Yanfen Li

Systemic Exposure to and Disposition of Catechols Derived from Salvia miltiorrhiza Roots (Danshen) after Intravenous Dosing DanHong Injection in Human Subjects, Rats, and Dogs

DanHong injection is a Danshen (Salvia miltiorrhiza roots)-based injectable solution for treatment of coronary artery disease and ischemic stroke. Danshen catechols are believed to be responsible for the injection’s therapeutic effects. This study aimed to characterize systemic exposure to and elimination of Danshen catechols in human subjects, rats, and dogs receiving intravenous DanHong injection. A total of 28 catechols were detected, with content levels of 0.002–7.066 mM in the injection, and the major compounds included tanshinol, protocatechuic aldehyde, salvianolic acid B, rosmarinic acid, salvianolic acids A and D, and lithospermic acid with their daily doses ≥10 μmol/subject. After dosing, tanshinol, salvianolic acid D, and lithospermic acid exhibited considerable exposure in human subjects and rats. However, only tanshinol had considerable exposure in dogs. The considerable exposure to tanshinol was due to its having the highest dose, whereas that to salvianolic acid D and lithospermic acid was due to their relatively long elimination half-lives in the human subjects and rats. Protocatechuic aldehyde and rosmarinic acid circulated in the bloodstream predominantly as metabolites; salvianolic acids A and B exhibited low plasma levels with their human plasma metabolites little or not detected. Tanshinol and salvianolic acid D were eliminated mainly via renal excretion. Elimination of other catechols involved hepatobiliary and/or renal excretion of their metabolites. Methylation was found to be the primary metabolism for most Danshen catechols and showed intercompound and interspecies differences in rate and degree in vitro. The information gained here is relevant to pharmacological and toxicological research on DanHong injection.

Stability of immobilized R. oryzae in repetitive batch productions of l(+)-lactic acid : effect of inorganic salts

Rhizopus oryzae was immobilized in polyurethane foam cubes by a natural attachment method. The effect of inorganic salts on the stability of the immobilized mycelium in repetitive batch productions of L(+)-lactic acid was studied. The amount of the inorganic salts necessary to maintain the activity of the immobilized R. oryzae in the repetitive batch fermentations strongly depended upon the initial glucose concentrations. For example, the amount of the inorganic salts should be doubled if the initial glucose concentration was twice increased. The minimum amounts of the inorganic salts were therefore determined for effective lactic acid productions in the repetitive batch fermentations with the immobilized R. oryzae.

Pharmacokinetics and disposition of monoterpene glycosides derived from Paeonia lactiflora roots (Chishao) after intravenous dosing of antiseptic XueBiJing injection in human subjects and rats

Aim:Monoterpene glycosides derived from Paeonia lactiflora roots (Chishao) are believed to be pharmacologically important for the antiseptic herbal injection XueBiJing. This study was designed to characterize the pharmacokinetics and disposition of monoterpene glycosides.Methods:Systemic exposure to Chishao monoterpene glycosides was assessed in human subjects receiving an intravenous infusion and multiple infusions of XueBiJing injection, followed by assessment of the pharmacokinetics of the major circulating compounds. Supportive rat studies were also performed. Membrane permeability and plasma-protein binding were assessed in vitro.Results:A total of 18 monoterpene glycosides were detected in XueBiJing injection (content levels, 0.001–2.47 mmol/L), and paeoniflorin accounted for 85.5% of the total dose of monoterpene glycosides detected. In human subjects, unchanged paeoniflorin exhibited considerable levels of systemic exposure with elimination half-lives of 1.2–1.3 h; no significant metabolite was detected. Oxypaeoniflorin and albiflorin exhibited low exposure levels, and the remaining minor monoterpene glycosides were negligible or undetected. Glomerular-filtration-based renal excretion was the major elimination pathway of paeoniflorin, which was poorly bound to plasma protein. In rats, the systemic exposure level of paeoniflorin increased proportionally as the dose was increased. Rat lung, heart, and liver exposure levels of paeoniflorin were lower than the plasma level, with the exception of the kidney level, which was 4.3-fold greater than the plasma level; brain penetration was limited by the poor membrane permeability.Conclusion:Due to its significant systemic exposure and appropriate pharmacokinetic profile, as well as previously reported antiseptic properties, paeoniflorin is a promising XueBiJing constituent of therapeutic importance.

