Alstonia scholaris(L.)R.Br.,an evergreen tropical plant rich in indole alkaloids with significant physiological activity,is traditionally used to treat respiratory diseases in China.This study was conducted to establi...Alstonia scholaris(L.)R.Br.,an evergreen tropical plant rich in indole alkaloids with significant physiological activity,is traditionally used to treat respiratory diseases in China.This study was conducted to establish the toxicity profile of the alkaloid extract(TA)of A.scholaris leaves in non-rodents.After oral administration of a single dose(4 g/kg.bw),a num-ber of transient symptoms,such as unsteady gait,drooling,emesis,and reddening of peri-oral mucosa,were observed,but no treatment-related mortality.A sub-chronic toxicity study with a range of doses of TA(20,60 and 120 mg/kg.bw)was conducted for a 13-week treatment period,followed by 4-week recovery observation.Except for emesis and drooling in majority of animals in 120 mg/kg.bw treatment group,no clinical changes were observed in TA-treated animals.Data from electrocardiography,bone marrow,urine,fecal,hematology and clinical chemistry analyses were comparable between TA-treated and control animals.No significant differences in the relative organ weights and histopathological characteristics were evident between the TA-treated and control groups.Accordingly,the non-observed-adverse-effect-level(NOAEL)of TA was established as 120 mg/kg.bw.Our results add further knowledge to the safety database for indole alkaloid extracts from A.scholaris with potential utility as novel drug candidates.展开更多
Shenfu Injection(SFI) is a well-defined Chinese herbal formulation that is obtained from red ginseng and processed aconite root. The main active constituents in SFI are ginsenosides and aconitum alkaloids. In this wor...Shenfu Injection(SFI) is a well-defined Chinese herbal formulation that is obtained from red ginseng and processed aconite root. The main active constituents in SFI are ginsenosides and aconitum alkaloids. In this work, ginsenosides(ginsenoside Rg1, ginsenoside Rb1 and ginsenoside Rc) and aconitum alkaloids(benzoylmesaconine and fuziline) were used as the index components to explore the pharmacokinetic behavior of SFI. A selective and sensitive HPLC–MS/MS method was developed for the quantification of ginsenosides and aconitum alkaloids in dog plasma and was used to characterize the pharmacokinetics of the five index components after intravenous drip of three different dosages of SFI in beagle dogs. The pharmacokinetic properties of the index components were linear over the dose range of2–8 m L/kg.展开更多
This paper describes the development and validation of a liquid chromatography–mass spectrometric assay for propafenone and its application to a pharmacokinetic study of propafenone administered as a new propafenone ...This paper describes the development and validation of a liquid chromatography–mass spectrometric assay for propafenone and its application to a pharmacokinetic study of propafenone administered as a new propafenone hydrochloride sustained-release capsule(SR-test), as an instant-release tablet(IR-reference) and as the market leader sustained-release capsule(Rythmol, SR-reference) in male beagle dogs(n ? 8). In Study A comparing SR-test with IR-reference in a crossover design T_(max) and t_(1/2) of propafenone for SR-test were significantly higher than those for IR-reference while C_(max) and AUC were lower demonstrating the sustained release properties of the new formulation. In Study B comparing SRtest with SR-reference the observed C_(max) and AUC of propafenone for SR-test(124.57140.0 ng/m L and612.07699.2 ng·h/m L, respectively) were higher than for SR-reference(78.52772.92 ng/m L and423.67431.6 ng·h/m L, respectively) although the differences were not significant. Overall, the new formulation has as good if not better sustained release characteristics to the market leader formulation.展开更多
