Synthesis and Anticancer Activity of (E)-2’-hydroxy-2-bromo-4,5-dimethoxychalcone Against Breast Cancer (MCF-7) Cell Line

Breast cancer is one of cancer causes of death in woman. Chemotherapy is one cancer treatment give toxic effects on normal cells. Alternative of cancer treatment by using flavonoid derivative have potent anticancer to reduce side effects of cancer. Chalcone is family of flavonoid that have biological activity. Chalcone derivatives have the potent as anticancer agent. Chalcone with the presence halogen, metoxy group in ring B is known to inhibit cancer cells. The aims of this research were to synthesize chalcone derivative with bromo, methoxy, and hydroxy groups in chalcone ring and to determine the anticancer activity of chalcone derivative. The chalcone derivative was synthesized from 2hydroxyacetophenone with 2-bromo-4,5-dimethoxybenzaldehyde by Claisen-Schmidt reaction. In vitro cytotoxicity against breast cancer cell was tested by MTT assay method. The compound of (E)-2’-hydroxy-2-bromo-4,5-dimethoxychalcone was yield in 78% as yellow solid. The IC50 of (E)-2’-hydroxy-2-bromo-4,5-dimethoxychalcone was 42.19 μg/mL as a moderate activity to inhibiting breast cancer cell line. Cytotoxity of docorubicin againts breast cancer cell line more active than (E)-2’-hydroxy-2-bromo-4,5-dimethoxychalcone with IC50 10.61 μg/mL. Doxorubicin as drug standar had better anticancer activity than (E)-2’-hydroxy-2-bromo-4,5-dimethoxychalcone. Based on the IC50 value, (E)-2’-hydroxy-2-bromo4,5-dimethoxychalcone has a moderate activity towards breast cancer cell lines. Thus, this compound can be recommended as candidate for anticancer againts breast cancer cell lines. Keyword : anticancer; chalcone derivative; breast cancer INTRODUCTION Cancer is a diseases involving uncontrolled growth of abnormal cell in body. Cancer became the second leading cause of death after cardiovascular disease (Sashidhara, Kumar, Kumar, Sarkar, & Sinha, 2010). Based on the World Health Organization (WHO) estimation, in 2030 the deaths caused by cancer will increase by 44% with respect to 2008. Breast cancer is the most common invasive cancer in female worldwide (Mellado-Garcia, Reyna, Weinstein-Oppenheimer, Cuellar, & Aguilar, 2018). It accounts for 16% of all female cancers and 22.9% of invasive cancers in woman, 18.2% of all cancer deaths worldwide including both in males and females are from breast cancer (Rajak et al., 2010). Breast cancer became the first ranked among other cancers disease cause of death from cancer in woman from hospital data in Indonesia. In 2013, breast cancer is type of cancer with most occured in Indonesia with 61,682 cases (Kementerian Kesehatan, 2015). There are many diagnostic methods and therapeutic approaches for cancer including surgery, radiation, chemotherapy, hormone therapy and biologycal therapy (Kameshwaran, Suresh, Arunachalam, Kanthlal, & Mohanraj, 2012). Nowdays, chemotherapeutic agents is one of the notable healing approaches for several cancer types, e.g breast (Smith, Heutcheon, & Heys, 2000). The problem of chemotherapeutical is able to kill cancer cell but also kill normal cells around (Ketabforoosh, Kheirollahi, Safavi, & Esmati, 2014). Ranjit et al. (2013) reported that chemotherapeutic agents can be used as an alternative cancer treatment but some chemotherapeutic agents give negative effects such as depression and alopecia. Therefore, novel anticancer agents must be developed in the medical chemistry. Natural products have been used to develop new compounds with potential application for several pharmacological treatments (Yu, Shen, Song, & Xie, 2018). Polyphenolic compounds that have been reported to posses anticancer e.g flavonoids and their derivative compounds (Agrawal, 2011). Chalcone, the precursors of flavonoids, are widely present in fruits, vegetables, and are present in plant species (Zaini et al., 2019). Chemically, they consist of open-chain flavonoids in which the two aromatic Synthesis and Anticancer Activity Retno Aliyatul Fikroh, et al. 35 rings are joined by a three-carbon αβ–unsaturated carbonyl system (Syam, Abdelwahab, Al-mamary, & Mohan, 2012). Chalcone have been reported as several pharmacological activity including antiinflammatory (Hsieh, Lee, Wang, Wang, & Lin, 1998), antifungal (Sivakumar, Muthu Kumar, & Doble, 2009), antimalaria (Syahri, Yuanita, Nurohmah, Armunanto, & Purwono, 2017), and anticancer (Bonakdar et al., 2017). Substituents on chalcones have an important role in improving activity og anticancer againts. According to Rao, Fang, & Tzeng (2004) reported that chalcone with methoxy groups on the aromatic ring has potential as an anti-proliferation in tumor cells without giving any toxic effects on normal cells. Sofiana, Balatif, & Zamri (2014) states that chalcone with methoxy groups have a greater toxic properties due to the αβunsaturated bond and mesomeri positive effects from methoxy substituted. Ketabforoosh et al. (2014) have tested the activity chalcone without substituent between 2-bromo hydroxyacetophenone and veratraldehyde with IC50 value of 6.1 μM (1.73 mg/mL) against breast cancer cells MCF-7. The study also synthesized chalcone between 2-hydroxyacetophenone and veratraldehyde with bromo substituted have IC50 value of 1 μM (0.36 mg/mL), it is lower than compound without the bromo substituent that is equal to 1 μM (0.36 mg/mL). This shows that the halogen substituent such as bromo also have activity in inhibiting breast cancer. Chalcone derivatives with halogen substituent such as bromo very useful to modulate the electron and steric characteristics of the drug as well as affecting the balance of the hydrophilic hydrophobic molecules. Syam et al. (2012) have synthesized chalcone with halogen substituent such as cloro at ring B with IC50 value of 7.991 μg/mL and methoxy substituen with IC50 11.625 μg/mL. In this research showed that chalcone with halogen subtituent in ring B increased the ROS level eventually leading to apoptosis in MCF-7 cells through intrinsic as well as extrinsic pathways. According to the available data, we synthesized a novel chalcone derivative by presence halogen and methoxy substituents at ring B of chalcone to find a new anticancer agent. The synthesis of chalcone in this research used simple steps with produced in more than 50% yield. The cytotoxicity was determined by MTT assay toward breast cancer (MCF-7) cell lines. EXPERIMENTAL SECTION Materials The material with pro analysis grade were purchased from Merck, i.e., 2-hydroxyacetophenone, 3,4-dimethoxybenzaldehyde, sodium hydroxide, ethyl ecetate, ethanol, methanol, n-hexane, glacial acetic acid, hydrocloric acid, hydrobromic acid, while sodium sulfate anhydrous and potassium bromate were from Sigma-Aldrich. All reactions were monitored with Thin Layer Chromatography plat of aluminium F254 from Merck were obtained with silica gel 60 (20 x 20 cm). Purification was performed by column chromatography from Merck with silica gel 60. The material for in vitro test were breast cancer cell line (MCF-7), doxorubicin, fetal bovine serum 10% (w/v) from Sigma-Aldrich, Medium Dulbecco’s Modified Eagle’s Medium (DMEM) from SigmaAldrich, dimethyl sulfoxide, Sodium Dedocyl Sulfate (SDS) 10%, Phospate Buffer Solution (PBS), solution of MTT (3-(4,5-dimethythiazol-2-yl)-2,5diphenyltetrazolium-bromide and hydroclorid acid