Pharmacokinetics of catechols in human subjects intravenously receiving XueBiJing injection, an emerging antiseptic herbal medicine

XueBiJing injection, prepared from a five-herb combination, is extensively used as add-on therapy in routine sepsis care in China. Catechols, derived from the component herb Salvia miltiorrhiza roots (Danshen), are probably important because of their reported antiseptic properties. This study was designed to characterize pharmacokinetics of major circulating Danshen catechols in human subjects intravenously receiving the injection at the label doses. A total of 17 Danshen catechols were detected in XueBiJing injection (content level, 0.1-139.3 μmol/L). After dosing, tanshinol and salvianolic acid B exhibited relatively high levels of systemic exposure with mean elimination half-lives of 0.38 and 0.29 h, respectively. The total plasma clearance and apparent volume of distribution at steady state of tanshinol were 1.07 L/h/kg and 0.40 L/kg, respectively, whereas those of salvianolic acid B were 0.43 L/h/kg and 0.13 L/kg, respectively. Protocatechuic acid and five other catechols were also detected in plasma but at low exposure levels. Although protocatechuic aldehyde had the highest content level in the injection, like the remaining eight catechols, it was undetected in plasma. Protocatechuic aldehyde was extensively converted into protocatechuic acid and other metabolites. The information gained here facilitates understanding the roles of Danshen catechols in therapeutic actions of XueBiJing injection.

Effect of danhong injection on the mobilisation of endothelial progenitor cells to vascular repair after percutaneous coronary intervention: a randomised controlled trial

BACKGROUNDnEndothelial progenitor cells (EPCs) derived from bone marrow are associated with the repair of vascular injury by accelerating re-endothelisation, and represent a potential therapeutic target for ischaemic disease. Danhong injection (DHI), a Chinese Materia Medica standardised product extracted from Radix Salvia miltiorrhiza and Flos Carthamus tinctorius L, is effective in the treatment of ischaemic heart disease. We assessed the effect of DHI on the mobilisation of EPCs and vascular repair mediated by EPCs after percutaneous coronary intervention (PCI).nnnMETHODSnIn a prospective, randomised clinical trial (ChiCTR-ONRC-14004100), patients with coronary heart disease and chest pain (n=54) who were referred to coronary angiography and underwent PCI were enrolled; 12 dropped out because of incomplete blood sample collection. The remaining 42 patients (age 35-75 years) were randomly assigned by random number table method to either a control group (14 men, seven women) that received standard medical treatment or a DHI group (14 men, seven women) that received standard medical treatment combined with DHI (40 mL per day) for 7 days at the affiliated cardiovascular hospital of Qingdao University. The primary outcomes were endothelial lesion and endothelial regeneration. Endothelial lesion was assessed by enumeration of CD45- CD146+ KDR+ circulating endothelial cells (CECs) and inflammatory factors (interleukin 6, C-reactive protein [CRP], tumour necrosis factor α [TNFα]). Endothelial regeneration was evaluated by measurement of CD45dim CD34+ KDR+ EPCs, as determined by flow cytometry. Measurements were taken before PCI, 1 day after PCI, and 7 days after treatment by two investigators masked to treatment assignment. All healthy participants and those with coronary heart disease gave informed consent to participate in the study. All protocols were approved by each of the local ethics committees.nnnFINDINGSnBaseline cytokines, CECs, and EPCs did not differ between DHI and control groups. PCI increased interleukin 6 (p=0·004 in control group; p=0·045 in DHI group), TNF α (p=0·001 in control group; p=0·16 in DHI group), and CECs concentrations (p=0·008 in control group; p=0·005 in DHI group), whereas EPCs decreased (p=0·002 in control group; p=0·001 in DHI group). Compared with the control group, the mean expression levels of cytokines were significantly reduced (interleukin 6 14·98 pg/mL [SD 13·51] vs 7·25 pg/mL [8·03], p=0·001; TNFα 594·17 pg/mL [368·34] vs 388·48 pg/mL [210·24], p<0·0001; CRP 38·07 ng/mL [18·93] vs 22·42 ng/mL [5·94], p=0·04), and CECs were significantly reduced (0·1228% of PBMC [0·0669] vs 0·0564% of PBMC [0·0445], p<0·0001). EPC population significantly increased (0·0962% of PBMC [0·044] vs 0·2569 of PBMC [0·1215], p<0·0001) in DHI group after treatment.nnnINTERPRETATIONnThese initial results suggest that DHI is effective in improving endothelial repair and protecting the endothelial lesion by mobilising EPCs, by inhibiting the inflammatory response, and by inhibiting of shedding of endothelial cells after PCI in patients with coronary heart disease.nnnFUNDINGnChina National Basic Research Program (973 programme, number 2012CB518404).

