3-Amidinophenylalanine-derived matriptase inhibitors can modulate hepcidin production in vitro

Erzsébet Pászti-Gere,Gergely Szombath,András Székács,Michael Gütschow,Torsten Steinmetzer

doi:10.1007/s00210-019-01743-x

Abstract

Matriptase-2 (MT-2) is a type II transmembrane serine protease and predominantly attached to the surface of hepatocytes. MT-2 decreases the production of hepcidin, a key regulator of iron homeostasis. In this study, the effects of four 3-amidinophenylalanine-derived combined matriptase-1/matriptase-2 (MT-1/2) inhibitors (MI-432, MI-441, MI-460, and MI-461) on hepcidin production were investigated in hepatocyte mono- and hepatocyte-Kupffer cell co-cultures. In MI-461-treated cell cultures, the extracellular hydrogen peroxide contents and the interleukin-6 and -8 (IL-6 and IL-8) levels were determined and compared to controls. Hepcidin overproduction was observed in hepatocytes upon treatment with MI-432, MI-441 and MI-461 at 50 μM. In contrast, extracellular hydrogen peroxide levels were not elevated significantly after matriptase inhibition with MI-461. Furthermore, MI-461 did not induce increases in IL-6 and IL-8 levels in these hepatic models. A model of the binding mode of inhibitor MI-461 in complex with MT-2 revealed numerous polar contacts contributing to the nanomolar potency of this compound. Based on the in vitro data on hepcidin regulation, treatment with MI-461 might be valuable in pathological states of iron metabolism without causing excessive oxidative stress.

Highlights

Hepcidin is mainly produced in the liver and acts as a key regulator of iron homeostasis
Hepatocytes in mono-cultures (Hep) mono- and HepK6 co-cultures embedded in the presence or absence of hydrogel matrices were exposed to 50 μM of the dual MT-1/2 inhibitors (MI-432, MI-441, MI-460, and MI461) for 24 h, and the data were compared to untreated samples (Fig. 3)
No significant changes were observed in hepcidin levels in 2D and hydrogel-assisted hepatocyte-Kupffer cell co-cultures exposed to the MT-1/2 inhibitors (p > 0.05) (Fig. 3c and d)

Summary

Introduction

Hepcidin is mainly produced in the liver and acts as a key regulator of iron homeostasis. It prevents excessive iron from entering the blood circulation via inhibiting the absorption of nutritional iron from enterocytes and suppressing the release of iron. When the blood iron concentration is high, hepcidin production in the liver becomes elevated and contrarily, in the case of the iron shortage, the hormone secretion decreases significantly leading to an enhanced iron transport (Ganz and Nemeth 2012). Germany 4 Agro-Environmental Research Institute, National Agricultural. Hepcidin prevents iron overload (Knutson et al 2005) which could lead to the formation of reactive oxygen species (ROS) or to severe damages to subcellular components such as electron transport chain of respiration or mitochondrial DNA content. Hepcidin synthesis in the liver is upregulated by increasing levels of plasma iron supported by the fact that iron-transferrin saturation can stimulate transcription of the hepcidin encoding hepcidin antimicrobial peptide (HAMP) gene

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