ß-Glucuronidase- and OATP2B1-mediated drug interaction of scutellarin in Dengzhan Xixin Injection: A formulation aspect.

Scutellarin is the major and active constituent of Dengzhan Xixin Injection (DZXX), a traditional Chinese medicine prepared from the aqueous extract of Erigeron breviscapus and widely used for the treatment of various cerebrovascular diseases in clinic. In present study, the possible pharmacokinetic differences of scutellarin after intravenous administration of scutellarin alone or DZXX were explored. Additional, the potential roles of ß-glucuronidase (GLU) and OATP2B1 in drug-drug interaction (DDI) between scutellarin and constituents of DZXX were further evaluated in vitro. The plasma concentration, urinary and biliary excretion of scutellarin in rats after administration of DZXX, were significantly higher than those received scutellarin, while pharmacokinetic profile of Apigenin 7-O-glucuronide (AG) in rats was similar no matter AG or DZXX group. Furthermore, higher concentration in brain and plasma, however, lower level of scutellarin in intestine were observed after intravenous administration of DZXX. Finally, AG and caffeoylquinic acid esters were found to significantly inhibit GLU and OATP2B1 in vitro, which might explain, at least in part, the pharmacokinetic DDI between scutellarin and other chemical constituents in DZXX. The findings provided deep insight into the prescription-formulating principle in DZXX for treating the cerebrovascular diseases.

Mechanistic studies on the absorption and disposition of scutellarin in humans: selective OATP2B1-mediated hepatic uptake is a likely key determinant for its unique pharmacokinetic characteristics.

Scutellarin [scutellarein-7-O-glucuronide (S-7-G)] displayed a unique pharmacokinetic profile in humans after oral administration: the original compound was hardly detected, whereas its isomeric metabolite isoscutellarin [scutellarein-6-O-glucuronide (S-6-G)] had a markedly high exposure. Previous rat study revealed that S-7-G and S-6-G in the blood mainly originated from their aglycone in enterocytes, and that the S-7-G/S-6-G ratio declined dramatically because of a higher hepatic elimination of S-7-G. In the present study, metabolite profiling in human excreta demonstrated that the major metabolic pathway for S-6-G and S-7-G was through further glucuronidation. To further understand the cause for the exposure difference between S-7-G and S-6-G in humans, studies were conducted to uncover mechanisms underlying their formation and elimination. In vitro metabolism study suggested that S-7-G was formed more easily but metabolized more slowly in human intestinal and hepatic microsomes. Efflux transporter study showed that S-6-G and S-7-G were good substrates of breast cancer resistance protein and multidrug resistance-associated protein (MRP) 2 and possible substrates of MRP3; however, there was no preference great enough to alter the S-7-G/S-6-G ratio in the blood. Among the major hepatic anion uptake transporters, organic anion-transporting polypeptide (OATP) 2B1 played a predominant role in the hepatic uptake of S-6-G and S-7-G and showed greater preference for S-7-G with higher affinity than S-6-G (K(m) values were 1.77 and 43.9 µM, respectively). Considering the low intrinsic permeability of S-6-G and S-7-G and the role of OATP2B1 in the hepatic clearance of such compounds, the selective hepatic uptake of S-7-G mediated by OATP2B1 is likely a key determinant for the much lower systemic exposure of S-7-G than S-6-G in humans.

Use of hollow fiber liquid phase microextraction and HPLC for extraction and determination of apigenin in human urine after consumption of Satureja sahendica Bornm.

The applicability of hollow fiber liquid phase microextraction (HF-LPME) was evaluated for extraction and preconcentration of apigenin prior to its determination by HPLC. Different parameters affecting the HF-LPME recovery such as nature of organic solvent, pH of donor and acceptor phases, extraction time, stirring speed, salt addition were optimized. Under optimum conditions (1-octanol as organic solvent, pH of the donor phase=3 and pH of acceptor phase=11.5, extraction time of 75 min, stirring speed of 1000 rpm) limit of detection (LOD) of 0.1 ng/mL, linear range of 0.5-300 ng/mL and correlation of determination (R(2)) of 0.9956 were obtained. The relative intra and inter-day standard deviations (RSD%) based on five replicate measurement were 3.5% and 10.7% respectively. Enrichment factor of 315 and recovery 85% were achieved. Finally, the applicability of the proposed method was evaluated by extraction and determination of apigenin in urine sample after consumption of Satureja sahendica Bornm. which is a native medicinal plant from Iran. Concentration of apigenin in urine sample was found to be 6.20 ng/mL.

Two major urinary metabolites of scutellarin in rats.

Urinary metabolites of scutellarin, the main effective constituent of breviscapine, a cerebrovascular and cardiovascular drug consisting of total flavonoids of Erigeron breviscapus, were investigated in rats. Two major metabolites were isolated from the urine of rats following oral administration of scutellarin and identified as scutellarein 6,7-di- O-beta-D-glucuronide (M1) and scutellarein (M2), respectively, on the basis of chemical and spectroscopic evidence. M1 was reported as a metabolite of scutellarin for the first time.