Clinical comprehensive evaluation of Ginkgolide Injection in treatment of cerebral infarction / 中国中药杂志

This clinical value-oriented comprehensive evaluation of drugs was carried out in accordance with Guidelines for Management of Comprehensive Clinical Evaluation of Drugs(trial version 2021), with the qualitative and quantitative evaluation methods adopted. Based on the evidence-based medicine, epidemiology, clinical medicine, pharmacoeconomics, mathematical statistics, and health technology evaluation(HTA), the clinical value of Ginkgolide Injection was evaluated from the "6+1" dimension by giving weight to the criterion level and index level and calculating with multi-criteria decision analysis(MCDA) model and CSC v2.0. After entering the market, Ginkgolide Injection has been subjected to phase Ⅳ clinical trial, spontaneous reporting system(SRS)-based data monitoring, systematic review and Meta-analysis, acute toxicity and long-term toxicity assays, active monitoring, and RCTs, and the evidence of safety was sufficient. The results of active monitoring showed that the incidence of adverse reactions was 0.09%(rare), mainly manifested as flushing, dizziness, rash, nausea, and vomiting. According to the nested case-control study, the adverse reactions of this drug had nothing to do with the product batch, implying that the drug quality was controllable. The adverse reactions mainly resulted from the pharmacodynamic reactions. Because the drug was effective in resisting platelet aggregation, the resulting adverse reactions such as flushing, dizziness, headache, and phlebitis were caused by vasodilation. Skin rash and gastrointestinal symptoms were mainly attributed to the patients' sensitivity to drugs and their own allergic constitution. According to the sufficiency of evidence and the incidence of adverse reactions in the safety research, the safety of Ginkgolide Injection was grade A. The results of Meta-analysis showed that Ginkgolide Injection combined with conventional western medicine was superior to conventional western medicine in improving the clinical effective rate, neurological function score, and activity of daily living score of patients with cerebral infarction. The validity evidence was evaluated according to the PICO principle to be high. According to the GREAD evaluation principle, the quality of such evidence as clinical effective rate, National Institute of Health stroke scale(NIHSS), and Barthel Index(BI) was evaluated, and the results demonstrated that the evidence quality of clinical effective rate and activity of daily living score was medium. The effectiveness of Ginkgolide Injection was grade A. According to the economic report of Ginkgolide Injection, it had short-term and long-term pharmacoeconomic advantages in the treatment of ischemic stroke, and the economic evidence value was good. According to the CASP economic evaluation checklist, the overall quality evaluation results of the economic report are basically clear. To be specific, the economic evidence quality was high. Based on the comprehensive economic evidence quality and economic value, the economy of this drug was grade A. The innovation of this product was evaluated from three aspects: clinical innovation, enterprise service system innovation, and industrial innovation. Ginkgolide Injection could be used 24 h after intravenous thrombolysis for improving patients' neurological function without increasing bleeding, indicating its important clinical innovation. There were many innovations in ensuring drug supply, especially at the grass roots, drug safety, effectiveness, and reasonable price, which has provided reference for establishing enterprise philosophy, managing drug resources, developing process and technology, and determining enterprise management and marketing. Therefore, its innovation was grade A. The drug had no special medication plan in use, exhibiting good suitability for doctors, nurses, and patients. The suitability was grade B. Compared with similar drugs, its price was at a medium level, meaning good affordability, sufficient production capacity, and easy accessibility. Its accessibility was therefore grade B. This drug belonged to Chinese medicinal injection. The large-sample real-world research revealed rich human use experience, so it was grade C for the traditional Chinese medicine characteristic. According to the comprehensive evaluation, the clinical value of Ginkgolide Injection in the treatment of cerebral infarction fell into class A. It is suggested that it can be transformed into the relevant policy results of basic clinical medication management according to the procedure.