Nuciferine and paeoniflorin can be quality markers of Tangzhiqing tablet, a Chinese traditional patent medicine, based on the qualitative, quantitative and dose-exposure-response analysis

BACKGROUNDnDiabetes is a chronic disease associated with significant morbidity and mortality. Tangzhiqing tablet (TZQ), a Chinese traditional patent medicine, has been in phase 2 clinical trial for the treatment of diabetes mellitus. However, the current quality evaluation of TZQ still remains rather obscure.nnnPURPOSEnThe promising quality markers (Q-markers) of TZQ will be sought for its quality assessment and process control based on the qualitative, quantitative and dose-exposure-response analysis.nnnMETHODSnThe fingerprint analysis of TZQ was carried out through ultra high performance liquid chromatography- quadrupole time-of-flight/ mass spectrometry (UPLC-Q-TOF/MS) assay. Multicomponent quantitative analysis was implemented to the main ingredients of nuciferine, paeoniflorin, salvianolic acid B, hyperoside and rutin in TZQ by means of LC analysis. The dose-exposure-response relationship of TZQ was revealed by a placebo-controlled, 5-way crossover study in healthy Chinese subjects. The potential Q-markers in plasma were determined by a liquid chromatography combined with tandem mass spectrometry (LC-MS/MS) method. The therapeutic effect of TZQ was expressed as the glucose-lowering profile. The exposure-response relevance was developed between the concentration of Q-markers and the glucose-lowering effect of TZQ.nnnRESULTSn46 compounds were identified or tentatively characterized from TZQ. The contents of paeoniflorin, nuciferine, salvianolic acid B, hyperoside and rutin in TZQ were 6.40, 1.75, 1.70, 0.004, and 0.006u202fmg, respectively. However, salvianolic acid B, hyperoside and rutin could hardly be detected in human plasma under the current LC-MS/MS condition. The exposures of nuciferine and paeoniflorin (AUC0-3, Cmax) were dose-proportionality in human at the studied dosage ranges. The glucose-lowering effect appeared to increase proportionally with increasing TZQ dose in healthy volunteers. A clockwise hysteresis was displayed between the exposure of nuciferine and paeoniflorin and the glucose-lowering effect of TZQ.nnnCONCLUSIONnNuciferine and paeoniflorin were identified as the promising Q-markers of TZQ based on the fingerprint qualitative analysis, multicomponent quantitative analysis and dose-exposure-response analysis. The two Q-markers are meaningful to ensure the quality assessment and process control of TZQ.

Identify super quality markers from prototype-based pharmacokinetic markers of Tangzhiqing tablet (TZQ) based on in vitro dissolution/ permeation and in vivo absorption correlations

BACKGROUNDnA quality marker (Q-marker) is defined as an inherent chemical compound that is used for the quality control of a drug. Its biological activities are closely related to safety and therapeutic effects. Generally, a multiple-component herbal medicine may have many Q-markers. We therefore proposed a concept of super Q-marker satisfying both the criterion of Q-markers and PK-markers to be used in more effective quality control of herbal medicine.nnnPURPOSEnThe first aim was to find suitable prototype-based PK-markers from Tangzhiqing tablets (TZQ), a Chinese patent medicine. Then super Q-markers were expected to be identified from the prototype-based PK-markers based on an in vitro-in vivo correlation study.nnnMETHODSnPotentially eligible prototype-based PK-markers were identified in a single- and multiple-dose pharmacokinetic study on TZQ in 30 healthy volunteers. The in vitro dissolution and permeation profiles of the prototype-based PK-markers of TZQ were evaluated by the physiologically-based drug dissolution/absorption simulating system (DDASS). An in vitro-in vivo correlation analysis was conducted between the dissolution/permeation behaviors in DDASS and the actual absorption profiles in human to test the transferability and traceability of the promising super Q-markers for TZQ.nnnRESULTSnIn human, plasma paeoniflorin and nuciferine as prototype-based PK-markers exhibited the appropriate pharmacokinetic properties, including dose-dependent systemic exposure (AUC, Cmax) and a proper elimination half-life (1∼3h). In DDASS, it was predicted that paeoniflorin and nuciferine are highly permeable but the absorption rates are primarily limited by the dissolution rates. Moreover, the established in vitro-in vivo correlations of paeoniflorin and nuciferine were in support of the super Q-markers features.nnnCONCLUSIONnPaeoniflorin and nuciferine are identified as the super Q-markers from the prototype-based PK-markers of TZQ based on findings from a combination of in vitro, in vivo, and in vitro-in vivo correlation studies. This method is practical for optimal identification of qualified Q-markers, thus helping improve the quality control of herbal medicines.