Retinal injury after blunt ocular trauma may directly affect prognosis and lead to vision loss.To investigate the pathological changes and molecular mechanisms involved in retinal injury after blunt ocular trauma,we e...Retinal injury after blunt ocular trauma may directly affect prognosis and lead to vision loss.To investigate the pathological changes and molecular mechanisms involved in retinal injury after blunt ocular trauma,we established a weight drop injury model of blunt ocular trauma in male Beagle dogs.Hematoxylin-eosin staining,immunofluorescence staining,western blotting,and TUNEL assays were performed to investigate retinal injury within 14 days after blunt ocular trauma.Compared with the control group,the thicknesses of the inner and outer nuclear layers,as well as the number of retinal ganglion cells,gradually decreased within 14 days after injury.The number of bipolar cells in the inner nuclear layer began to decrease 1 day after injury,while the numbers of cholinergic and amacrine cells in the inner nuclear layer did not decrease until 7 days after injury.Moreover,retinal cell necroptosis increased with time after injury;it progressed from the ganglion cell layer to the outer nuclear layer.Visual electrophysiological findings indicated that visual impairment began on the first day after injury and worsened over time.Additionally,blunt ocular trauma induced nerve regeneration and Müller glial hyperplasia;it also resulted in the recruitment of microglia to the retina and polarization of those microglia to the M1 phenotype.These findings suggest that necroptosis plays an important role in exacerbating retinal injury after blunt ocular trauma via gliosis and neuroinflammation.Such a role has important implications for the development of therapeutic strategies.展开更多
Objective To develop and validate a simple, rapid, sensitive, and reproducible HPLC method for determination of hyperoside in plasma of dogs and for the subsequent pharmacokinetic (PK) study. Methods An accurate and r...Objective To develop and validate a simple, rapid, sensitive, and reproducible HPLC method for determination of hyperoside in plasma of dogs and for the subsequent pharmacokinetic (PK) study. Methods An accurate and reproducible HPLC-UV method was developed and validated for the determination of hyperoside in plasma of dogs, using kaempferol as internal standard. The plasma samples of dogs following ig administration of hyperoside were analyzed for the detection of quercetin after enzymatic hydrolysis treatment with combined β-glucuronidase and sulphatase. The analytes were separated on a Diamonsil C 18 column (250 mm × 4.6 mm, 5 μm). The mobile phase consisted of methanol-buffer solution (0.1 mol/L NH 4 Ac + 0.3 mmol/L EDTA-Na 2 )-acetic acid (60:40:1) and was delivered at a flow rate of 1 mL/min. The UV detector was set at 370 nm and the column temperature was maintained at 35 ℃. The sample injection volume was 20 μL. Data were collected and analyzed using the ANASTAR software. PK parameters were calculated with DAS software (2.0). Results Linear relationships were validated over the range of 0.01-1 μg/mL for hyperoside (r = 0.9997). The intra- and inter-day precision values for all samples were within 10.0%, and the accuracies of intra- and inter-day assays were within the range of 92.4%-102.4%. The validated method was successfully used to determine the hyperoside concentration in plasma of dogs for up to 12 h, after a single ig administration (25 mg/kg). The mean PK parameters for male and female dogs were as follows: C max (0.18 ± 0.05) and (0.16 ± 0.05) μg/mL, AUC 0-∞ (0.79 ± 0.34) and (0.86 ± 0.27) μg/(mL·h), t 1/2(ka) (0.89 ± 0.41) and (0.88 ± 0.28) h, respectively. Statistical analysis on the PK of hyperoside in male and female groups showed that sex had no significant impact on the PK of hyperoside (P > 0.05). Conclusion The method is able and sufficient to be used in drug PK studies of hyperoside.展开更多
Objective To develop an LC-MS/MS method for determining the concentration of wogonin in dog plasma and investigate the pharmacokinetics and bioavailability by different administrations of wogonin in Beagle's dogs....Objective To develop an LC-MS/MS method for determining the concentration of wogonin in dog plasma and investigate the pharmacokinetics and bioavailability by different administrations of wogonin in Beagle's dogs.Methods LC-MS/MS was employed in determining the concentration of wogonin with the selected ion monitoring model after liquid-liquid extraction with ethyl acetate of dog plasma samples.The lower limit of quantification was 0.105 μg/L.Target ions were at m/z 285.0→270.0 for wogonin and 373.3→305.3 for finasteride.In a randomized,self-control,and cross-over study,six male Beagle's dogs were treated with different administration methods in three test periods.Pharmacokinetic parameters were calculated with DAS software(Ver.2.0).Results The calibration curve was linear in the range of 0.105-107.36 μg/L for wogonin in dog plasma samples.The main pharmacokinetic parameters of ig administration(native drug of 15 mg/kg and solution preparation of 5 mg/kg) and iv route were as follows:Cmax(2.5 ± 1.1),(7.9 ± 3.3),and(6838.7 ± 1322.1) μg/L,tmax(0.7 ± 0.3) and(0.3 ± 0.2) h for the both former,AUC0-t(7.1 ± 2.0),(21.0 ± 3.2),and(629.7 ± 111.8) μg.h/L.The absolute bioavailability of native and solution of wogonin were(0.59 ± 0.35)% and(3.65 ± 2.00)%,respectively.Conclusion The validated method is convenient,sensitive,and specific,and the improvement of wogonin solubility could remarkably increase the absolute bioavailability.展开更多