Application of near infrared spectroscopy to predict contents of various lactones in chromatographic process of Ginkgo Folium / 中国中药杂志

This study established a method for rapid quantification of terpene lactone, bilobalide, ginkgolide C, ginkgolide A and ginkgolide B in the chromatographic process of Ginkgo Folium based on near infrared spectroscopy(NIRS). The effects of competitive adaptive reweighting sampling(CARS), random frog(RF), and synergy interval partial least squares(siPLS) on the performance of partial least squares regression(PLSR) model were compared to the reference values measured by HPLC. Among them, the correlation coefficients of prediction(Rp) of validation sets of terpene lactone, bilobalide, and ginkgolide C were all higher than 0.98, and the relative standard errors of prediction(RSEPs) were 5.87%, 6.90% and 6.63%, respectively. Aiming at ginkgolide A and ginkgolide B with relatively low content, the genetic algorithm joint extreme learning machine(GA-ELM) was used to establish the optimized quantitative analysis model. Compared with CARS-PLSR model, the CARS-GA-ELM models of ginkgolide A and ginkgolide B exhibited a reduction in RSEP from 15.65% to 8.52% and from 21.28% to 10.84%, respectively, which met the needs of quantitative ana-lysis. It has been proved that NIRS can be used for the rapid detection of various lactone components in the chromatographic process of Ginkgo Folium.

Effects of ginkgolide B on Caspase-3/PTEN/Akt pathway and cell proliferation and apoptosis in hypoxia/reoxygenation cardiomyocytes / 中国胸心血管外科临床杂志

@#Objective     To investigate the effect of ginkgolide B (GB) on cysteinyl aspartate specific proteinase-3 (Caspase-3)/chromosome 10 deletion phosphatase-tension protein homologue (PTEN)/protein kinase B (Akt) pathway and cell proliferation and apoptosis in hypoxia/reoxygenation cardiomyocytes. Methods     H9C2 cells were cultured in vitro. A control group was cultured in serum-free DMEM high glucose medium at 37°C and 5% CO2 for 28 hours. The remaining groups were prepared with hypoxia/reoxygenation models. A GB low-dose group and a GB high-dose group were treated with GB pretreatment with final concentration of 50 μmol/L and 200 μmol/L respectively at 1 h before hypoxia/reoxygenation. A carvedilol group was treated with carvedilol of a final concentration of 10 μmol/L at 1 h before hypoxia/reoxygenation. The proliferation and apoptosis of H9C2 cells were detected, and the levels of lactate dehydrogenase (LDH), malondialdehyde (MDA), reactive oxygen species (ROS), PTEN, Akt, phosphorylated Akt (p-Akt) and Caspase-3 in H9C2 cells were also detected. Results     Compared with the control group, the proliferation rate of H9C2 cell, and the levels of PTEN, Akt and p-Akt in other groups decreased, and the apoptosis rate, and the levels of LDH, MDA, ROS and Caspase-3 increased (P<0.05). Compared with the hypoxia/reoxygenation group, the proliferation rate of H9C2 cell, and the levels of PTEN, Akt and p-Akt in all GB dose groups and the carvedilol group increased; the apoptosis rate, and the levels of LDH, MDA, ROS and Caspase-3 decreased, and the effect of GB was in a dose dependent manner; however, the effect of GB was not as strong as carvedilol (P<0.05). Conclusion     GB can inhibit H9C2 cell apoptosis and promote H9C2 cell proliferation by activating Caspase-3/PTEN/Akt pathway.

The molecular mechanism of ginkgolide B regulating the expression of long-chain fatty acid metabolism-related proteins and antioxidant therapy for non-alcoholic fatty liver disease / 药学学报

This study investigated the effects of ginkgolide B on the long-chain fatty acid metabolism-related enzyme protein peroxisome proliferators-activated receptors α (PPARα), long-chain specific acyl-CoA dehydrogenase (LCAD), carnitine palmitoyl transterase-1 (CPT-1), and acyl coenzyme A oxidase 1 (ACOX1) expression in the liver of rats with non-alcoholic fatty liver disease (NAFLD). All the animal welfare and experimental procedures are in accordance with the regulations of the Animal Ethics Committee of Yunnan University of Traditional Chinese Medicine. After successfully building the rat model of non-alcoholic abnormal liver disease, the rats were divided into the model group, the simvastatin group, and the low-dose, middle-dose, and high-dose groups of ginkgolide B according to random number method, and were given corresponding drug treatment 4 weeks. We detected liver pathological indicators and determined blood lipids, transaminase and anti-oxidation indexes. Western blot and RT-PCR assays were used to detect the protein and mRNA levels of PPARα, LCAD, CPT-1, and ACOX1 in livers. The results showed that: ① the liver histopathology showed that the liver slices of the model group had obvious structural disorder, the nucleus was squeezed, and there were obvious fat vacuoles. The treatment groups improved significantly compared with the model group; ② compared with the normal group, the liver function and blood lipid indexes of the model group increased significantly, while the anti-oxidation indexes decreased significantly. Compared with the model group, each treatment groups were significantly improved; ③ compared with the normal group, the protein and mRNA expression levels of PPARα, ACOX1, CPT-1, and LCAD in the model group were significantly reduced, compared with the model group, those indexes in the treatment groups were significantly up-regulated. This study found that ginkgolide B could regulate the expression of long-chain fatty acid metabolism-related proteins PPARα, ACOX1, CPT-1, and LCAD, meanwhile improve the body's antioxidant capacity, thereby reduce blood lipids, further improve liver function and protect the liver.

