Details

Autor(en) / Beteiligte

• Kasper, Marc‐André
• Gerlach, Marcus
• Schneider, Anselm F. L.
• Groneberg, Christiane
• Ochtrop, Philipp
• Boldt, Stefanie
• Schumacher, Dominik
• Helma, Jonas
• Leonhardt, Heinrich
• Christmann, Mathias
• Hackenberger, Christian P. R.

Titel

N‐Hydroxysuccinimide‐Modified Ethynylphosphonamidates Enable the Synthesis of Configurationally Defined Protein Conjugates

Ist Teil von

• Chembiochem : a European journal of chemical biology, 2020-01, Vol.21 (1-2), p.113-119

Ort / Verlag

Germany: Wiley Subscription Services, Inc

Erscheinungsjahr

2020

Link zum Volltext

Quelle

Wiley Online Library - AutoHoldings Journals

Beschreibungen/Notizen

• Herein, the application of N‐hydroxysuccinimide‐modified phosphonamidate building blocks for the incorporation of cysteine‐selective ethynylphosphonamidates into lysine residues of proteins, followed by thiol addition with small molecules and proteins, is reported. It is demonstrated that the building blocks significantly lower undesired homo‐crosslinking side products that can occur with commonly applied succinimidyl 4‐(N‐maleimidomethyl)cyclohexane‐1‐carboxylate (SMCC) under physiological pH. The previously demonstrated stability of the phosphonamidate moiety additionally solves the problem of premature maleimide hydrolysis, which can hamper the efficiency of subsequent thiol addition. Furthermore, a method to separate the phosphonamidate enantiomers to be able to synthesize protein conjugates in a defined configuration has been developed. Finally, the building blocks are applied to the construction of functional antibody–drug conjugates, analogously to FDA‐approved, SMCC‐linked Kadcyla, and to the synthesis of a functional antibody–protein conjugate. Creating links: Building blocks equipped with an N‐hydroxysuccinimide and an ethynylphosphonamidate residue can be used as selective amine–thiol linkers with defined configuration. The high selectivity for cysteine can solve current issues of succinimidyl 4‐(N‐maleimidomethyl)cyclohexane‐1‐carboxylate associated with homo‐crosslinking side products under physiological conditions.