The current treatments of chronic hepatitis B (CHB) face a limited choice of vaccine, antibody and antiviral agents. The development of additional antiviral agents is still needed for improvement of CHB therapy. In this study, we established a screening system in order to identify compounds inhibiting the core promoter activity of hepatitis B virus (HBV). We prepared 80 extracts of marine organisms from the coral reefs of Indonesia and screened them by using this system. Eventually, two extracts showed high inhibitory activity (>95%) and low cytotoxicity (66% to 77%). Solvent fractionation, column chromatography and NMR analysis revealed that 3,5-dibromo-2-(2,4-dibromophenoxy)-phenol (compound 1) and 3,4,5-tribromo-2-(2,4-dibromophenoxy)-phenol (compound 2), which are classified as polybrominated diphenyl ethers (PBDEs), were identified as anti-HBV agents in the extracts. Compounds 1 and 2 inhibited HBV core promoter activity as well as HBV production from HepG2.2.15.7 cells in a dose-dependent manner. The EC₅₀ values of compounds 1 and 2 were 0.23 and 0.80 µM, respectively, while selectivity indexes of compound 1 and 2 were 18.2 and 12.8, respectively. These results suggest that our cell-based HBV core promoter assay system is useful to determine anti-HBV compounds, and that two PBDE compounds are expected to be candidates of lead compounds for the development of anti-HBV drugs.

Secondary metabolites of actinomycetes are a potential source of bioactive compounds in the agricultural sector. This study aimed to determine the fungicidal properties of extracts of marine organism-derived actinomycetes. Actinomycetes were isolated from marine organisms using agar media with 1% colloidal chitin in artificial seawater. Then, the isolates were cultured on liquid media with 1% colloidal chitin in artificial seawater under static conditions for 14 days. The culture was extracted, the fungicide properties were evaluated using the microtiter 96-well plate method, and the influence of inhibition was visualized using apotome and SEM. Finally, the active extract was analyzed using LCMSMS. In the present study, 19 actinomycetes were isolated from marine organisms, and the isolates were examined with regard to their antifungal activities. Of these nineteen isolates, the isolate 19C38A1 was picked out from the rest. Hence, it showed significant control towards F. oxysporum. The prospective strain 19C38A1 was determined to be Kocuria palustris 19C38A1. The extract 19C38A1 was shown to cause damage to cell integrity, indicated by the shrinking form, and inhibited germination in the F. oxysporum; subsequently, the chemical characteristics of the compound produced by the potential isolate 19C38A1 indicated the presence of benzimidazole compounds in the active fraction of C38BK2FA. These results indicate that actinomycetes derived from marine organisms near the coast of Oluhuta, Tomini Bay, Gorontalo, related to strain 19C38A1, are not widely known as sources of valuable fungicides. This preliminary information is important, as it can be used as a basis for further development in the search for fungicides derived from marine actinomycetes.

Hypoxia-adapted cancer cells in tumors contribute to the pathological progression of cancer. The marine spongean sesquiterpene phenols dictyoceratin-A (1) and -C (2) have been shown to induce hypoxia-selective growth inhibition in cultured cancer cells and exhibit in vivo antitumor effects. These compounds inhibit the accumulation of hypoxia-inducible factor-1α (HIF-1α), which is a drug target in hypoxia-adapted cancer cells, under hypoxic conditions. However, the target molecules of compounds 1 and 2, which are responsible for decreasing HIF-1α expression under hypoxic conditions, remain unclear. In this study, we synthesized probe molecules for compounds 1 and 2 to identify their target molecules and found that both compounds bind to RNA polymerase II-associated protein 3 (RPAP3), which is a component of the R2TP/Prefoldin-like (PEDL) complex. In addition, RPAP3-knockdown cells showed a phenotype similar to that of compound-treated cells.

Marine actinomycetes are prolific microorganisms; however, knowledge of their diversity, distribution, and secondary metabolites is limited. Marine actinomycetes represent an untapped source of novel bioactive compounds. In this study, we investigated shrimp shell as substrates for model production bioactive metabolites from actinomycetes under solid state fermentation (SSF) conditions. A total of fifteen actinomycetes were isolated from six sponges and one tunicate. The isolated actinomycetes were grown on solid shrimp shells. Cultures of actinomycetes were extracted with ethyl acetate (EtOAc) and extracts were bioassayed for activity against Staphylococcus aureus. One isolate 18A13O1 from the sponge, Rhabdastrella globostellata, exhibited antibacterial activity on primary screening compared to the other samples and was chosen for further study. Visualization using SEM showed aerial and substrate mycelia. Through phylogenetic analysis, it was confirmed that isolate 18A13O1 is a Pseudonocardia carboxydivorans. Purification of an EtOAc extract yielded A13B2, which showed a minimum inhibition concentration against S. aureus at 15.6 μg/mL. It can be concluded that this basic information is very important for further studies related to the development of the production of bioactive secondary metabolites through the solid state fermentation process.

Shrimp shell waste is a potential source of the biopolymer chitin. Through fermentation, chitin can be converted into its derivative products. This study aimed to isolate and characterize the products of the biodegradation of chitin from shrimp shell waste through a solid-state fermentation process using actinomycetes. Actinomycete isolates were obtained from tunicate marine biota collected from the waters of Buleleng, Bali, using a dilution technique on 1% chitin colloid agar medium. The isolated actinomycetes were cultivated on a shrimp shell waste medium for 7 days, and then the products of the biodegradation of the oligomers were extracted using water. The extracts of the biodegradation products of the shrimp shells were isolated through several chromatographic steps and analyzed using LC–MS–MS, and the bioactivity of the biodegradation products against fungi was tested. The morphological observations and phylogenetic analysis showed that the isolate 18D36-A1 was a rare actinomycete with the proposed name Pseudonocardia antitumoralis 18D36-A1. The results of the analysis using TLC showed that the solid-state fermented water isolate 18D36-A1 produced several oligomeric components. These results indicate that the isolate 18D36-A1 was able to convert chitin into chitooligosaccharides. Further isolation of the extract produced the active fraction D36A1C38, which can inhibit the growth of fungi by 74% at a concentration of 1 mg/mL. This initial information is very important for further studies related to the development of a solid-state fermentation process for obtaining bioactive compounds from shrimp shell waste.

The tumor microenvironment is a nutrient-deficient region that alters the cancer cell phenotype to aggravate cancer pathology. The ability of cancer cells to tolerate nutrient starvation is referred to as austerity. Compounds that preferentially target cancer cells growing under nutrient-deficient conditions are being employed in anti-austerity approaches in anticancer drug discovery. Therefore, in this study, we investigated physcion (1) and 2-(2′,3-epoxy-1′,3′,5′-heptatrienyl)-6-hydroxy-5-(3-methyl-2-butenyl) benzaldehyde (2) obtained from a culture extract of the marine-derived fungus Aspergillus species (sp.), which were isolated from an unidentified marine sponge, as anti-austerity agents. The chemical structures of 1 and 2 were determined via spectroscopic analysis and comparison with authentic spectral data. Compounds 1 and 2 exhibited selective cytotoxicity against human pancreatic carcinoma PANC-1 cells cultured under glucose-deficient conditions, with IC₅₀ values of 6.0 and 1.7 µM, respectively. Compound 2 showed higher selective growth-inhibitory activity (505-fold higher) under glucose-deficient conditions than under general culture conditions. Further analysis of the mechanism underlying the anti-austerity activity of compounds 1 and 2 against glucose-starved PANC-1 cells suggested that they inhibited the mitochondrial electron transport chain.