Sintesis dan Epoksidasi N,N-dietiloleamida dan N-propiloleamida: Evaluasi Sitotoksisitas terhadap Sel HeLa, T47D, dan Vero serta Studi Interaksi Molekuler dengan Protein Fatty Acid Synthase (FASN)
Novia Ariska Pratiwi, Prof. Drs. Jumina, Ph.D.; Dr. Sc. Robby Noor Cahyono, S.Si., M.Sc.
2025 | Tesis | S2 Ilmu Kimia
Indonesia merupakan salah satu penghasil minyak kelapa sawit terbesar di dunia, yang menjadi sumber utama asam oleat dengan potensi aktivitas biologis, termasuk sebagai agen antikanker. Penelitian ini bertujuan untuk mensintesis dan mengevaluasi empat senyawa kandidat antikanker turunan asam oleat yaitu N,N-dietiloleamida, N-propiloleamida, epoksida N,N-dietiloleamida, dan epoksida N-propiloleamida. Asam oleat dapat diperoleh melalui proses hidrolisis FAME (Fatty Acid Methyl Esters) yang telah ditingkatkan kemurniannya melalui UIC (Urea Inclusion Complex). Senyawa amida disintesis melalui amidasi dengan dietilamina dan propilamina menggunakan tionil klorida, lalu diepoksidasi dengan asam performat untuk memperoleh senyawa epoksida. Struktur senyawa dikarakterisasi menggunakan FTIR, GC-MS/LC-MS, ¹H-NMR, dan ¹³C-NMR. Uji sitotoksisitas secara in vitro dilakukan terhadap sel kanker payudara (T47D), sel kanker serviks (HeLa), dan sel normal Vero. Studi penambatan molekul dilakukan terhadap protein FASN subunit tioesterase.
Proses transesterifikasi berhasil menghasilkan FAME dengan persen hasil sebesar 82,90% (b/b) dan kelimpahan relatif metil oleat mencapai 43,50%. Setelah dilakukan pemurnian melalui proses inklusi urea, rendemen menurun menjadi 35,20% (b/b) namun kemurnian metil oleat meningkat menjadi 72,20%. Asam oleat hasil hidrolisis kemudian digunakan sebagai prekursor dalam sintesis N,N-dietiloleamida, N-propiloleamida, epoksida N,N-dietiloleamida, epoksida N-propiloleamida, yang masing-masing diperoleh dengan persen hasil berturut – turut sebesar 91,18% (b/b); 84,65% (b/b); 70,82% (b/b); 81,22% (b/b). Uji sitotoksik menunjukkan senyawa hasil sintesis memiliki aktivitas antikanker yang signifikan terhadap sel kanker T47D dan HeLa dengan nilai IC?? berturut-turut sebesar 22,85; 36,96; 16,27; dan 31,63 µg/mL (T47D), serta 21,07; 92,83; 16,27; dan 22,70 µg.mL (HeLa). Indeks selektivitas beberapa senyawa menunjukkan potensi selektif terhadap sel kanker dibandingkan sel normal Vero dengan nilai berturut-turut 5,41; 6,78; 1,20; 2,05 (T47D), dan 5,86; 2,70; 1,20; 2,86 (HeLa). Studi penambatan molekul menunjukkan afinitas ikatan kuat (?4,26; ?4,22; ?4,68; dan ?4,27 kkal/mol) pada sisi aktif FASN yang mendukung hasil uji in vitro.
Indonesia is one of the world’s largest producers of palm oil, which serves as a primary source of oleic acid with potential biological activities, including as an anticancer agent. This study aims to synthesize and evaluate four oleic acid-derived compounds as potential anticancer candidates, specifically N,N-diethyloleamide, N-propyloleamide, epoxide N,N-diethyloleamide, and epoxide N-propyloleamide. Oleic acid can be obtained through the hydrolysis of FAME (Fatty Acid Methyl Esters), whose purity has been enhanced using the UIC (Urea Inclusion Complex) method. The amide compounds were synthesized via amidation with diethylamine and propylamine using thionyl chloride, followed by epoxidation with performic acid to yield the epoxide derivatives. The structures of the compounds were characterized using FTIR, GC-MS/LC-MS, ¹H-NMR, and ¹³C-NMR. In vitro cytotoxicity assays were conducted against breast cancer cells (T47D), cervical cancer cells (HeLa), and normal Vero cells. Molecular docking studies were performed targeting the thioesterase subunit of the FASN protein.
The transesterification process successfully produced FAME with a yield of 82.90% (w/w), and the relative abundance of methyl oleate reached 43.50%. After purification through urea inclusion, the yield decreased to 35.20% (w/w), but the purity of methyl oleate increased to 72.20%. The oleic acid obtained from hydrolysis was then used as a precursor in the synthesis of N,N-diethyloleamide, N-propyloleamide, epoxide N,N-diethyloleamide, and epoxide N-propyloleamide, which were obtained with yields of 91.18% (w/w); 84.65% (w/w); 70.82% (w/w); and 81.22% (w/w), respectively. Cytotoxicity assays showed that the synthesized compounds exhibited significant anticancer activity against T47D and HeLa cancer cells, with IC?? values of 22.85; 36.96; 16.27; and 31.63 µg/mL (T47D), and 21.07; 92.83; 16.27; and 22.70 µg/mL (HeLa), respectively. The selectivity indices of several compounds indicated potential selectivity towards cancer cells compared to normal Vero cells, with values of 5.41; 6.78; 1.20; and 2.05 (T47D), and 5.86; 2.70; 1.20; and 2.86 (HeLa), respectively. Molecular docking studies revealed strong binding affinities (?4.26; ?4.22; ?4.68; and ?4.27 kcal/mol) at the active site of FASN, supporting the in vitro assay results.
Kata Kunci : minyak kelapa sawit, epoksidasi, amida, antikanker
