AKTIVITAS DAN MEKANISME SITOTOKSIK SENYAWA DARI SPONS LAUT Cinachyrella anomala TERHADAP SEL HeLa, T47D, WiDr DAN Vero

AWIK PUJI DYAH NURHAYATI, Dr. Rarastoeti Pratiwi, MSc.

2015 | Disertasi | S3 Ilmu Biologi

Usaha pencarian obat baru dari bahan alam untuk antikanker masih terus dilakukan. Spons laut merupakan bahan alam yang unik memiliki jenis senyawa yang tidak terdapat dalam bahan alami lainnya, oleh karena itu pencarian obat kanker dalam penelitian ini dilakukan pada spons laut. Penelitian ini ditujukan untuk mengetahui sitotoksisitas senyawa dari spons laut Cinachyrella anomala terhadap sel target. Penelitian dilakukan melalui pendekatan Bioassay Guided Isolation sehingga diperoleh senyawa yang dapat digunakan sebagai model pengembangan obat antikanker baru. Sampel spons diekstraksi dengan metode triturasi, fraksinasi, isolasi dan elusidasi senyawa aktif. Pada uji sitotoksisitas dengan metode MTT (Microculture Tetrazolium) ekstrak etil asetat C. anomala menunjukkan sitotoksisitas terhadap sel Hela, T47D, WiDr dan Vero sebesar IC50 388,81; 253,55; 448,67 dan 499,27 μg/mL, sedangkan ekstrak yang tidak larut etil asetat/endapan tidak menunjukkan sitotoksisitas terhadap semua jenis sel. Sel target hasil uji sitotoksisitas ekstrak etil asetat C.anomala adalah sel T47D. Fraksi F1, F2, F3, F4, F5 dan F6 memiliki sitotoksisitas sebesar IC50 237,00; 314,83; 93,80; 124,84; 72,87; 396,15 dan doxorubicin 86,67 μg/mL. Fraksi F5.1, F5.2, F5.3 dan F5.4 menunjukkan sitotoksisitas sebesar IC50 77,31; 156,00; 97,76; 332,17 dan doxorubicin 8,09 μg/mL. Fraksi F5.2 dan F5.3 mampu menurunkan laju proliferasi sel T47D pada inkubasi 24 dan 48 jam pada dosis 125 μg/mL. Fraksi F3.2.1 dan F3.2.2 C. anomala menunjukkan sitotoksisitas sebesar IC50 123,11; 159,90 dan doxorubicin 86,61 μg/mL. Isolat F3.2.1 menunjukkan sitotoksisitas yang paling tinggi dan selanjutnya dilakukan identifikasi struktur kimia berdasarkan pengukuran spektroskopi IR, UV, NMR dan EI-MS. Hasil analisis konfirmasi struktur kimia isolat F3.2.1 yakni senyawa turunan cinachyramine dengan rumus molekul C10H13N3O. Nama struktur yang diusulkan adalah 1,4,9-triazatricyclo[7,3,1,0]trideca-3,5(13),10- trien-8-ol (SA2014). Berdasarkan analisis in siliko senyawa ini diduga memiliki aktivitas sebagai inhibitor cdk2 terhadap sel T47D. Kata kunci: C. anomala, sitotoksik, senyawa 1,4,9-triazatricyclo[7,3,1,0]trideca-3,5(13),10- trien-8-ol, sel HeLa, T47D, WiDr dan Vero

Clinical use of natural products, especially terrestrial ones, is not recent as early medical scripts dating back to ancient Egypt describe the usage of plants to treat malignancies. Although marine-derived compounds are the least represented among the natural products, marine life is becoming increasingly exploited as scientists are overcoming limitations related to accessibility of aquatic samples. Marine sponge is a natural material that has unique types of compounds that are not found in other natural materials. This research was conducted through bioassay-guided isolation approach, compound that can be used as a model of the development of new anticancer medicine. This work was taken to investigate the cytotoxicity of compounds present in the ethyl acetate extract from the marine sponge Cinachyrella anomala against to target human tumor cell lines. Sponge samples were extracted by the method of trituration, fractionation, isolation and elucidation. The test cytotoxicity by MTT methods which ethyl acetate extract against HeLa cancer cells, T47D, WiDr and normal Vero cells were IC50 388.81; 253.55; 448.67 and 499.27 μg/mL. The sediment of ethyl acetate extract has not cytotoxicity against all cell types. T47D cells is the target in cytotoxicity test. Cytotoxicity test of fractions F1, F2, F3, F4, F5 and F6 against T47D cells were IC50 237.00; 314.83; 93.80; 124.84; 72.87; 396.15 and doxorubicin 86.67 μg/mL. Test cytotoxicity of fractions F5.1, F5.2, F5.3 and F5.4 were IC50 77.31; 156.00; 97.76; 332.17 and doxorubicin 8.09 μg/mL. Test cytotoxicity of fractions F3.2.1 and F3.2.2 of C. anomala against cells T47D were IC50 123.11; 159.90 and doxorubicin 86.61 μg/mL. The fraction F3.2.1 of C. anomala showed the highest cytotoxicity and further to identify the chemical structure by spectroscopic measurements of IR, UV, NMR and EI-MS. The results of the analysis confirmed the chemical structure of fraction F3.2.1 was the compound derivative cinachyramine with molecular formula C10H13N3O. The name of the proposed structure is 1,4,9- triazatricyclo [7,3,1,0] trideca-3,5 (13), 10-trien-8-ol (SA2014). Based on in-silico analysis, this compound maight be as an inhibitor for Cdk2 protein which is involved in cells T47D proliferation. Keyword: C. anomala, cytotoxic, compound 1,4,9-triazatricyclo[7,3,1,0]trideca-3,5(13),10- trien-8-ol, HeLa, T47D, WiDr and Vero cells.

Kata Kunci : C. anomala, sitotoksik, senyawa 1,4,9-triazatricyclo[7,3,1,0]trideca-3,5(13),10- trien-8-ol, sel HeLa, T47D, WiDr dan Vero