Non-chromatographic purification of macrocyclic peptides by a resin catch and release

Patent metadata

Field	Value
Publication number	US-11479581-B2
Application number	US-201816618847-A
Country	US
Kind code	B2
Filing date	Jun 8, 2018
Priority date	Jun 9, 2017
Publication date	Oct 25, 2022
Grant date	Oct 25, 2022

Abstract

The disclosure is directed to compounds and methods for preparing purified macrocyclic peptide using “catch-release” methods. These methods comprise reacting a free amino group of a resin-bound linear peptide with an azide- or alkyne-functionalized cap to form a resin-bound capped linear peptide having an azide- or alkyne-functionalized cap; cleaving the capped linear peptide from the resin to form a free capped linear peptide having an azide- or alkyne-functionalized cap; reacting the free capped linear peptide having an azide-functionalized cap with an alkyne-functionalized catch resin, or reacting the free capped linear peptide having an akynyl-functionalized cap with an azide functionalized catch resin, to form a catch-resin bound capped linear peptide; reacting the catch-resin bound capped linear peptide under conditions sufficient to effect macrocyclization of the linear peptide and release of the macrocyclic peptide from the catch resin.

First claim

What is claimed:

1. A catch-release method of preparing a purified macrocyclic peptide comprising
(a) preparing a resin-bound linear peptide wherein said linear peptide comprises a free amino group and an amino acid residue having a nucleophilic side chain;
(b) reacting the free amino group of the resin-bound linear peptide with an azide- or alkyne-functionalized cap to form a resin-bound capped linear peptide having an azide- or alkyne-functionalized cap;
(c) cleaving said capped linear peptide from the resin to form a free capped linear peptide having an azide- or alkyne-functionalized cap;
(d) reacting the free capped linear peptide having an azide-functionalized cap with an alkyne functionalized catch resin, or reacting the free capped linear peptide having an alkyne-functionalized cap with an azide functionalized catch resin, to form a catch-resin bound capped linear peptide;
(e) washing the catch-resin bound capped linear peptide;
(f) reacting the catch-resin bound capped linear peptide under conditions sufficient to effect macrocyclization of the linear peptide and release of the macrocyclic peptide from the catch resin.

2. The method of claim 1 , wherein the macrocyclization and release results from reaction of the nucleophilic side chain of the amino acid in the linear peptide with an electrophilic leaving group moiety on the cap.

3. The method of claim 1 , wherein the azide- or alkyne-functionalized cap is

4. The method of claim 1 , wherein the catch resin is

5. The method of claim 1 , wherein step (d) is performed in the presence of 1,4-dithiothreitol.

6. The method of claim 1 , further comprising the step of reacting the catch-resin bound capped linear peptide with an azide prior to subjecting the catch-resin bound capped linear peptide to the conditions that effect macrocyclization and release.

7. The method of claim 1 , wherein the conditions that effect the macrocyclization of the linear peptide and release of the macrocyclic peptide from the catch resin in step (f) are treatment with 0.1 M NH 4 OAc-0.1 M NH 3 in methanol.

Assignees

• Bristol Myers Squibb Co

Inventors

• Creech Gardner S
• Kheirabadi Mahboubeh
• Eastgate Martin D
• Nirschl David S
• Carter Percy H

Classifications

• C07K1/1077

  by covalent attachment of residues other than amino acids or peptide residues, e.g. sugars, polyols, fatty acids · CPC title

• C07K1/042

  characterised by the nature of the carrier · CPC title

• C07C309/77 Primary

  containing carboxyl groups bound to the carbon skeleton · CPC title

• C07C309/75

  containing singly-bound oxygen atoms bound to the carbon skeleton · CPC title

• C07C317/18

  with sulfone or sulfoxide groups bound to acyclic carbon atoms of the carbon skeleton · CPC title