Inhibitory effects of chemical inhibitors Ko143 and MK571 on demethoxycurcumin-<i>O</i>-glucuronidation activity.

(A) Inhibitory effects of chemical inhibitors on glucuronidation activity by UGT1A1; (B) Inhibitory effects of chemical inhibitors on glucuronidation activity by HeLa1A1 cell lysate. The concentration of demethoxycurcumin was 4 μM. All experiments were performed in triplicate (n = 3). Data were expressed as mean ± SD (n = 3). * p p p # p ## p ### p < 0.001 compared with that of Ko143 (5 μM) or MK571 (5 μM), respectively.</p

Effects of Ko143 on the formation and efflux excretion of demethoxycurcumin-<i>O</i>-glucuronides in HeLa1A1 cells.

(A) Effects of Ko143 (5 and 20 μM) on the accumulation of demethoxycurcumin-O-glucuronides (G1) in extracellular medium; (B) Effects of Ko143 (5 and 20 μM) on demethoxycurcumin-O-glucuronides (G2) excretion in extracellular solutions; (C) Effects of Ko143 (5 and 20 μM) on the efflux excretion rates of G1 and G2; (D) Effects of Ko143 (5 and 20 μM) on the intracellular G1 and G2 levels; (E) Effects of Ko143 (5 and 20 μM) on the fraction metabolized (fmet) of demethoxycurcumin; All experiments were performed in triplicate (n = 3). Data were presented as mean ± SD (n = 3). *, # p, ## p, ### p<0.001 compared with that of control group of G1 or G2, respectively.</p

UGT1A1 catalyzed the demethoxycurcumin-<i>O</i>-glucuronidation, and expression of UGT1A1 and efflux transporters in HeLa1A1 cells.

(A) Kinetic profiles for demethoxycurcumin-O-glucuronides (G1 and G2) (0.5–40 μM) by UGT1A1; (B) Kinetic profiles for demethoxycurcumin-O-glucuronides (G1 and G2) (0.5–40 μM) by HeLa1A1 cell lysate; In each panel, the insert figure showed the corresponding Eadie-Hofstee plot. (C) Protein expression of UGT1A1, BCRP and four MRP family transporters in HeLa and HeLa1A1 cells. All experiments were performed in triplicate (n = 3). Data were expressed as mean ± SD (n = 3).</p

Metabolism and disposition of corylifol A from <i>Psoralea corylifolia</i>: metabolite mapping, isozyme contribution, species differences and identification of efflux transporters for corylifol A-<i>O</i>-glucuronide in HeLa1A1 cells

Corylifol A (CA), a phenolic compound from Psoralea corylifolia, possessed several biological properties but poor bioavailability. Here we aimed to investigate the roles of cytochromes P450s (CYPs), UDP-glucuronosyltransferases (UGTs) and efflux transporters in metabolism and disposition of CA.Metabolism of CA was evaluated in HLM, expressed CYPs and UGTs. Chemical inhibitors and shRNA-mediated gene silencing of multidrug resistance-associated proteins (MRPs) and breast cancer resistance protein (BCRP) were performed to assess the roles of transporters in CA disposition.Three oxidated metabolites (M1–M3) and two glucuronides (M4–M5) were detected. The intrinsic clearances (CLint) values of M1 and M4 in HLM were 48.10 and 184.03 μL/min/mg, respectively. Additionally, CYP1A1, 2C8 and 2C19 were identified as main contributors with CLint values of 13.01–49.36 μL/min/mg, while UGT1A1, 1A7, 1A8 and 1A9 were with CLint values ranging from 85.01 to 284.07 μL/min/mg. Furthermore, activity correlation analysis proved CYP2C8, UGT1A1 and 1A9 were the main active hepatic isozymes. Besides, rats and monkeys were appropriate model animals. Moreover, dipyridamole and MK571 both could significantly inhibit M4 efflux. Gene silencing results also indicated MRP4 and BCRP were major contributors in HeLa1A1 cells.Taken together, CYPs, UGTs, MRP4 and BCRP were important determinants of CA pharmacokinetics. Corylifol A (CA), a phenolic compound from Psoralea corylifolia, possessed several biological properties but poor bioavailability. Here we aimed to investigate the roles of cytochromes P450s (CYPs), UDP-glucuronosyltransferases (UGTs) and efflux transporters in metabolism and disposition of CA. Metabolism of CA was evaluated in HLM, expressed CYPs and UGTs. Chemical inhibitors and shRNA-mediated gene silencing of multidrug resistance-associated proteins (MRPs) and breast cancer resistance protein (BCRP) were performed to assess the roles of transporters in CA disposition. Three oxidated metabolites (M1–M3) and two glucuronides (M4–M5) were detected. The intrinsic clearances (CLint) values of M1 and M4 in HLM were 48.10 and 184.03 μL/min/mg, respectively. Additionally, CYP1A1, 2C8 and 2C19 were identified as main contributors with CLint values of 13.01–49.36 μL/min/mg, while UGT1A1, 1A7, 1A8 and 1A9 were with CLint values ranging from 85.01 to 284.07 μL/min/mg. Furthermore, activity correlation analysis proved CYP2C8, UGT1A1 and 1A9 were the main active hepatic isozymes. Besides, rats and monkeys were appropriate model animals. Moreover, dipyridamole and MK571 both could significantly inhibit M4 efflux. Gene silencing results also indicated MRP4 and BCRP were major contributors in HeLa1A1 cells. Taken together, CYPs, UGTs, MRP4 and BCRP were important determinants of CA pharmacokinetics.</p

Effects of knock-down of MRP1 transporter on demethoxycurcumin-<i>O</i>-glucuronides excretion and cellular glucuronidation in HeLa1A1 cells.