Objective:To simultaneously investigate the pharmacokinetics of gentiopicroside,sweroside,and swertiamarin,which are constituents of Gentianella acuta,by developing and validating a simple,sensitive,and fast ultra-hig...Objective:To simultaneously investigate the pharmacokinetics of gentiopicroside,sweroside,and swertiamarin,which are constituents of Gentianella acuta,by developing and validating a simple,sensitive,and fast ultra-high-performance liquid chromatography–tandem mass spectrometry method.Materials and Methods:Blood samples were collected from the forward limb veins of six beagle dogs following oral gavage with G.acuta,the whole plant extract(39.90 mg/kg).Plasma samples were processed using liquid–liquid extraction.The analytes and paeoniflorin(internal standard[IS])were separated using an Acquity?UPLC ethylene bridged hybrid amide column(2.1 mm×100 mm,1.7μm)with isocratic elution using a mobile phase consisting of acetonitrile and 0.1%formic acid in water(80:20,v/v)at a flow rate of 0.4 mL/min.Quantification was performed using multiple reaction monitoring of the fragmentation transitions at m/z 401.1→179.0,403.1→195.0,419.1→179.0,and 525.2→449.1 for gentiopicroside,sweroside,swertiamarin,and the IS,respectively.Results:The linearity of the analytical response was good and the calibration curves were linear over concentration ranges of 1.20–192.0,0.40–159.0,and 0.20–209.3 ng/mL for gentiopicroside,sweroside,and swertiamarin,respectively.The extraction recovery was in the range of 84.72%–91.34%,84.58%–93.43%,and 82.75%–91.37%for gentiopicroside,sweroside,and swertiamarin,respectively.Conclusions:The method was successfully used to evaluate the pharmacokinetic parameters of gentiopicroside,sweroside,and swertiamarin in beagle dogs.展开更多
基金The authors are grateful to Yunnan Major Science and Technology Project(2019ZF003,2019FY003004)the National Key Research and Development Program of China(2017YFC1704007)the general program of applied basic research of Yunnan province(2019FB116)for partial financial support.
文摘Alstonia scholaris(L.)R.Br.,an evergreen tropical plant rich in indole alkaloids with significant physiological activity,is traditionally used to treat respiratory diseases in China.This study was conducted to establish the toxicity profile of the alkaloid extract(TA)of A.scholaris leaves in non-rodents.After oral administration of a single dose(4 g/kg.bw),a num-ber of transient symptoms,such as unsteady gait,drooling,emesis,and reddening of peri-oral mucosa,were observed,but no treatment-related mortality.A sub-chronic toxicity study with a range of doses of TA(20,60 and 120 mg/kg.bw)was conducted for a 13-week treatment period,followed by 4-week recovery observation.Except for emesis and drooling in majority of animals in 120 mg/kg.bw treatment group,no clinical changes were observed in TA-treated animals.Data from electrocardiography,bone marrow,urine,fecal,hematology and clinical chemistry analyses were comparable between TA-treated and control animals.No significant differences in the relative organ weights and histopathological characteristics were evident between the TA-treated and control groups.Accordingly,the non-observed-adverse-effect-level(NOAEL)of TA was established as 120 mg/kg.bw.Our results add further knowledge to the safety database for indole alkaloid extracts from A.scholaris with potential utility as novel drug candidates.
基金supported by the China Resources Sanjiu(Ya’an)Pharmaceutical Co.,Ltd.,Sichuan,China
文摘Shenfu Injection(SFI) is a well-defined Chinese herbal formulation that is obtained from red ginseng and processed aconite root. The main active constituents in SFI are ginsenosides and aconitum alkaloids. In this work, ginsenosides(ginsenoside Rg1, ginsenoside Rb1 and ginsenoside Rc) and aconitum alkaloids(benzoylmesaconine and fuziline) were used as the index components to explore the pharmacokinetic behavior of SFI. A selective and sensitive HPLC–MS/MS method was developed for the quantification of ginsenosides and aconitum alkaloids in dog plasma and was used to characterize the pharmacokinetics of the five index components after intravenous drip of three different dosages of SFI in beagle dogs. The pharmacokinetic properties of the index components were linear over the dose range of2–8 m L/kg.
