Cleavable linker for peptide synthesis

Patent metadata

Field	Value
Publication number	US-11548911-B2
Application number	US-202117170060-A
Country	US
Kind code	B2
Filing date	Feb 8, 2021
Priority date	Aug 9, 2018
Publication date	Jan 10, 2023
Grant date	Jan 10, 2023

Abstract

The present invention provides a new building block for peptide synthesis, which introduces a cleavage site that can be used to generate cleavable fragments subsequent to a peptide sequence.

First claim

The invention claimed is:

1. A method comprising:
a) synthesizing a peptide on a first solid support, said peptide comprising a first terminal amino group,
b) providing a building block comprising the structure:

wherein 2≤n≤24, m=2 or 3,
A is an acid labile protective group, and
B is a tag or base labile protective group; and
c) coupling said building block to said first terminal amino group of said peptide.

2. The method of claim 1 , wherein when B is a base labile protective group further comprising:
c) removing the base labile protective group B from the building block to provide a deprotected amino group, and
d) coupling at least one amino acid building block to the deprotected amino group of the building block, wherein the at least one amino acid building block comprises a second terminal amino group.

3. The method of claim 1 , wherein when B is a base labile protective group further comprising:
c) removing the base labile protective group B from the building block to provide a deprotected amino group,
e) optionally coupling at least one amino acid building block to the deprotected amino group of the building block, wherein the at least one amino acid building block comprises a second terminal amino group, and
e) coupling a tag or a functional group to either (i) the second terminal amino group of the at least one amino acid building block, or (ii) the deprotected amino group of the building block.

4. The method of claim 3 , wherein said tag or functional group is selected from the group consisting of PEG, poly-lysine, poly-arginine, poly-glutamic acid, poly-aspartic acid, biotin, hydrazine, aminooxy, azide, alkynyl, alkenyl, aldehyde, pyrroloalanine, carboxy, and thiol.

5. The method of claim 1 , further comprising:
removing the acid labile protective group A from the building block at a pH≤6, thereby also removing other protective groups present on said peptide and cleaving said peptide from the first solid support.

6. The method of claim 5 , further comprising:
contacting said cleaved peptide with a buffer having a pH≥8.

7. The method of claim 3 , wherein said tag or functional group is a solubilizing tag, and wherein the method further comprises:
g) removing the acid labile protective group A from the building block at a pH≤6, thereby also removing other protective groups present on said peptide and cleaving said peptide from the first solid support,
h) purifying said cleaved peptide, and
i) cleaving said solubilizing tag at a pH≥8.

8. The method of claim 3 , wherein said tag or functional group is an immobilizing tag, and wherein said method further comprises:
g) removing the acid labile protective group A from the building block at a pH≤6, thereby also removing other protective groups present on said peptide and cleaving said peptide from the first solid support,
h) immobilizing said cleaved peptide via said immobilizing tag on a second solid support,
i) optionally conjugating said cleaved peptide to an additional chemical entity, and
j) cleaving said immobilizing tag at a pH≥8.

9. The method of claim 8 , wherein said additional chemical entity is selected from the group consisting of a nucleic acid, an oligonucleotide, and a nucleotide.

10. The method of claim 9 , wherein said nucleotide is a nucleoside-hexaphosphate.

Assignees

• Roche Sequencing Solutions Inc

Inventors

• Bergmann Frank
• Loibl Simon Ferdinand
• Pomplun Sebastian Johannes

Classifications

• C07C271/22 Primary

  to carbon atoms of hydrocarbon radicals substituted by carboxyl groups · CPC title

• C07K1/04 Primary

  on carriers {(C07K1/003, C07K1/006 take precedence)} · CPC title

• C07K14/001

  by chemical synthesis · CPC title

• Y02P20/55

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