Clinacanthus nutans is a well-known herb that has been used as an alternative and therapeutic medicine, however more selective C. nutans extracts are needed. In this study, leaves were extracted with 80% methanol and further fractionated with n-hexane, dichloromethane, chloroform, n-butanol, and aqueous residue. Subsequently, the total phenolic content (TPC), total flavonoid content (TFC), antioxidant scavenging activity, and antiproliferative effects on breast cancer (Michigan Cancer Foundation-7 [MCF7]) and normal breast (Michigan Cancer Foundation-10A [MCF 10A]) cells of the extracts were measured. Additionally, molecular docking simulation of the major compounds from C. nutans extracts was conducted. The aqueous residue had the highest TPC and TFC, whereas the crude extract had the highest scavenging activity. Among the extracts, dichloromethane extract (CN-Dcm) was selected as it had the highest selectivity index (SI) (1.48). Then, the chosen extract (CN-Dcm) was proceed for further analysis. The compounds from CN-Dcm were identified using gas chromatography–mass spectrometry (GC-MS). The major compounds from CN-Dcm were further investigated through molecular docking studies. Palmitic acid and linolenyl alcohol were the compounds found in the CN-Dcm extract that exhibited the highest binding affinities with p53-binding protein Mdm-2. These results highlight the potential of C. nutans as a source of anticancer activities.

Objective: This study was conducted to investigate the antiproliferative activity of extracts of Clinacanthus nutansleaves against human cervical cancer (HeLa) cells. Methods: C. nutans leaves were subjected to extraction using 80%methanol or water. The methanol extract was further extracted to obtain hexane, dichloromethane (DCM), and aqueousfractions. The antiproliferative activity of the extracts against HeLa cells was determined. The most cytotoxic extractwas furthered analyzed by apoptosis and cell cycle assays, and the phytochemical constituents were screened by gaschromatography-mass spectrometry (GC-MS). Results: All of the extracts were antiproliferative against HeLa cells, andthe DCM fraction had the lowest IC50 value of 70 μg/mL at 48 h. Microscopic studies showed that HeLa cells exposedto the DCM fraction exhibited marked morphological features of apoptosis. The flow cytometry study also confirmedthat the DCM fraction induced apoptosis in HeLa cells, with cell cycle arrest at the S phase. GC-MS analysis revealedthe presence of at least 28 compounds in the DCM fraction, most of which were fatty acids. Conclusion: The DCMfraction obtained using the extraction method described herein had a lower IC50 value than those reported in previousstudies that characterized the anticancer activity of C. nutans against HeLa cells.

Background: Medicinal herbs in Malaysia like Clinacanthus nutans (CN) traditionally are used in the treatment of various diseases and cancers. The present research was conducted to determine the effects of C. nutans leaf different solvent extracts on the human breast cancer cell lines (MCF-7). The antiproliferative growth and survival effects of dichloromethane CN leaf extracts (CNDCM), as well as the short- and long-term effects through metastasis, apoptosis and cell cycle effects, were observed. The chemical profiles were done to evaluate the properties of the CNDCM. Results: The evaluation of GC–MS identified 16 major phytochemical compounds present in this extract with biological activities. Antiproliferative assay used is the SRB assay, which showed the CNDCM induced strong antiproliferative property compared with the other extracts, so its IC₅₀ dose was selected for further testing with value 108 µg/mL at 72 h after exposure on MCF-7 and MCF-10A cell lines. The clonogenic survival effects of CNDCM in various concentrations (31.25, 62.5, 125, 250 and 500 µg/mL) inhibited the ability of MCF-7 cells to form colonies, and the metastasis result was indicated in an image of wound healing assay. Moreover, the CNDCM extract significantly induced apoptosis in all the cell cycle phases. Finally, the experiments with various extract concentrations on normal human breast cell lines showed no antiproliferative effects for all the extracts tested. Conclusion: Among all the extracts of CN, the CNDCM extracts demonstrated the highest antiproliferative activity and survival against the MCF-7 cell lines tested.