CONTEXT: Baicalin has been characterized as the active compound and quality control marker in Scutellaria baicalensis Georgi, traditionally used as a hypotensive herb. OBJECTIVES: To investigate the inhibitory activities of baicalin against renin and angiotensin-I converting enzyme (ACE) and their molecule mechanism of interactions. METHODS: The fluorescence method using renin substrate 1(R-2932) and the spectroscopy method by Cushman were used to determine renin and ACE activities, respectively. The fluorescence quench techniques were used to characterize their interactions. RESULTS: The results showed that baicalin inhibited renin activity with an IC(50) value of 120.36 µM and inhibited ACE activity with an IC(50) value of 2.24 mM in vitro. The fluorescence emission of both renin and ACE were efficiently quenched by baicalin and a complete quenching was achieved at a high concentration of baicalin. Furthermore, baicalin was more effective in quenching the fluorescence of renin (K(SV) = 60 × 10(3) M(-1)) than ACE (K(SV) = 17.1 × 10(3) M(-1)). The quenching of fluorescence of renin and ACE involved static interactions, which was characterized by the formation of quencher-enzyme complex. The baicalin-renin complex formed through three-sites binding including the active site with a binding constant of 796.15 × 10(13) M(-1), but there was only one binding site for the baicalin-ACE complex with a much smaller binding constant of 6.8 × 10(5) M(-1). CONCLUSION: The inhibition activity of baicalin against renin was a result of the formation of stable complex through multisites binding including the active site, which could explain the higher inhibitory efficiency.
CONTEXT: Baicalin has been characterized as the active compound and quality control marker in Scutellaria baicalensis Georgi, traditionally used as a hypotensive herb. OBJECTIVES: To investigate the inhibitory activities of baicalin against renin and angiotensin-I converting enzyme (ACE) and their molecule mechanism of interactions. METHODS: The fluorescence method using renin substrate 1(R-2932) and the spectroscopy method by Cushman were used to determine renin and ACE activities, respectively. The fluorescence quench techniques were used to characterize their interactions. RESULTS: The results showed that baicalin inhibited renin activity with an IC(50) value of 120.36 µM and inhibited ACE activity with an IC(50) value of 2.24 mM in vitro. The fluorescence emission of both renin and ACE were efficiently quenched by baicalin and a complete quenching was achieved at a high concentration of baicalin. Furthermore, baicalin was more effective in quenching the fluorescence of renin (K(SV) = 60 × 10(3) M(-1)) than ACE (K(SV) = 17.1 × 10(3) M(-1)). The quenching of fluorescence of renin and ACE involved static interactions, which was characterized by the formation of quencher-enzyme complex. The baicalin-renin complex formed through three-sites binding including the active site with a binding constant of 796.15 × 10(13) M(-1), but there was only one binding site for the baicalin-ACE complex with a much smaller binding constant of 6.8 × 10(5) M(-1). CONCLUSION: The inhibition activity of baicalin against renin was a result of the formation of stable complex through multisites binding including the active site, which could explain the higher inhibitory efficiency.