Pyridine-Containing Macrocycles Display MMP-2/9 Inhibitory Activity and Distinct Effects on Migration and Invasion of 2D and 3D Breast Cancer Models.

Susana Proença,Joana P Miranda,Filipa Ramilo-Gomes,Rita C Guedes,Nuno G Oliveira,M Fátima Cabral,Bernardo Antunes,Ana S Fernandes,Matilde Castro,Judite Costa

doi:10.3390/ijms20205109

Abstract

The role of metalloproteinases (MMPs) on the migration and invasion of cancer cells has been correlated with tumor aggressiveness, namely with the up-regulation of MMP-2 and 9. Herein, two pyridine-containing macrocyclic compounds, [15]pyN5 and [16]pyN5, were synthesized, chemically characterized and evaluated as potential MMP inhibitors for breast cancer therapy using 3D and 2D cellular models. [15]pyN5 and [16]pyN5 (5–20 µM) showed a marked inhibition of MMPs activity (100% at concentrations ≥ 7.5 μM) when compared to ARP-100, a known MMP inhibitor. The inhibitory activity of [15]pyN5 and [16]pyN5 was further supported through in silico docking studies using Goldscore and ChemPLP scoring functions. Moreover, although no significant differences were observed in the invasion studies in the presence of all MMPs inhibitors, cell migration was significantly inhibited by both pyridine-containing macrocycles at concentrations above 5 μM in 2D cells (p < 0.05). In spheroids, the same effect was observed, but only with [16]pyN5 at 20 μM and ARP-100 at 40 μM. Overall, [15]pyN5 and [16]pyN5 led to impaired breast cancer cell migration and revealed to be potential inhibitors of MMPs 2 and 9.

Highlights

Matrix metalloproteinases (MMPs) have been found to correlate with increased breast tumor aggressiveness [1,2,3,4,5]
Viable Spheroids were Obtained from MDA-MB-231 Cells
Spheroid diameter increased during culture time, reaching a diameter of 191 μm at day 4 of culture (Figure 3A,B)

Summary

Introduction

Matrix metalloproteinases (MMPs) have been found to correlate with increased breast tumor aggressiveness [1,2,3,4,5]. Tumor cells recruit ECM (extracellular matrix) proteases, such as MMPs, to their leading edge for localized proteolysis of the ECM net [1,2,3,4,5]. MMPs are highly conserved proteins containing a structural Zn2+ ion and one to three Ca2+ ions, and in their catalytic domain a catalytic Zn2+ ion coordinated through the conserved motif HExGHxxGxxH [6]. Other common structures to MMPs are their hemopexin-like C-terminal domain essential for its non-proteolytic functions, and a pro-peptide domain which coordinates the catalytic Zn2+ ion maintaining MMPs latency, being its cleavage required for activation [6,7].

Methods

Results

Conclusion