Metabolomics-Based Clinical Efficacy and Effect on the Endogenous Metabolites of Tangzhiqing Tablet, a Chinese Patent Medicine for Type 2 Diabetes Mellitus with Hypertriglyceridemia

Tangzhiqing tablet (TZQ) is derived from Tangzhiqing formula, which has been used to regulate glucose and lipid metabolism in China for hundreds of years. However, as a new Chinese patent medicine, its clinical indication is not clear. To explore the clinical indication and effect on the patients with type 2 diabetes mellitus (T2DM), a pilot clinical trial and metabolomics study were carried out. In the clinical study, T2DM patients were divided into three groups and treated with TZQ, placebo, or acarbose for 12 weeks, respectively. The metabolomic study based on UPLC Q-TOF MS was performed including patients with hypertriglyceridemia in TZQ and placebo groups and healthy volunteers. The clinical results showed that TZQ could reduce glycosylated hemoglobin (HbA1c) and fasting insulin. For patients with hypertriglyceridemia in TZQ group, the levels of HbA1c all decreased and were correlated with the baseline level of triglyceride. Metabonomics data showed a significant difference between patients and healthy volunteers, and 17 biomarkers were identified. After 12-week treatment with TZQ, 11 biomarkers decreased significantly (p<0.05), suggesting that TZQ could improve the metabolomic abnormalities in these participants. In conclusion, the clinical indication of TZQ was T2DM with hypertriglyceridemia, and its target was related to glycerophospholipid metabolism.

Pharmacokinetics-Based Identification of Potential Therapeutic Phthalides from XueBiJing, a Chinese Herbal Injection Used in Sepsis Management

XueBiJing, an injectable five-herb preparation, has been incorporated into routine sepsis care in China. Phthalides, originating from XueBiJing’s component herbs Ligusticum chuanxiong rhizomes and Angelica sinensis roots, are believed to contribute to its therapeutic effects due to their presence in the preparation and antisepsis-related properties. This investigation aimed to identify potential therapeutic phthalides that are bioavailable to act on XueBiJing’s therapeutic targets and that could serve as pharmacokinetic markers to supplement classic biomarkers for sepsis care. Among 10 phthalides detected in XueBiJing, senkyunolides I and G were the major circulating phthalides in human subjects, but their different pharmacokinetics might influence their contribution to XueBiJing’s therapeutic action. Senkyunolide I exhibited a large distribution volume (1.32 l/kg) and was moderately bound in plasma (54% unbound), whereas senkyunolide G exhibited a small distribution volume (0.10 l/kg) and was extensively bound in plasma (3% unbound). Clearance of senkyunolide I from the systemic circulation was governed by UGT2B15-mediated hepatic glucuronidation; the resulting electrophilic glucuronides were conjugated with glutathione in the liver. Senkyunolide G was selectively bound to albumin (99%) in human plasma. To our knowledge, the human pharmacokinetic data of XueBiJing’s phthalides are reported here for the first time. Based on this investigation and such investigations of the other component herbs, follow-up pharmacodynamic assessments of bioavailable herbal compounds are planned to elucidate XueBiJings chemical basis responsible for its therapeutic action. Senkyunolides I and G, having the preceding disposition characteristics that could be detectably altered by septic pathophysiology, could serve as pharmacokinetic markers for sepsis care.

Human pharmacokinetics of ginkgo terpene lactones and impact of carboxylation in blood on their platelet-activating factor antagonistic activity

Terpene lactones are a class of bioactive constituents of standardized preparations of Ginkgo biloba leaf extract, extensively used as add-on therapies in patients with ischemic cardiovascular and cerebrovascular diseases. This investigation evaluated human pharmacokinetics of ginkgo terpene lactones and impact of their carboxylation in blood. Human subjects received oral YinXing-TongZhi tablet or intravenous ShuXueNing, two standardized ginkgo preparations. Their plasma protein-binding and platelet-activating factor antagonistic activity were assessed in vitro. Their carboxylation was assessed in phosphate-buffered saline (pH 7.4) and in human plasma. After dosing YinXing-TongZhi tablet, ginkgolides A and B and bilobalide exhibited significantly higher systemic exposure levels than ginkgolides C and J; after dosing ShuXueNing, ginkgolides A, B, C, and J exhibited high exposure levels. The compounds’ unbound fractions in plasma were 45–92%. Apparent oral bioavailability of ginkgolides A and B was mostly >100%, while that of ginkgolides C and J was 6–15%. Bilobalide’s bioavailability was probably high but lower than that of ginkgolides A/B. Terminal half-lives of ginkgolides A, B, and C (4–7u2009h) after dosing ShuXueNing were shorter than their respective values (6–13u2009h) after dosing YinXing-TongZhi tablet. Half-life of bilobalide after dosing the tablet was around 5u2009h. Terpene lactones were roughly evenly distributed in various body fluids and tissues; glomerular-filtration-based renal excretion was the predominant elimination route for the ginkgolides and a major route for bilobalide. Terpene lactones circulated as trilactones and monocarboxylates. Carboxylation reduced platelet-activating factor antagonistic activity of ginkgolides A, B, and C. Ginkgolide J, bilobalide, and ginkgo flavonoids exhibited no such bioactivity. Collectively, differences in terpene lactones’ exposure between the two preparations and influence of their carboxylation in blood should be considered in investigating the relative contributions of terpene lactones to ginkgo preparations’ therapeutic effects. The results here will inform rational clinical use of ginkgo preparations.