文摘This paper describes the development and validation of a liquid chromatography–mass spectrometric assay for propafenone and its application to a pharmacokinetic study of propafenone administered as a new propafenone hydrochloride sustained-release capsule(SR-test), as an instant-release tablet(IR-reference) and as the market leader sustained-release capsule(Rythmol, SR-reference) in male beagle dogs(n ? 8). In Study A comparing SR-test with IR-reference in a crossover design T_(max) and t_(1/2) of propafenone for SR-test were significantly higher than those for IR-reference while C_(max) and AUC were lower demonstrating the sustained release properties of the new formulation. In Study B comparing SRtest with SR-reference the observed C_(max) and AUC of propafenone for SR-test(124.57140.0 ng/m L and612.07699.2 ng·h/m L, respectively) were higher than for SR-reference(78.52772.92 ng/m L and423.67431.6 ng·h/m L, respectively) although the differences were not significant. Overall, the new formulation has as good if not better sustained release characteristics to the market leader formulation.
基金supported by the National Natural Science Foundation of China,No.81600738the Youth Development Project of Air Force Medical University,No.21QNPY072(both to FF)。
文摘Retinal injury after blunt ocular trauma may directly affect prognosis and lead to vision loss.To investigate the pathological changes and molecular mechanisms involved in retinal injury after blunt ocular trauma,we established a weight drop injury model of blunt ocular trauma in male Beagle dogs.Hematoxylin-eosin staining,immunofluorescence staining,western blotting,and TUNEL assays were performed to investigate retinal injury within 14 days after blunt ocular trauma.Compared with the control group,the thicknesses of the inner and outer nuclear layers,as well as the number of retinal ganglion cells,gradually decreased within 14 days after injury.The number of bipolar cells in the inner nuclear layer began to decrease 1 day after injury,while the numbers of cholinergic and amacrine cells in the inner nuclear layer did not decrease until 7 days after injury.Moreover,retinal cell necroptosis increased with time after injury;it progressed from the ganglion cell layer to the outer nuclear layer.Visual electrophysiological findings indicated that visual impairment began on the first day after injury and worsened over time.Additionally,blunt ocular trauma induced nerve regeneration and Müller glial hyperplasia;it also resulted in the recruitment of microglia to the retina and polarization of those microglia to the M1 phenotype.These findings suggest that necroptosis plays an important role in exacerbating retinal injury after blunt ocular trauma via gliosis and neuroinflammation.Such a role has important implications for the development of therapeutic strategies.
基金National Nature Science Foundation of China (30572350)New Drug Foundation of State Administration of Traditional Chinese Medicine (DIX005A)
文摘Objective To develop and validate a simple, rapid, sensitive, and reproducible HPLC method for determination of hyperoside in plasma of dogs and for the subsequent pharmacokinetic (PK) study. Methods An accurate and reproducible HPLC-UV method was developed and validated for the determination of hyperoside in plasma of dogs, using kaempferol as internal standard. The plasma samples of dogs following ig administration of hyperoside were analyzed for the detection of quercetin after enzymatic hydrolysis treatment with combined β-glucuronidase and sulphatase. The analytes were separated on a Diamonsil C 18 column (250 mm × 4.6 mm, 5 μm). The mobile phase consisted of methanol-buffer solution (0.1 mol/L NH 4 Ac + 0.3 mmol/L EDTA-Na 2 )-acetic acid (60:40:1) and was delivered at a flow rate of 1 mL/min. The UV detector was set at 370 nm and the column temperature was maintained at 35 ℃. The sample injection volume was 20 μL. Data were collected and analyzed using the ANASTAR software. PK parameters were calculated with DAS software (2.0). Results Linear relationships were validated over the range of 0.01-1 μg/mL for hyperoside (r = 0.9997). The intra- and inter-day precision values for all samples were within 10.0%, and the accuracies of intra- and inter-day assays were within the range of 92.4%-102.4%. The validated method was successfully used to determine the hyperoside concentration in plasma of dogs for up to 12 h, after a single ig administration (25 mg/kg). The mean PK parameters for male and female dogs were as follows: C max (0.18 ± 0.05) and (0.16 ± 0.05) μg/mL, AUC 0-∞ (0.79 ± 0.34) and (0.86 ± 0.27) μg/(mL·h), t 1/2(ka) (0.89 ± 0.41) and (0.88 ± 0.28) h, respectively. Statistical analysis on the PK of hyperoside in male and female groups showed that sex had no significant impact on the PK of hyperoside (P > 0.05). Conclusion The method is able and sufficient to be used in drug PK studies of hyperoside.
