However, it is unclear which ingredients in TZQ are responsible for the -glycosidase inhibitor activity. disappearance of -sheets and an increase in the -helix content of the enzyme, similar to acarbose. Conclusions This work provides useful PF-06305591 information for the inhibitory effect of TZQ on maltase. TZQ has the potential to be an -glycosidase inhibitor for the prevention and treatment of prediabetes or mild diabetes mellitus. Background Elevated postprandial glucose, which is one of the earliest abnormalities of glucose homeostasis associated with diabetes, will increase the risk of developing microvascular complications and cardiovascular disease [1, 2]. Postprandial glycaemia often accompanies several long-term complications, such as nephropathy, hypertension, atherosclerosis and hyperlipidaemia . Diabetes mellitus is becoming a serious threat to human health worldwide . One encouraging approach for better control of postprandial glycaemia is to reduce carbohydrate digestion . The clinically used -glucosidase inhibitors, including acarbose, miglitol and voglibose, can bind to -glucosidase and competitively inhibit the enzyme in the small intestine to delay the expeditious generation of blood glucose [1, 6]. As a Chinese herbal medicine, Tangzhiqing (TZQ) is composed of Gaertn. leaves, Pall. roots, roots, L. leaves, and leaves and has a long history of use in treating diabetes mellitus. Based on the recipe of TZQ, we developed a new formula including eight fractions of red paeony saponins, lotus leaf alkaloids, lotus leaf flavonoids, mulberry leaf alkaloids, mulberry leaf flavonoids, mulberry leaf polysaccharide, danshen polyphenols, and hawthorn leaf flavonoids with a ratio of 3.0:2.9:1.8:0.8:0.1:14.0:0.8:0.2 (w/w) . The fractions of mulberry leaf alkaloids, mulberry leaf flavonoids, and hawthorn leaf flavonoids significantly inhibited glucose absorption. The TZQ formula possesses the effects of anti-hyperlipidaemia, anti-hyperglycaemia, and anti-oxidative PF-06305591 stress, which suggests that TZQ could be developed as a potential ready-made formula for pre-diabetes treatment [7C9]. Moreover, the TZQ formula was used in an Investigational New Drug Application study by the China Food and Drug Administration in November 2010. Preclinical studies have shown that TZQ has obvious inhibitory effects on rat intestinal saccharase for sucrase and maltase in vivo . In genetically modified KK-Ay mice with type 2 diabetes, TZQ presented beneficial effects on the improvement of glucose metabolism by reducing -glycosidase activity . TZQ has the same effects as acarbose, which inhibits the postprandial increase in blood glucose levels by inhibiting and delaying digestion and absorption of carbohydrates in healthy Chinese volunteers. Moreover, TZQ was found to significantly regulate abnormal glucose, decrease insulin secretion to maintain PF-06305591 normoglycaemia, and reduce glycosylated haemoglobin (HbA1c) and fasting insulin in type 2 diabetes mellitus patients [10, 11]. An eight-period, self-crossover clinical trial in healthy volunteers was performed to determine the effect of TZQ on the glycaemic index (GI) of common carbohydrates. The results proved PF-06305591 that TZQ could decrease the GI of sucrose, maltose and starch. For maltose, 6 tablets of TZQ were the best dose, and the activity of maltase was inhibited. Based on these results, this study aimed to investigate the mechanisms of PF-06305591 TZQ action on maltase by measuring maltase inhibitory activity, conducting Rabbit polyclonal to EPHA4 kinetics assays and determining the secondary structures of maltase via circular dichroism (CD). Acarbose was used as a control drug. This study will provide a scientific basis for TZQ treatment of diabetes and useful in new drug development. Methods Materials Maltase (EC: 22.214.171.124) was purchased from Shanghai Yuanye Bio-Technology Co., Ltd. (Shanghai China). Acarbose was obtained from the National Institutes for Food and Drug Control (Beijing, China). Maltose was purchased from TS Corporation (Seoul, Korea). TZQ extract was provided by Shandong Buchang Shenzhou Pharmaceutical Co., Ltd. The content of TZQ was determined by the chemical marker component of hypericin (10?mg/g TZQ). All other reagents and solvents were of analytical reagent grade, and ultrapure water was used throughout the experiment. Excess concentration of the substrate, optimum concentration of the enzyme and optimum reaction time Ten microlitres of maltase (5?mg/mL) was reacted with maltase substrates at different concentrations (0.0125, 0.025, 0.05, 0.1, 0.175, 0.25, 0.40, and 0.55?mol/L) at 37?C for 10?min and cooled.