Preparation and quality evaluation of orodispersible film containing ginkgolide B novel nanosuspension lyophilized powder / 中国中药杂志

To prepare a new dosage form that can improve the drug loading of the film--ginkgolide B nanosuspension lyophilized powder orodispersible film(GB-NS-LP-ODF) and to evaluate its quality. Firstly, ginkgolide B nanosuspension(GB-NS) was prepared by media milling method, and then ginkgolide B nanosuspension lyophilized powder(GB-NS-LP) was prepared with freeze-drying method. The mannitol was used as lyoprotectant and its dosage was also investigated. GB-NS-LP-ODF was prepared by solvent casting method and its formulation was screened by single factor test method and optimized by orthogonal test. The appearance, mechanical properties, content uniformity and in vitro dissolution of the optimized GB-NS-LP-ODF were investigated. The particle size of prepared GB-NS was about 201 nm, and the optimal dosage of mannitol was 8%. According to the optimal formula, the GB-NS-LP-ODF was prepared with GB-NS-LP 35.6%, PVA 0588 49.4%, PEG 400 10.7% and CMS-Na 4.3%, and completely disintegrated in about 30 s, and the particle size of reconstituted GB nanoparticles from ODF was about 210 nm. The film with smooth appearance and good mechanical properties was stable within 30 days and the content uniformity(A+2.2 S<15) conformed to the regulations. Scanning electron microscope(SEM) showed that GB-NS-LP-ODFs were evenly distributed and the particle size was about 200 nm. X-rays diffraction(XRD) showed that its crystallinity was significantly lower than that of GB raw drug and GB-ODF. The results of in vitro release test showed that the drug film was completely dissoluted within 10 minutes. These results indicated that nanosuspension lyophilized powder was prepared by freeze drying of nanosuspensions, and then loaded into the orodispersible film to effectively increase the drug loading of the ODF and have broad application prospects.

Clinical pharmacokinetics of Ginkgolides Dropping Pills in healthy subjects / 中草药

Objective: In order to describe the pharmacokinetic profiles of two effective constituents ginkgolide A and ginkgolide B in healthy subjects, and to provide supports for setting out the clinical application of Ginkgolides Dropping Pills. Methods: Ten healthy subjects were enrolled in a randomized and open experimental design. Following a single oral administration of Ginkgolides Dropping Pills, blood samples which were anticoagulated by heparin sodium were collected at predetermined time, and then centrifuged to separate plasma samples. The total concentration of ginkgolide A and ginkgolide B in plasma samples and the lactone concentration of ginkgolide A and ginkgolide B were determined by a verified LC-MS/MS method, the pharmacokinetic parameters were calculated by WinNonlin 6.3 with non-compartment model. Results: After a single oral administration of Ginkgolides Dropping Pills, the tmax of lactone, total concentration of ginkgolide A respectively were (3.05 ± 1.40), (3.40 ± 1.22) h, the Cmax were (84.3 ± 32.8), (92.2 ± 35.0) ng/mL, respectively, and its Cmax ratio was 91.4%. The AUC0-t were (636 ± 183), (753 ± 205) ng∙h/mL, respectively, and its AUC0-t ratio was 84.5%, half-life time (t1/2) were (13.0 ± 10.3), (12.9 ± 8.49) h, respectively. The Tmax of lactone, total concentration of ginkgolide B were (3.15 ± 1.42), (3.35 ± 1.25) h, The Cmax were (74.1 ± 31.5), (148 ± 60.1) ng/mL, respectively, and its Cmax ratio was 50.1%.The AUC0-t were (627 ± 202), (1410 ± 431) ng∙h/mL, respectively, and its AUC0-t ratio was 44.5%, t1/2 were (13.2 ± 5.83), (13.7 ± 5.83) h, respectively. Conclusion: The results demonstrated that ginkgolide A and ginkgolide B were both at a moderate absorption and elimination rate, ginkgolide A mainly existed in human plasma upon lactone, while ginkgolide B presented as hydrolyzed forms with one or two lactone groups hydrolyzed and lactone, and the two forms of ginkgolide B were at equal exposure level after single oral administration of Ginkgolides Dropping Pills.

