Inhibitors of BMP‐1/tolloid‐like proteinases: efficacy, selectivity and cellular toxicity

Abstract

BMP ‐1/tolloid‐like proteinases belong to the astacin family of human metalloproteinases, together with meprins and ovastacin. They represent promising targets to treat or prevent a wide range of diseases such as fibrotic disorders or cancer. However, the study of their pathophysiological roles is still impaired by the lack of well‐characterized inhibitors and the questions that remain regarding their selectivity and in vivo efficiency. As a first step towards the identification of suitable tools to be used in functional studies, we have undertaken a systematic comparison of seven molecules known to affect the proteolytic activity of human astacins including three hydroxamates ( FG ‐2575, UK383,367, S33A), the protein sizzled, a new phosphinic inhibitor ( RXP ‐1001) and broad‐spectrum protease inhibitors ( GM 6001, actinonin). Their efficacy in vitro , their cellular toxicity and efficacy in cell cultures were thoroughly characterized. We found that these molecules display very different potency and selectivity profiles, with hydroxamate FG ‐2575 and the protein sizzled being very powerful and selective inhibitors of BMP ‐1, whereas phosphinic peptide RXP ‐1001 behaves as a broad‐spectrum inhibitor of astacins. Their use should therefore be carefully considered in agreement with the aim of the study to avoid result misinterpretation.

BMP-1 disrupts cell adhesion and enhances TGF-β activation through cleavage of the matricellular protein thrombospondin-1

“…This can be avoided by using plates coated with poly-d-lysine (37), but the mechanism underlying this observation has remained unclear. As a first step to elucidate the BMP-1-induced loss of adhesion, we added the synthetic BMP-1 inhibitor S33A [a hydroxamate inhibitor referred to as hxt in this study (31,39)] to the culture medium and observed that the BMP-1-producing cells could be efficiently maintained in the adherent state in the presence of the inhibitor (Fig. 1A).…”

Tolloid‐Like 1 Negatively Regulates Hepatic Differentiation of Human Induced Pluripotent Stem Cells Through Transforming Growth Factor Beta Signaling

“…To clarify the functions of BTPs in hepatic differentiation in the future, it may be helpful to create a TLL1 and BMP1 double‐knockout human iPS cell line. Methods for using BTP‐specific inhibitors may also be effective.…”

Targeting Lysyl Oxidase Family Meditated Matrix Cross-Linking as an Anti-Stromal Therapy in Solid Tumours

“…There has been little research thus far examining the use of BMP1 inhibitors in combination with chemotherapeutics to specifically treat LOX/LOXL1 overexpressing cancers. This may be due, at least in part, to the difficulty in finding inhibitors of BMP1 that are selective, potent and cost effective [ 164 ], and the role that BMP1 plays in a range of other vital cellular processes including bone and cartilage development and repair [ 165 ]. Alternatively, increases in BMP1 may have tumour suppressive effects in Ras transformed cancers, where post-translational processing in cells highly expressing LOX and LOXL1 would increase levels of the tumour suppressive LOX pro-peptide.…”