基金the Colleges and Universities Natural Science Foundation of Anhui Province (KJ2009B216Z)
文摘Objective To develop an LC-MS/MS method for determining the concentration of wogonin in dog plasma and investigate the pharmacokinetics and bioavailability by different administrations of wogonin in Beagle's dogs.Methods LC-MS/MS was employed in determining the concentration of wogonin with the selected ion monitoring model after liquid-liquid extraction with ethyl acetate of dog plasma samples.The lower limit of quantification was 0.105 μg/L.Target ions were at m/z 285.0→270.0 for wogonin and 373.3→305.3 for finasteride.In a randomized,self-control,and cross-over study,six male Beagle's dogs were treated with different administration methods in three test periods.Pharmacokinetic parameters were calculated with DAS software(Ver.2.0).Results The calibration curve was linear in the range of 0.105-107.36 μg/L for wogonin in dog plasma samples.The main pharmacokinetic parameters of ig administration(native drug of 15 mg/kg and solution preparation of 5 mg/kg) and iv route were as follows:Cmax(2.5 ± 1.1),(7.9 ± 3.3),and(6838.7 ± 1322.1) μg/L,tmax(0.7 ± 0.3) and(0.3 ± 0.2) h for the both former,AUC0-t(7.1 ± 2.0),(21.0 ± 3.2),and(629.7 ± 111.8) μg.h/L.The absolute bioavailability of native and solution of wogonin were(0.59 ± 0.35)% and(3.65 ± 2.00)%,respectively.Conclusion The validated method is convenient,sensitive,and specific,and the improvement of wogonin solubility could remarkably increase the absolute bioavailability.
基金supported by the Research Project of Heilongjiang University of Chinese Medicine“Supporting Plan for Excellent Innovative Talents”(2018RCD03)Heilongjiang Provincial Science Fund Project(H2018056)+2 种基金Heilongjiang Post-doctoral Research Start Fund Project(LBH-Q16214)General Projects of NSFC(81973439,81872979,and 81803686)Research Fund of Heilongjiang University of Chinese Medicine(201504)
文摘Objective:To simultaneously investigate the pharmacokinetics of gentiopicroside,sweroside,and swertiamarin,which are constituents of Gentianella acuta,by developing and validating a simple,sensitive,and fast ultra-high-performance liquid chromatography–tandem mass spectrometry method.Materials and Methods:Blood samples were collected from the forward limb veins of six beagle dogs following oral gavage with G.acuta,the whole plant extract(39.90 mg/kg).Plasma samples were processed using liquid–liquid extraction.The analytes and paeoniflorin(internal standard[IS])were separated using an Acquity?UPLC ethylene bridged hybrid amide column(2.1 mm×100 mm,1.7μm)with isocratic elution using a mobile phase consisting of acetonitrile and 0.1%formic acid in water(80:20,v/v)at a flow rate of 0.4 mL/min.Quantification was performed using multiple reaction monitoring of the fragmentation transitions at m/z 401.1→179.0,403.1→195.0,419.1→179.0,and 525.2→449.1 for gentiopicroside,sweroside,swertiamarin,and the IS,respectively.Results:The linearity of the analytical response was good and the calibration curves were linear over concentration ranges of 1.20–192.0,0.40–159.0,and 0.20–209.3 ng/mL for gentiopicroside,sweroside,and swertiamarin,respectively.The extraction recovery was in the range of 84.72%–91.34%,84.58%–93.43%,and 82.75%–91.37%for gentiopicroside,sweroside,and swertiamarin,respectively.Conclusions:The method was successfully used to evaluate the pharmacokinetic parameters of gentiopicroside,sweroside,and swertiamarin in beagle dogs.
