The Antiviral Effect of Indonesian Medicinal Plant Extracts Against Dengue Virus In Vitro and In Silico.

Dengue infections are still a worldwide burden, especially in Indonesia. There is no specific medication against the dengue virus. Recently, many types of research have been conducted to discover a new drug for dengue virus using natural resource extracts. Indonesia, as a tropical country, has a wide biodiversity. There are several medicinal plants in Indonesia that are believed to possess anti-dengue activity, such as Myristica fatua, Cymbopogon citratus, and Acorus calamus plants. We conducted an in vitro laboratory experiment of several extracts from Indonesian herbs combined with in silico analysis. The extracts were evaluated for safety and antiviral activity in Huh7it-1 cell lines, using a single dose of 20 µg/mL and dose-dependent (5, 10, 20, 40, 80 and 160 µg/mL) of plant extracts against dengue virus serotype 2 (DENV-2) NGC strain. The DMSO 0.1% was used as a negative control. The cytotoxic aspect was assessed by counting the cell viability, while the antiviral activity was calculated by counting the average inhibition. The selectivity index (SI) of plant extracts were performed from a ratio of CC50/EC50 value. In silico analysis was conducted to determine the free energy of binding between NS5 of dengue virus with bioactive compounds contained in Myristica fatua, Cymbopogon citratus and Acorus calamus extract plants. We determined that all extracts were not toxic against Huh7it-1 cell lines. The methanolic extracts of A. calamus, C. citratus, and M. fatua showed inhibition of DENV-2 at a dose of 20 µg/mL to 96.5%, 98.9%, and 122.7%, respectively. The dose-dependent effects showed that M. fatua has the best inhibition activity towards DENV-2. Molecular docking result showed that artesunic acid within M. fatua has the best free energy of binding (-7.2 kcal/mol), followed by homoegonol (-7.1 kcal/mol) which was slightly different from artesunic acid among others. The methanolic extracts of A. calamus, C. citratus, and M. fatua showed prospective anti-dengue activities both in vitro and in silico. Future research should be conducted to find the pure extracts of all useful herbs as a new candidate of antiviral drug.

(+)-Altholactone exhibits broad spectrum immune modulating activity by inhibiting the activation of pro-inflammatory cytokines in RAW 264.7 cell lines.

An evaluation of Indonesian plants to identify compounds with immune modulating activity revealed that the methanolic extract of an Alphonsea javanica Scheff specimen possessed selective anti-inflammatory activity in a nuclear factor-kappa B (NF-κB) luciferase and MTT assay using transfected macrophage immune (Raw264.7) cells. A high-throughput LC/MS-ELSD based library approach of the extract in combination with the NF-κB and MTT assays revealed the styryl lactone (+)-altholactone (2) was responsible for the activity. Compound 2, its acetylated derivate (+)-3-O-acetylaltholactone (3), and the major compound of this class, (+)-goniothalmin (1), were further evaluated to determine their anti-inflammatory potential in the NF-κB assay. Concentration-response studies of 1-3 indicated that only 2 possessed NF-κB based anti-inflammatory activity. Compound 2 reduced the LPS-induced NO production, phosphorylation of IκBα, and the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) using Western blot analysis. Further studies using qPCR indicated 2 reduced the expression of eight pro-inflammatory cytokines/enzymes (0.8-5.0µM) which included: COX-2, iNOS, IP-10, IL-1ß, MCP-1, GCS-F, IL-6 and IFN-ß. These results indicated that 2 displays broad spectrum immune modulating activity by functioning as an anti-inflammatory agent against LPS-induced NF-κB signaling. Conversely the selective cytotoxicity and in vivo anti-tumor and anti-inflammatory activity previously reported for 1 do not appear to arise from a mechanism that is linked to the NF-κB immune mediated pathway.

Identification of cytotoxic compound from Artocarpus communis leaves against P-388 cells.

In the course of continuing research for finding bioactive compounds from Indonesian plants, the leaves of Artocarpus communis was extracted by ethanol. This extract partitioned with n-hexane-water (1:4) and then water extract was partitioned with dichloromethane. Dichloromethane extract was purified by column chromatography techniques on silica gel to afford yellow crystal (F-1). Based on LC-MS, 1H-NMR and 13C-NMR (1D and 2D) spectra and compared with previous spectral data, it was identified as prenylated flavonoid, 1-(2,4-dihydroxyphenyl)-3-[8-hydroxy-2-methyl-2-(4-methyl-3-pentenyl)-2H-1-benzopyran-5-yl] 1-propanone. This compound showed significant cytotoxicity against murine P-388 leukemia cells.

Antioxidant compound from the rhizomes of Kaempferia rotunda L.

This research aimed to investigate the antioxidant effect from rhizomes of K. rotunda for finding the active compounds by DPPH free radical scavenging activity assay. The chloroform-soluble extract of the rhizomes of K. rotunda showed significant scavenging effect on the on 1,1-diphenyl-2-picryl hydrazyl (DPPH) free radicals (IC50 = 180 microg mL(-1)). Two compounds of the chloroform-soluble extract were isolated and identified. Compound 1, 2'-hydroxy-4,4',6'-trimethoxy-chalcone was found as the active constituent (IC50 = 142 microg mL(-1)). Compound 2, (+)-crotepoxide, was inactive (IC50 = 1516 microg mL(-1)). The structures of compounds 1 and 2 were identified based on the basis of spectral evidence, Mass Spectrophotometry (MS) and 2D-NMR (2 dimension of Nuclear Magnetic Resonance) data including Heteromolecular Multiple Quantum Coherence (HMQC) and Heteromolecular Multiple Bond Correlation (HMBC) and comparison to published values.