US 10,883,132 B2Granted

Chemo-enzymatic synthesis of Semaglutide, Liraglutide and GLP-1

AssigneeEnzypep B.V.

InventorsTimo Nuijens, Ana Toplak, Peter Jan Leonard Mario Quaedflieg

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Abstract

A method for synthesizing a peptide having the sequence His-X-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Y-Glu-Phe-Ile-Ala-Trp-Leu-Val-Z-Gly-Arg-Gly is disclosed. The method includes enzymatically coupling: (a) a peptide C-terminal ester or thioester having a first peptide fragment with the sequence His-X-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-(thio)ester; and (b) a peptide nucleophile having an N-terminally unprotected amine having a second peptide fragment with the sequence H-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Y-Glu-Phe-Ile-Ala-Trp-Leu-Val-Z-Gly-Arg-Gly; wherein: X is Ala or an α-amino-isobutyric acid (Aib) residue; Y is Lys, which Lys has a free side-chain ε-amino group or of which Lys the side-chain ε-amino group is protected with a protective group or of which Lys the side-chain ε-amino group is functionalized with an amino acid or another functional group; and Z is Arg or Lys.

Claims20

Dependent claims

A method for synthesizing a peptide comprising the sequence His-X-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Y-Glu-Phe-Ile-Ala-Trp-Leu-Val-Z-Gly-Arg-Gly, comprising enzymatically coupling

(a) a peptide C-terminal ester or thioester comprising a first peptide fragment comprising the sequence His-X-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-(thio)ester (SEQ ID NO:
11);
and
(b) a peptide nucleophile having an N-terminally unprotected amine comprising a second peptide fragment comprising the sequence H-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Y-Glu-Phe-Ile-Ala-Trp-Leu-Val-Z-Gly-Arg-Gly (SEQ ID NO:
12)
wherein
X is Ala or an a-amino-isobutyric acid (Aib) residue;
Y is Lys, which Lys has a free side-chain s-amino group or of which Lys the side-chain s-amino group is protected with a protective group or of which Lys the side-chain s-amino group is functionalized with an amino acid or another functional group, in particular a functional group selected from the group consisting of γ-Glu-OH, Pal-γ-Glu-OH, AEEA-AEEA-γ-Glu-OH and AEEA-AEEA-γγ-Glu-N-17-carboxyheptadecanoyl-OH, wherein Pal is palmitoyl and AEEA-AEEA is -2-[2-(2-aminoethoxy)ethoxy]acetyl-2-[2-(2-aminoethoxy)ethoxy]acetyl;
and
Z is Arg or Lys;
which enzymatic coupling is catalyzed by a ligase.

8 more elements

Dependencies are inferred from the claim text.