Protective Effect and Mechanism of Ginkgolide B on Amyotrophic Lateral Sclerosis Cell Model / 中国实验方剂学杂志

Objective:To investigate the effect of ginkgolide B （GB） on the activation of c-Jun aminoterminal kinase（JNK） signaling pathway and apoptosis in amyotrophic lateral sclerosis cell model. Method:NSC34 cells were infected by slow virus containing expression superoxide dismutase1（SOD1）WT and hSOD1G93A and empty plasmid， and screened with a certain concentration of puromycin， so as to observe the transfection efficiency of slow virus and cell morphology under inverted fluorescence microscope. Western blot method was used to verify whether infected cells were over-expressing SOD1 target proteins. The hSOD1G93A-NSC34 cell lines were established and given GB. Cell cultures were divided into normal group， model group and different concentrations of ginkgolide B groups （25， 50， 75， 100 mg∙L-1）. After 48 h， methyl thiazolyl tetrazolium （MTT） was used to detect cell survival rates， and select the best drug concentration. Subsequent experimental groups were divided into normal group， model group， 75 mg∙L-1 GB group， SP600125 group， and 75 mg∙L-1 GB + SP600125 group. Flow cytometry was used to detect the apoptosis of each group of cells. Western blot was used to detect the expressions of phosphorylation（p）-JNK， c-Jun， p-c-Jun， and cysteine aspartic acid protease -3（Caspase-3） proteins. Result:Compared with normal NSC34 cells， hSOD1G93A-NSC34 cell body became round， the synapses decreased and shortened， but the cell morphology of hSODWT-NSC34 cell and empty plasmid group did not change significantly. Western blot showed that hSOD1G93A-NSC34， hSOD1WT-NSC3 intracellular SOD1 protein levels increased significantly （P<0.01）， and the amyotrophic lateral sclerosis cell model was established. Compared with the normal group， the cell activity in the model group was significantly reduced （P<0.01）. Compared with the model group， the cell activity increased at different concentrations of GB， especially when the drug concentration was 75 mg∙L-1 （P<0.01）. In subsequent experiments， compared with the normal group， the apoptosis， and expressions of p-JNK， p-c-Jun， and cleaved Caspase-3 proteins in the model group increased significantly （P<0.01）. Compared with the model group， the apoptosis and p-JNK， p-c-Jun， released Caspase-3 protein expressions of 75 mg∙L-1 GB group， SP600125 group， 75 mg∙L-1 GB + SP600125 group decreased significantly （P<0.05， P<0.01）. Conclusion:GB has a protective effect on the cell model of atrophy lateral sclerosis， which may be realized by JNK signal pathway.

Cost-minimization Analysis of Ginkgolide Injection versus Butylphthalide Injection in the Treatment of Ische- mic Stroke of Large-artery Atherosclerosis / 中国药房

OBJECTIVE：To eval uate therapeutic effic acy，safety and economical efficiency of Ginkgolide injection versus Butylphthalide injection in the treatment of ischemic stroke. METHODS ：Among the GISAA of Ginkgolide injection in the treatment of ischemic stroke of large-artery atherosclerosis ，106 patients who were given Ginkgolide injection+Asprin enteric-coated tablets but did not use butylphthalide in any dosage in previous trial group were selected as ginkgolide group ；56 patients who were given Butylphthalide injection+Ginkgolide injection+Asprin enteric-coated tablets in previous placebo group were selected as control group. The effects ，safety and economical efficiency were compared between 2 groups. Effect indexes included recurrence rate ， mortality，NIHSS score ，modified Rankin score （mRS），Barthel index and comprehensive efficacy. The safety indexes included incidence of bleeding event and adverse event during treatment. Cost-minimization analysis was used for economic evaluation. RESULTS：There was no statistical difference in recurrence rate ，mortality，NIHSS score ，the proportion of subjects with mRS 0-2，Barthel index ，comprehensive efficacy and the incidence of adverse event between 2 groups on 28th day after treatment （P＞ 0.05）. NIHSS score of ginkgolide group was better than that of control group on 7th and 14th day （P＜0.05）. Results of cost-minimization analysis showed that total cost of ginkgdide group was （13 768.19±4 981.54）yuan on 14th day of treatment ， which was significantly lower than （22 578.52±7 523.23）yuan of control group （P＜0.01）. The results of sensivity analysis indicated that the minimum lost analysis was stable. CONCLUSIONS ：For the treatment of ischemic stroke ，ginkgolide+aspirin is similar to butylphthalide+aspirin in improving clinical outcome and safety of 28 days，but is better than it in short-term efficacy of improving neurological deficit ， and better short-term economical efficiency.

