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Cell invasion is a nature of malignant gliomas, demeriting to many efforts of the treatment. Matrix metalloproteinase (MMP) is acknowledged as a key factor in this complicated process. The aim of this study was to investigate whether inhibition of MMP activity in malignant glioma cells could be achieved by a novel agent, BE16627B (BE). Malignant glioma cell lines, U87MG, U251MG, and U373MG, were employed to evaluate inhibitory effect on zymogram, type IV collagenolysis assay, and haptoinvasion assay for 24 h exposure of BE, following preliminary MTT assay to establish non-cytotoxic dose range. MTT assay revealed that doses of 1, 5, 10, or 50 μM were non-cytotoxic. Zymog ram disclosed that expressions of MMP-2 and MMP-9 were diminished in a dose-dependent fashion. Reflecting that type IV collagenolysis assay revealing that the digestion of type IV collagen was significantly depressed along with elevated dose concentration, haptoinvasion also revealed significantly suppressive effect, meanwhile both 1 μM of BE did not give significant effect. Non-cytotoxic level of BE, ranging from 5 to 50 μM, suppressed MMP-2 and MMP-9 mediated cell invasiveness in the malignant glioma cell lines. This is the first report for cytostatic effect of this agent in glioma cells. This study might be highly implicative of BE16627B to be much meriting to support the other treatment against malignant gliomas.