethanedithiol, wherein the composition comprises trifluoroacetic acid in an amount of 95 to 99% v/v, and 1,2-ethanedithiol in an amount of 1 to 5% v/v.

14. The composition of claim 13, essentially consisting of trifluoroacetic acid in an amount of about 97% v/v, and 1,2-ethanedithiol in an amount of about 3% v/v.

15. Use of the composition of claim 13 in a solid-phase synthesis of a polypeptide, wherein the composition is used to cleave the polypeptide from the solid phase.