Phosphono Bisbenzguanidines as Irreversible Dipeptidomimetic Inhibitors and Activity-Based Probes of Matriptase-2.

Abstract

Matriptase-2, a type II transmembrane serine protease, plays a key role in human iron homeostasis. Inhibition of matriptase-2 is considered as an attractive strategy for the treatment of iron-overload diseases, such as hemochromatosis and β-thalassemia. In the present study, synthetic routes to nine dipeptidomimetic inactivators were developed. Five active compounds (41-45) were identified and characterized kinetically as irreversible inhibitors of matriptase-2. In addition to a phosphonate warhead, these dipeptides possess two benzguanidine moieties as arginine mimetics to provide affinity for matriptase-2 by binding to the S1 and S3/S4 subpockets, respectively. This binding mode was strongly supported by covalent docking analysis. Compounds 41-45 were obtained as mixtures of two diastereomers and were therefore separated into the single epimers. Compound 45 A, with S configuration at the N-terminal amino acid and R configuration at the phosphonate carbon atom, was the most potent matriptase-2 inactivator with a rate constant of inactivation of 2790 m(-1)  s(-1) and abolished the activity of membrane-bound matriptase-2 on the surface of intact cells. Based on the chemotyp of phosphono bisbenzguanidines, the design and synthesis of a fluorescent probe (51 A) by insertion of a coumarin label is described. The in-gel fluorescence detection of matriptase-2 was demonstrated by applying 51 A as the first activity-based probe for this enzyme.

• Authors: Daniela Häußler, Martin Mangold, Norbert Furtmann, Annett Braune, Michael Blaut, Jürgen Bajorath, Marit Stirnberg, Michael Gütschow
• Journal: Chemistry (Weinheim an der Bergstrasse, Germany) (Chemistry) Vol. 22 Issue 25 Pg. 8525-35 (06 13 2016) ISSN: 1521-3765 [Electronic] Germany
• PMID: 27214780 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Chemical References

• Coumarins
• Fluorescent Dyes
• Guanidines
• Membrane Proteins
• Peptidomimetics
• Serine Proteinase Inhibitors
• Phosphorus
• coumarin
• Serine Endopeptidases
• matriptase 2
• Trypsin
• Factor Xa

Topics

• Animals
• Binding Sites
• Catalytic Domain
• Cattle
• Coumarins (chemistry)
• Factor Xa (chemistry, metabolism)
• Fluorescent Dyes (chemistry)
• Guanidines (chemical synthesis, chemistry, metabolism)
• Humans
• Kinetics
• Membrane Proteins (antagonists & inhibitors, metabolism)
• Molecular Docking Simulation
• Peptidomimetics
• Phosphorus (chemistry)
• Serine Endopeptidases (metabolism)
• Serine Proteinase Inhibitors (chemical synthesis, chemistry, metabolism)
• Stereoisomerism
• Trypsin (chemistry, metabolism)