Bioguided Fractionation of Local Plants against Matrix Metalloproteinase9 and Its Cytotoxicity against Breast Cancer Cell Models: In Silico and In Vitro Study (Part II).

Maywan Hariono,I Yoga,Pandu Hariyono,Alfonsus Suryodanindro,Thomas Gonzaga,Abraham Harjono,Adi Hermawansyah,Rollando Rollando,Wahyuning Setyani,Zet Parabang,Michael Yanuar,Habibah Wahab,Rifki Febriansah

doi:10.3390/molecules26051464

Abstract

In our previous work, the partitions (1 mg/mL) of Ageratum conyzoides (AC) aerial parts and Ixora coccinea (IC) leaves showed inhibitions of 94% and 96%, respectively, whereas their fractions showed IC50 43 and 116 µg/mL, respectively, toward Matrix Metalloproteinase9 (MMP9), an enzyme that catalyzes a proteolysis of extracellular matrix. In this present study, we performed IC50 determinations for AC n-hexane, IC n-hexane, and IC ethylacetate partitions, followed by the cytotoxicity study of individual partitions against MDA-MB-231, 4T1, T47D, MCF7, and Vero cell lines. Successive fractionations from AC n-hexane and IC ethylacetate partitions led to the isolation of two compounds, oxytetracycline (OTC) and dioctyl phthalate (DOP). The result showed that AC n-hexane, IC n-hexane, and IC ethylacetate partitions inhibit MMP9 with their respective IC50 as follows: 246.1 µg/mL, 5.66 µg/mL, and 2.75 × 10−2 µg/mL. Toward MDA-MB-231, 4T1, T47D, and MCF7, AC n-hexane demonstrated IC50 2.05, 265, 109.70, and 2.11 µg/mL, respectively, whereas IC ethylacetate showed IC50 1.92, 57.5, 371.5, and 2.01 µg/mL, respectively. The inhibitions toward MMP9 by OTC were indicated by its IC50 18.69 µM, whereas DOP was inactive. A molecular docking study suggested that OTC prefers to bind to PEX9 rather than its catalytic domain. Against 4T1, OTC showed inhibition with IC50 414.20 µM. In conclusion, this study furtherly supports the previous finding that AC and IC are two herbals with potential to be developed as triple-negative anti-breast cancer agents.

Highlights

Cancer is the second leading cause of death in the world with 9.6 million deaths in 2018, with one-in-six mortality caused by this disease [1]
The results showed that OTC (−5.64 kcal/mol) had higher free energy of binding (FEB) than dioctyl phthalate (DOP) (−6.73 kcal/mol) while interacting with a Guanosine Triphosphate (GTP)-binding site less associated with its affinity than DOP against cancer, which might be irrelevant based on the in vitro result
The present study further supported our previous report about the potency of Ageratum conyzoides (AC) and Ixora coccinea (IC) as herbals for breast cancer remedies due to their positive action toward matrix metalloproteinase9 (MMP9) and four cancer cells, including triple-negative sub-types (MDA-MB-231 and 4T1) as well as luminal A subtype (T47D and MCF7)

Summary

Introduction

Cancer is the second leading cause of death in the world with 9.6 million deaths in 2018, with one-in-six mortality caused by this disease [1]. In Asia, approximately 8.75 million new cases and 5.5 million deaths were reported in that same year. In our country (Indonesia), almost 350,000 new cases and 208,000 deaths were reported due to cancer [2]. Triple-negative breast cancer has especially attracted more attention due to the high mortality in females [3]. The high expression of the matrix metalloproteinase (MMP9) protein is one of the indicators during diagnosis [4,5]. Drug discovery targeting this protein is desirable, a potent MMP9 inhibitor is usually limited by its adverse reaction to drugs, such as musculoskeletal syndromes [6,7]

Methods

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Discussion

Conclusion