(A) Effects of accumlated demethoxycurcumin-O-glucuronides (G1) in extracellular medium; (B) Effects of excreted demethoxycurcumin-O-glucuronides (G2) in extracellular medium; (C) Effects of the excretion rates of G1 and G2; (D) Effects of the intracellular G1 and G2 levels; (E) Effects of fmet values of demethoxycurcumin; (F) Effects of gene silencing on the protein level of MRP1. The concentration of demethoxycurcumin was 4 μM. All experiments were performed in triplicate (n = 3). Data were expressed as mean ± SD (n = 3). *, # p , ## p , ### p < 0.001 compared with that of control group of G1 or G2, respectively.</p

MRP3 silencing led to reduce excretion rates and increase intracellular demethoxycurcumin-<i>O</i>-glucuronides in HeLa1A1 cells.

(A) MRP3 transporter silencing inhibited the accumlated demethoxycurcumin-O-glucuronides (G1) in extracellular medium; (B) MRP3 transporter silencing decreased the excreted demethoxycurcumin-O-glucuronides (G2) in extracellular solution; (C) MRP3 silencing significantly reduced the excretion rates of G1 and G2; (D) MRP3 silencing resulted in evaluation of intracellular G1 and G2 levels; (E) MRP3 silencing decreased total cellular glucuronidation (G1 ang G2) of demethoxycurcumin; (F) Effects of gene silencing on the protein level of MRP3. The concentration of demethoxycurcumin was 4 μM. All experiments were performed in triplicate (n = 3). Data were presented as mean ± SD. *, # p , ## p , ### p < 0.001 compared with that of control group of G1 or G2, respectively.</p

Effects of shRNA-mediated MRP4 transporter knock-down on demethoxycurcumin-<i>O</i>-glucuronides excretion and cellular glucuronidation in HeLa1A1 cells.

(A) MRP4 knock-down decreased the excreted demethoxycurcumin-O-glucuronides (G1); (B) MRP4 knock-down reduced the accumlated demethoxycurcumin-O-glucuronides (G2) in extracellular medium; (C) MRP4 knock-down inhibited the excretion rates of G1 and G2; (D) The intracellular G1 and G2 levels were evaluated when MRP4 transporter was knock-down; (E) MRP4 knock-down resulted in decrease of overall cellular glucuronidation (G1 and G2) of demethoxycurcumin; (F) Effects of gene silencing on the protein level of MRP4. The concentration of demethoxycurcumin was 4 μM. All experiments were performed in triplicate (n = 3). Data were expressed as mean ± SD (n = 3). *, # p , ## p , ### p < 0.001 compared with that of control group of G1 or G2, respectively.</p

Data_Sheet_1_Characterization of volatile organic compounds with anti-atherosclerosis effects in Allium macrostemon Bge. and Allium chinense G. Don by head space solid phase microextraction coupled with gas chromatography tandem mass spectrometry.docx

IntroductionAllium macrostemon Bge. (AMB) and Allium chinense G. Don (ACGD) are both edible Allium vegetables and named officinal Xiebai (or Allii Macrostemonis Bulbus) in East Asia. Their medicinal qualities involve in lipid lowering and anti-atherosclerosis effects. And steroidal saponins, nitrogenous compounds and sulfur compounds are like the beneficial components responsible for medicinal functions. Sulfur compounds are the recognized main components both in the volatile oils of AMB and ACGD. Besides, few researches were reported about their holistic chemical profiles of volatile organic compounds (VOCs) and pharmacodynamic effects.MethodsIn this study, we first investigated the lipid-lowering and anti-atherosclerotic effects of volatile oils derived from AMB and ACGD in ApoE–/– mice with high fat and high cholesterol diets.ResultsThe results showed the volatile oils of AMB and ACGD both could markedly reduce serum levels of TG, TC, and LDL-C (p 0.05). Pathological results displayed they both could obviously improve the morphology of cardiomyocytes and the degree of myocardial fibrosis in model mice. Meanwhile, oil red O staining results also proved they could apparently decrease the lesion areas of plaques in the aortic intima (p DiscussionTaken together, this study was the first analysis of holistic chemical profiles and anti-atherosclerosis effects of AMB and ACGD volatile oils, and would benefit the understanding of effective components in AMB and ACGD.</p

DataSheet1_Population Pharmacokinetics of Mycophenolic Acid in Renal Transplant Patients: A Comparison of the Early and Stable Posttransplant Stages.docx

Mycophenolic acid (MPA) is an antimetabolic immunosuppressive drug widely used in solid organ transplantation and autoimmune diseases. Pharmacokinetics (PK) of MPA demonstrates high inter- and intra-variability. The aim of this study was to compare the population PK properties of MPA in adult renal transplant patients in the early and stable post-transplant stages and to simulate an optimal dosing regimen for patients at different stages. A total of 51 patients in the early post-transplant period (1 week after surgery) and 48 patients in the stable state (5.5–10 years after surgery) were included in the study. In the two-compartment population PK model, CL/F (23.36 L/h vs. 10.25 L/h) and V/F (78.07 vs. 16.24 L) were significantly different between the two stages. The dose-adjusted area under the concentration time curve (AUCss,12h/dose) for patients in the early stage were significantly lower than those for patients in the stable state (40.83 ± 22.26 mg h/L vs. 77.86 ± 21.34 mg h/L; p ss,12h of 30–60 mg h/L. In addition, limited sampling strategies showed that two 4-point models (C0-C1-C2-C4 and C1-C2-C3-C6) performed well in predicting MPA exposure by both Bayesian estimate and regression equation and could be applied in clinical practice to assist therapeutic drug monitoring of MPA.</p