Aktivitas sitotoksik ekstrak larut heksan daun Micromelum minutum, fraksi-fraksi dan isolatnya terhadap sel kanker payudara MCF-7 dan 4T1

NURRAMADHANI A SIDA, Prof. Dr. Ratna Asmah Susidarti, MS., Apt., Prof. Dr. Edy Meiyanto, M.Si., Apt.

2018 | Tesis | MAGISTER ILMU FARMASI

Micromelum minutum merupakan salah satu tanaman yang diketahui memiliki aktivitas sitotoksik terhadap beberapa sel kanker payudara dengan nilai IC50<100 ug/mL. Isolasi, identifikasi senyawa dan aktivitas farmakologi dari M.minutum yang tumbuh di beberapa negara telah dilakukan, dan diketahui adanya variasi struktur senyawa tergantung lokasi tumbuh dan varietasnya. Meskipun demikian, M.minutum yang tumbuh di Indonesia belum diteliti baik secara fitokimia maupun aktivitas farmakologinya. Penelitian ini bertujuan untuk mengetahui kandungan senyawa dalam ekstrak heksan M.minutum yang tumbuh di Taman Nasional Bantimurung, Bulusaraung, Sulawesi Selatan, serta aktivitas sitotoksik ekstrak heksan, fraksi dan isolatnya terhadap sel kanker payudara MCF-7 dan 4T1. Serbuk simplisia daun M.minutum diekstraksi menggunakan heksan. Ekstrak heksan difraksinasi menggunakan teknik kromatografi. Ekstrak, fraksi dan isoalt diuji aktivitas sitotoksiknya terhadap sel kanker payudara MCF-7 dan 4T1. Konfirmasi stuktur senyawa menggunakan spektroskopi LC-MS, GC-MS, IR, H-NMR, C-NMR, DEPT, dan 2D-NMR (COSY, HSQC, dan HMBC). Ekstrak heksan M. minutum memiliki rendamen sebesar 2.65% terhadap bobot simplisia. Ekstrak memiliki aktivitas sitotoksik terhadap sel kanker payudara MCF-7 dan 4T1 dengan IC50 berturut-turut sebesar 142 dan 77 ug/ml. Pemisahan ekstrak heksan dengan KCV menghasilkan sembilan fraksi (F1-F9). Fraksi 2, 3, dan 6 aktif terhadap sel MCF-7 dan 4T1 dengan IC50 fraksi berturut-turut sebesar 34, 84, dan 115 ug/mL pada sel MCF-7, dan nilai IC50 fraksi berturut-turut sebesar 61, 111 dan 89 ug/mL pada sel 4T1. Analisis F2 dan F3 dengan GC-MS menunjukkan bahwa kedua fraksi tersebut mengandung senyawa sesquiterpen beta elemen (35,36%), longiphinolcarveol (19,80%), dan caryophyllene oxide (14,43%). Pemisahan lebih lanjut F6 menghasilkan senyawa 5,7-dihidroksi-3,4',8-trimetoksiflavon (C18H16O7) yang memiliki aktivitas sitotoksik sedang terhadap sel kanker payudara MCF-7 dan 4T1 dengan nilai IC50 berturut-turut sebesar 127 ug/mL (369 uM) dan 78 ug/mL (226 uM). Berdasarkan hasil dapat disimpulkan ekstrak heksan M. minutum yang tumbuh di Taman Nasional Bantimurung, Bulusaraung, Sulawesi Selatan, mengandung senyawa terpenoid yaitu beta elemen, longiphinolcarveol, dan caryophyllene oxide, serta senyawa 5,7-dihidroksi-3,4',8-trimetoksiflavon. Ekstrak heksan M. minutum, fraksi, dan isolatnya memiliki aktivitas sitotoksik terhadap sel MCF-7 dan 4T1.

Micromelum minutum is an herb that known to have cytotoxic activity against some breast cancer cells with IC50 values <100 ug/mL. Isolation, identification of compounds and pharmacological activity of M. minutum from several countries has been done, and it is known that there are structure variations of compound depended on location and its varieties. Nevertheless, M. minutum that grows in Indonesia has not been studied either phytochemically or pharmacologically. This study aimed to know the compounds in M. minutum leaves hexane extract which grow in Bantimurung National Park, Bulusaraung, South of Sulawesi, and evaluated cytotoxic activity of hexane extract, fraction and isolate against breast cancer cells MCF-7 and 4T1. The powder of M.minutum leaves was macerated using hexane. Than, the hexane extract was fractionated using chromatographic techniques. Extract, fractions, and isolate were tested for cytotoxic activity against breast cancer MCF-7 and 4T1 cells. Structure Of compound was identified by using LC-MS spectroscopy, GC-MS, FT- IR, 1H-NMR, 13C-NMR, DEPT 135o, and 2D-NMR (COSY, HSQC, and HMBC). Micromelum minutum hexane extract had % yield about 2.65% of simplicia weight. The extract had cytotoxic activity against MCF-7 and 4T1 breast cancer cells with IC50 142 and 77 ug/mL, respectively. Fractionation of hexane extract with Vacuum Liquid Chromatography (VLC) resulted in nine fractions (F1-F9). Fraction 2, 3, and 6 were active as cytotoxic agent against MCF-7 and 4T1 cells with IC50 34, 84, and 115 ug /mL, respectively on MCF-7 cells, and IC50 61, 111 and 89 ug/mL, respectively in 4T1 cell. GC-MS analysis of F2 and F3 fractions showed that both fractions contain sesquiterpene beta elemene (35,36%), longiphinolcarveol (19,80%), and caryophyllene oxide (14,43%). Further separation of F6 resulted in a 5,7-dihydroxy-3,4',8-trimethoxyflavone (C18H16O7) compound which had medium cytotoxic activity against MCF-7 and 4T1 breast cancer cells with IC50 value about 127 ug/mL (369 uM) and 78 ug/mL (226 uM), respectively. Based on the result, it could be concluded that hexan M. minutum extract which grows in Bantimurung National Park, Bulusaraung, South Sulawesi contains terpenoid compound ie beta element, longiphinolcarveol, and caryophyllene oxide, and compound 5,7-dihydroxy-3,4',8-trimethoxiflavone. The hexane extract of M. minutum, fractions, and its isolates have cytotoxic activity against MCF-7 and 4T1 breast cancer cells

Kata Kunci : Micromelum minutum, beta elemene, longiphinolcarveol, caryophyllene oxide, 5,7-dihydroxy-3,4',8-trimethoxyflavone, cytotoxic activity, MCF-7, and 4T1