ABSTRACT
BACKGROUND: The metalloproteinases ADAM10 and ADAM17 are involved in various diseases: neurodegeneration, cancer and inflammation. OBJECTIVE: The inhibition of these proteases is a promising target in the treatment of inflammation and cancer. METHODS AND RESULTS: In this study, we present an improved synthesis of the ADAM10 reference inhibitor GI254023X with a higher overall yield, enhanced detection ability and increased acid stability, providing easier handling. CONCLUSION: This upscaled synthesis, free of diastereomeric intermediates, ensures single-batch identity, thus warranting its reproducibility in further biological investigations.
Subject(s)
ADAM Proteins/antagonists & inhibitors , Amyloid Precursor Protein Secretases/antagonists & inhibitors , Dipeptides/chemical synthesis , Dipeptides/pharmacology , Hydroxamic Acids/chemical synthesis , Hydroxamic Acids/pharmacology , Inflammation Mediators/chemical synthesis , Membrane Proteins/antagonists & inhibitors , Protease Inhibitors/chemical synthesis , ADAM Proteins/metabolism , ADAM10 Protein , Amides/chemical synthesis , Amides/chemistry , Amyloid Precursor Protein Secretases/metabolism , Antineoplastic Agents/chemical synthesis , Antineoplastic Agents/pharmacology , Cell Line , Humans , Inflammation/drug therapy , Inflammation/enzymology , Inflammation Mediators/pharmacology , Membrane Proteins/metabolism , Neoplasms/drug therapy , Neoplasms/enzymology , Neoplasms/pathology , Protease Inhibitors/pharmacology , Reproducibility of ResultsABSTRACT
Syndecans are cell surface proteoglycans that bind and modulate various proinflammatory mediators and can be proteolytically shed from the cell surface. Within the lung, syndecan-1 and -4 are expressed as transmembrane proteins on epithelial cells and released in the bronchoalveolar fluid during inflammation. We here characterize the mechanism leading to the generation of soluble syndecan-1 and -4 in cultured epithelial cells and murine lung tissue. We show that the bladder carcinoma epithelial cell line ECV304, the lung epithelial cell line A459 and primary alveolar epithelial cells express and constitutively release syndecan-1 and -4. This release involves the activity of the disintegrin-like metalloproteinase ADAM17 as demonstrated by use of specific inhibitors and lentivirally transduced shRNA. Stimulation of epithelial cells with PMA, thrombin, or proinflammatory cytokines (TNFalpha/IFNgamma) led to the down-regulation of surface-expressed syndecan-1 and -4, which was associated with a significant increase of soluble syndecans and cell-associated cleavage fragments. The enhanced syndecan release was not related to gene induction of syndecans or ADAM17, but rather due to increased ADAM17 activity. Soluble syndecan-1 and -4 were also released into the bronchoalveolar fluid of mice. Treatment with TNFalpha/IFNgamma increased ADAM17 activity and syndecan release in murine lungs. Both constitutive and induced syndecan shedding was prevented by the ADAM17 inhibitor. ADAM17 may therefore be an important regulator of syndecan functions on inflamed lung epithelium.
Subject(s)
ADAM Proteins/metabolism , Epithelial Cells/enzymology , Inflammation/enzymology , Lung/cytology , Syndecan-1/metabolism , Syndecan-4/metabolism , ADAM17 Protein , Animals , Cell Line, Tumor , Humans , In Vitro Techniques , Mice , Mice, Inbred C57BL , Peptide Fragments/metabolismABSTRACT
2-Alkenylchroman-4-ones, 2-alkenylthiochroman-4-ones, and 2-alkenylquinol-4-ones were prepared with very good regioselectivity by Me3SiOTf-mediated conjugate addition of alkenylmagnesium bromides and alkenyllithium compounds to chromones thiochromones, and quinol-4-ones. A number of products exhibit a considerable antimicrobial activity. The best activity, with respect to the spectrum of antimicrobial activity, was observed for 2-vinylchroman-4-ones containing an unsubstituted vinyl group and a chloride group located at the chromanone moiety.
