Antimetastatic and Antitumor Activities of Orally Administered NAX014 Compound in a Murine Model of HER2-Positive Breast Cancer.

Mauro Provinciali,Gaetano Fiorillo,Francesco Piacenza,Paolo Lombardi,Cristina Geroni,Carmela Salvatore,Fiorenza Orlando,Elisa Pierpaoli

doi:10.3390/ijms22052653

Abstract

The natural isoquinoline alkaloid Berberine (BBR) has been shown to possess several therapeutic effects, including anticancer activity. Different BBR derivatives have been designed and synthesized in order to obtain new compounds with enhanced anticancer efficacy. We previously showed that intraperitoneal (IP) administration of the BBR-derived NAX014 compound was able to counteract HER-2 overexpressing mammary tumors onset and progression in transgenic mice. However, the IP administration was found to induce organ toxicity at doses higher than 2.5 mg/Kg. In this study, we evaluated the effect of intragastric (IG) administration of 20 mg/kg of NAX014 on both safety and anticancer efficacy in HER-2/neu transgenic mice. Furthermore, cancer cell dissemination and migration, tumor cell senescence and immunological changes were examined. Our results demonstrated that IG NAX014 administration delayed the onset of mammary tumors with no negative effects on health and survival. NAX014 reduced HER-2 overexpressing BC cells migration in vitro and the frequency of lung metastasis in HER-2/neu transgenic mice. A statistically significant increase of senescence-associated p16 expression was observed in tumors from NAX014-treated mice, and the induction of cell senescence was observed in HER-2 overexpressing BC cells after in vitro treatment with NAX014. Although NAX014 did not modulate the presence of tumor-infiltrating lymphocytes, the level of circulating TNF-α and VEGF was found to be reduced in NAX014-treated mice. The overall results address the NAX014 compound as potential tool for therapeutic strategies against HER-2 overexpressing breast cancer.

Highlights

Breast cancer (BC) is the most frequent cancer in women worldwide and the second leading cause of cancer-related deaths in women
We found in an in vitro model that the presence of the electron-withdrawing group Cl on the aromatic moiety linked to NAX014 was able to exert a higher anti-proliferative effect on human epidermal growth factor receptor 2 (HER-2) overexpressing Breast Cancer (BC) cells with respect to that one brought by BBR [8]
We evaluated the effect of IG administration of the BBR-derived NAX014 compound (Figure 1A) on spontaneous mammary carcinogenesis in HER-2/neu transgenic mice

Summary

Introduction

Breast cancer (BC) is the most frequent cancer in women worldwide and the second leading cause of cancer-related deaths in women. Intraperitoneal (IP) administration of NAX014 was able (a) to delay the development of mammary tumors, (b) to reduce the tumor volume and tumor vessel density and (c) to increase tumor cell senescence in HER-2/neu transgenic mice [11]. In this experimental model, the dosage of NAX014, and BBR in a higher extent, was limited because of increasing organ injury and mortality rate with doses higher than 2.5 mg/Kg, as observed during the toxicity study [11]. The effect of NAX014 on cell migration/invasion and induction of cell senescence was evaluated both in vitro and in vivo confirming their role in the inhibition of tumorigenesis

Results

Chemicals

Cell lines and Treatments

Experimental Design

Flow Cytometric β-Galactosidase Assay

Statistical Analysis