Comparative study on the effects of different proportions of ginkgo ketoester and donepezil on anti-dementia based on principal component analysis and multi-attribute comprehensive index methods / 药学学报

The combination of ginkgo ketoester tablet - donepezil (GD) is a popular combination commonly used in clinic for the treatment of Alzheimer's disease. To evaluate the learning and memory improving ability of different proportions of the two drugs. We optimized the ratio of GD for treatment of dementia using a mouse model. Dementia was induced by multiple neuronal damages in mice. The experimental protocols were approved by the Animal Experimental Ethical Committee of Nanjing University of Chinese Medicine and all the procedures were strictly conducted in accordance with ethical principle of animal use and care. Morris water maze, brain hematosylin-eosin staining and the changes of the neurotransmitters and related enzymes in the plasma or brain tissues were tested to determine the effect of GD on dementia mice. The results showed that the dementia mice were significantly different from the normal group in terms of behavior, pathological sections and related indicators. Compared to the dementia mice, partial administration groups could improve learning and memory ability as well as indexes in the blood and brain tissues. Both the principal component analysis and multi-attribute comprehensive index methods were used to comprehensively evaluate the total effect of GD on anti-dementia. The results showed that the combination of two drugs at the dose of 0.5 to 1 times was in a dose-effect relationship, and the dose of 1 (the clinical equivalent) had the best treatment effect. Then based on the optimal dose, GD 1∶1 had best effect, which was consistent with the clinical use of two drugs. This provides scientific basis for more effective application of the compatibility between ketoester tablet and donepezil for modern clinic medicine.

Protective effects of ginkgolide k on focal cerebral ischemia reperfusion injury in rats on the basis of amelioration of mitochondrial calcium uniporter / 中国药学杂志

OBJECTIVE: To explore whether the protective mechanism of ginkgolide K on cerebral focal ischemia reperfusion injury in rats induced by middle cerebral artery occlusion (MCAO) was associated with the amelioration of mitochondrial calcium uniporter ( MCU) or not. METHODS: Sprague Dawley (SD) rats were divided into 5 big groups randomly: sham operation group, MCAO group, GK added into RR group, GK group and GK added into SM group. The MCAO rat model were established after cerebral artery ischemia for 2 h and reperfusion for 22 h. Zea Longa 5 score system was used to evaluate neurological deficit score; Determination of brain water content and cerebral infarction areas were determined using gravimetric method and by triphenyltetrazolium chloride(TTC) staining method, respectively. In addition, malondialdehyde (MDA) and superoxide dismutase (SOD), nitric oxide synthase (NOS), nitric oxide (NO) were detected by Elisa. Additionally, mitochondrial[Ca2+] i concentration was estimated with the fluorescence spectrophotomete. The morphological change of the injured brains were observed by HE staining. The expression of caspase-3/8/9 protein were detected by Western blot. RESULTS: Compared with GK group, GK + RR group relieved obviously the neurological deficit score and reduced the cerebral infarction areas, brain water content, mitochondrial[Ca2+]i concentration and MDA, caspase-3/8/9 protein expression while enhance SOD activity. However, the effect of SM on the GK protective activity in MCAO rat injury was the opposite in comparison to GK + RR group. CONCLUSION: The stimulative effect of RR and the inhibitory effect of SM on the GK protection in MCAO rat had proves that the protective mechanism of GK on MCAO rat injury is associate with its down-regulation of the transport capacity of MCU, leading the attenuation of mitochondrial[Ca2+]i influx.

