Molekul, Vol. 18. No. 3, November 2023: 396 – 403 396 MOLEKUL eISSN: 2503-0310 Articles https://doi.org/10.20884/1.jm.2023.18.3.7824 Steroids Produced by Endophytic Fungus Lasiodiplodia Theobromae from Aglaia argentea Blume and Their Cytotoxic Activity Against Hela Cervical Cancer Cell Lines Sari Purbaya 1,2* , Desi Harneti 1 , Asri Peni Wulandari 3 , Yeni Mulyani 1 , Azmi Azhari 1 , Aprilia Permata Sari 1 , Unang Supratman 1,4* 1 Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran, Jatinangor 45363, West Java, Indonesia 2 Department of Chemistry, Faculty of Science and Informatics, Universitas Jenderal Achmad Yani, Cimahi 40531, West Java, Indonesia 3 Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran, Jatinangor 45363, West Java, Indonesia 4 Central Laboratory of Universitas Padjadjaran, Jatinangor 45363, West Java, Indonesia *Corresponding author email: unang.supratman@unpad.ac.id Received January 19, 2023; Accepted June 07, 2023; Available online November 20, 2023 ABSTRACT. Endophytes are micro-organism recognised that living beneficial inside the host plant produced secondary metabolites with biological activity such as anticancer, antitumor, antifungal and antibacterial. In this study, we investigated the endophytic fungus Lasiodiplodia theobromae from the root of Aglaia argentea Blume. A. argentea have been phytochemically investigated previously with unique biological activity such as cytotoxic rocaglate derivatives and dammarane-type triterpenoids. In a continuing project on the bioactive compounds from A. argantea, we have explored endophytic fungi isolated from this plant. Three steroids, ergosterol (1), ergosterol peroxide (2) and stigmasterol (3) have been isolated from endophytic fungus, Lasiodiplodia theobromae derived from the root of Aglaia argentea Blume. The steroids were isolated by vacuum chromatography and column chromatography, the chemical structure was established following the analysis of 1D-NMR, 2D-NMR, IR, MS and by comparison with previously reported spectra data. Ergosterol peroxide (2) and stigmasterol (3) were reported for the first time isolated from Lasiodiplodia theobromae endophytic fungus. Cytotoxic activities of the compounds were tested with resazurin assay against HeLa cervical cancer cells, compound 2 displayed strongest cytotoxic activities against HeLa cervical cancer cells with IC50 values of 0.28 μM, while compounds 1 and 3 showed IC50 values of 0.34 μM and 27.32 μM, respectively. Keyword: Aglaia argentea, cytotoxic activity, HeLa, Lasiodiplodia theobromae, steroid INTRODUCTION Endophytes are micro-organism recognised in last century that living mutually beneficial inside the host plant (Arora & Ramawat, 2017). Interaction of endophytic fungi with plants is a symbiosis in the intracellular space of plants without causing damage to their host plants, have received quite a lot attention (Shiono et al., 2013). Endophytic fungi has enormous potential to produce chemical compounds resulting from secondary metabolites that have unique and interesting activities. Many endophytes fungal produced secondary metabolites with biological activity such as anticancer, antitumor, antifungal and antibacterial which strongly inhibit the growth of other microorganisms (Kharwar et al., 2011; Gunatilaka, 2006). In addition, endophytes can produce secondary metabolites that are the same or similar to their host plants (Supratman, et al., 2021). Fungi produced various kind of secondary metabolite, fungi ability to formed new compound can be done by activating silent gene (Brakhage, 2013). One strain many compounds (OSMAC) method is strategy activated silent gene that are not expressed using conventional fermentation, such as controlled temperature, pH, oxygen condition, salinity, addition of enzyme or tryptophan to rice medium (Gao et al., 2020). Using cultivated on fruit and vegetable juice- supplemented solid rice media is one of OSMAC method for increase diversity of secondary metabolite that not produced when the fungi was grown on rice medium without either fruit or vegetable (Hemphill et al., 2017; Supratman, et al., 2021). Cyclopenta[b]benzofurans are mainly accumulated in the roots and stem bark from Aglaia species (Brader et al., 1998). In this study, we investigated the endophytic fungus Lasiodiplodia theobromae derived from the root of Aglaia argentea Blume, belong to Meliaceae family. We isolated three steroid compounds, ergosterol (1), ergosterol peroxide (2) and stigmasterol (3) using OSMAC method, added