Matrix metalloproteinase9 (MMP9) is known to be highly expressed during metastatic cancer where most known potential inhibitors failed in the clinical trials. This study aims to select local plants in our state, as anti-breast cancer agent with hemopexin-like domain of MMP9 (PEX9) as the selective protein target. In silico screening for PEX9 inhibitors was performed from our in house-natural compound database to identify the plants. The selected plants were extracted using methanol and then a step-by-step in vitro screening against MMP9 was performed from its crude extract, partitions until fractions using FRET-based assay. The partitions were obtained by performing liquid–liquid extraction on the methanol extract using n-hexane, ethylacetate, n-butanol, and water representing nonpolar to polar solvents. The fractions were made from the selected partition, which demonstrated the best inhibition percentage toward MMP9, using column chromatography. Of the 200 compounds screened, 20 compounds that scored the binding affinity −11.2 to −8.1 kcal/mol toward PEX9 were selected as top hits. The binding of these hits were thoroughly investigated and linked to the plants which they were reported to be isolated from. Six of the eight crude extracts demonstrated inhibition toward MMP9 with the IC₅₀ 24 to 823 µg/mL. The partitions (1 mg/mL) of Ageratum conyzoides aerial parts and Ixora coccinea leaves showed inhibition 94% and 96%, whereas their fractions showed IC₅₀ 43 and 116 µg/mL, respectively toward MMP9. Using MTT assay, the crude extract of Ageratum exhibited IC₅₀ 22 and 229 µg/mL against 4T1 and T47D cell proliferations, respectively with a high safety index concluding its potential anti-breast cancer from herbal.

In our previous work, the partitions (1 mg/mL) of Ageratum conyzoides (AC) aerial parts and Ixora coccinea (IC) leaves showed inhibitions of 94% and 96%, respectively, whereas their fractions showed IC₅₀ 43 and 116 µg/mL, respectively, toward Matrix Metalloproteinase9 (MMP9), an enzyme that catalyzes a proteolysis of extracellular matrix. In this present study, we performed IC₅₀ determinations for AC n-hexane, IC n-hexane, and IC ethylacetate partitions, followed by the cytotoxicity study of individual partitions against MDA-MB-231, 4T1, T47D, MCF7, and Vero cell lines. Successive fractionations from AC n-hexane and IC ethylacetate partitions led to the isolation of two compounds, oxytetracycline (OTC) and dioctyl phthalate (DOP). The result showed that AC n-hexane, IC n-hexane, and IC ethylacetate partitions inhibit MMP9 with their respective IC₅₀ as follows: 246.1 µg/mL, 5.66 µg/mL, and 2.75 × 10⁻² µg/mL. Toward MDA-MB-231, 4T1, T47D, and MCF7, AC n-hexane demonstrated IC₅₀ 2.05, 265, 109.70, and 2.11 µg/mL, respectively, whereas IC ethylacetate showed IC₅₀ 1.92, 57.5, 371.5, and 2.01 µg/mL, respectively. The inhibitions toward MMP9 by OTC were indicated by its IC₅₀ 18.69 µM, whereas DOP was inactive. A molecular docking study suggested that OTC prefers to bind to PEX9 rather than its catalytic domain. Against 4T1, OTC showed inhibition with IC₅₀ 414.20 µM. In conclusion, this study furtherly supports the previous finding that AC and IC are two herbals with potential to be developed as triple-negative anti-breast cancer agents.

In our previous study, aurone compound were found in the n-butanol fraction of Sterculia quadrifida leaves. The aurone compound (compound 1) had high cytotoxic activity in cell lines with IC50 values of 4.05, 12.53, 15.38, and 25.91 μg/mL in 4T1, MCF7, MDA-MB-435, T47D, respectively. In this study, we performed the MMP-9 enzyme inhibition test with FRET assay, and to deepen the mechanism of action of the compounds use molecular docking and molecular dynamics. The results showed that compound 1 had the MMP-9 enzyme inhibition of 91.23% and IC50 48.27%. Furthermore, molecular docking and molecular dynamic simulation suggest compound 1 interacts firmly and stably at the enzyme catalytic site. In conclusion, this study can provide important information on the development of herbal medicines that are potentially cancer drugs.

Context: Medicinal plants have been reported to produce various bioactive molecules. In this study an endophytic fungus was isolated from the leaves of Coleus amboinicus Lour., a medicinal plant used in Jamu and the fungus was identified as Athelia rolfsii. Aims: To isolate, identify and characterize bioactive compound present in ethyl acetate extract of fermentation broth and determine its antimicrobial activities. Methods: The compound was isolated and purified by Preparative Thin Layer Chromatography. Antimicrobial activities were conducted by determining its IC50 and MBC (Minimum Bactericidal Concentration) values against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, Salmonella typhi and Staphylococcus mutans. The structure of the bioactive compound was deduced by spectroscopic data. Results: The bioactive compound displayed antimicrobial activities with IC50 of 0.86, 1.35, 1.33, 2.69, 1.9, 0.24 µg/mL and MBC values of 40, 40, 40, 40, 20, 20 µg/mL against S. aureus, E. coli, P. aeruginosa, B. subtilis, S. typhi, S. mutans, respectively. Based on IR, LC-MS, 13C-NMR, 1H-NMR data, the bioactive compound was suggested to consist of two compounds, methyl hemiterpenoate as a major compound and methyl, 2,3 diene-butanoate as the minor one. Conclusions: The endophytic fungus isolated from the leaves of C. amboinicus produced antimicrobial agents which could be potential for further development.

Breast cancer is one of the most commonly diagnosed cancers and the leading causes of cancer death in women worldwide. Endophytic fungi becomes anticancer metabolite sources which alternatively in plants. The ethyl acetate extract from RL1 code of endophytic fungi from Phyllanthus niruri Linn has cytotoxic activity to T47D cells in vitro. This research objective is to discover active constituent of anticancer which selective in the ethyl acetate extract through cell cycle modulation of T47D cell. Identification of RL1 code endophytic fungi morphology was determined by using microscope. Secondary metabolite production was carried out through fermentation using PDB as medium during two weeks and the medium was extracted with ethyl acetate. Separate and purify method was done using TLC, TLC preparative, and liquid chromatography. Active isolate was characterized by IR, LC-MS, ¹H NMR, ¹³C-NMR, and DEPT. In cytotoxic assay, T47D and Vero cells were cultured in the presence of pure isolate and was evaluated by MTT assay, whereas IC₅₀ and Selectivity Index were used as the parameters to evaluate effectivity of anticancer. Cell cycle distribution was determined by flow cytometry and then analysed by using ModFit LT 3.0 program. Based on microscopic analysis, RL1 code of endophytic fungi is Aspergillus sp. The infrared, mass spectra, ¹H-NMR, ¹³C-NMR, and DEPT signals confirm that isolate 4 is N-(3'-chloro-5'-oxobutyl)-1-methyl-5-phenyl-1H-pyrrole3-carboxamide. Isolate 4 have dose-dependent cytotoxic activity to T47D and Vero cells with their IC₅₀ values of 8,3 and 124 μg/mL, respectively and Selectivity Index is 12,2. Isolate 4 at concentration of 0,9 μg/mL inhibit T47D cell cycle in S-phase. Discovering biosynthetic pathways and molecular mechanisms of cell cycle arrest in isolate 4 still needs to be further investigated.