Preparation and characterization of ginkgolide K-loaded PLGA-PEG nanoparticles and its neuroprotective activity in vitro / 中草药

Objective To prepare and characterize ginkgolide K-loaded mPEG-PLGA [poly (D,L-lactide-co-gly-colide)-block-poly (ethylene glycol)] polymer nanoparticles (GK-mPEG-PLGA-NPs) and to evaluate its neuroprotective effect on the H2O2-induced PC12 cells injury in vitro. Methods The PLGA-PEG-COOH polymer was selected as carrier and double emulsion solvent evaporation technique was employed to prepare the stealth nanoparticles. The encapsulation efficiency (EE) and drug load (DL) of GK-mPEG-PLGA-NPs were investigated by HPLC. The size distribution, zeta potential, and surface morphology of GK-mPEG-PLGA-NPs were characterized by dynamic light scattering (DLS) and transmission electron microscopy (TEM), respectively. The in vitro release of GK-mPEG-PLGA-NPs was examined using phosphate buffer solution (pH 7.4) as the releasing medium for 24 h. The H2O2-induced PC12 cells injury models was established for the investigation of the protective effect of GK-mPEG-PLGA-NPs on nerve cells in vitro. Results EE and DL of GK-mPEG-PLGA-NPs was (83.40 ± 2.85)% and (3.26 ± 0.24) mg/g, respectively. The average diameter of GK-mPEG-PLGA-NPs was (93.19 ± 2.77) nm and zeta potential was (-11.93 ± 1.71) mV. The cumulative rate of drug release was (90.5 ± 4.0)% after 60 h in phosphate buffer solution. GK-mPEG-PLGA-NPs significantly inhibited the apoptosis of PC12 cells and the release of lactic dehydrogenase induced by H2O2. However, the protective action of GK-mPEG-PLGA-NPs on the H2O2-iduced PC12 cells injury was significantly weaker than that of GK. Conclusion Our results proved that GK-mPEG-PLGA-NPs had a sustained release behavior in vitro and the neuroprotective effect of GK-mPEG-PLGA-NPs on H2O2-induced PC12 cells, which indicates that GK-mPEG-PLGA-NPs has the prospect of application and deserves further research. Key words: ginkgolide K; mPEG-PLGA; in vitro release; in vitro neuroprotection; d

Optimization of freeze-dried nanoparticles of ginkgolide B by central composite design-response surface methodology and their in vitro release studies / 中草药

Objective: In order to solve the problem of ginkgolide B (GB) for preparing the intravenous injection dosage form, such as it’s low solubility, rapid elimination in vivo and poor stability in long-term storage, a novel biodegradable polysaccharide polymer was used as a carrier to encapsulate GB into nanoparticles with sustained-release profile, which then was freeze dried for keep its stability. Methods: GB nanoparticles (GB-NP) were prepared by coacervation method with hydrophilicity polymer. The Design-Expert 8.0 software was applied for experimental design. The formulation was finalized by investigating the concentration of GB, the mass ratio of GB to polymer, and the pH of the polymer solution with size and polydispersity indexas the evaluation parameters. The optimum formulation and technique were selected by response surface method. Then the nanoparticle suspension was further freeze dried and in vitro release behavior was tested in PBS containing 30% ethanol. Results: The optimum prescription conditions were as follows: the concentration of GB was 1.5 mg/mL, the ratio of GB mass to polymer mass was 0.1 and the polymer solution pH was set at 5.0. The entrapment efficiency was (99.64 ± 0.45)% with the drug loading ratio of (9.04 ± 0.04)%. Average particle size of GB-NP was (192.8 ± 2.8) nm with a preferable PDI of 0.18 ± 0.03. The freeze-drying conditions were described as follows: using 1% mannitol as freeze- drying supporting agent, the solution of GB-NP was pre-frozen for 12 h at −80 ℃ and dried for 24 h at −40 ℃. The release behavior of free GB was rapid with the cumulative release rate reaching (64.74 ± 3.95)% during the first hour, while the cumulative release rate of GB in nanoparticles was (36.90 ± 1.41)% during the first hour. Conclusion: The GB-NP using this special biodegradable polysaccharide polymer can improve GB’s dissolution behavior in water and make GB sustained release in vitro, which is beneficial for preparing GB’s intravenous injection dosage form.

Ginkgolide A reduces airway inflammation in a neutrophil-predominant murine model of asthma via suppression of immune response of helper T cell 17 / 中国中西医结合急救杂志

Objective To determine the effects of ginkgolide A (GA) in a neutrophil-predominant murine model of asthma and explore underlying mechanisms. Methods Thirty-five female BALB/c mice were randomly divided into sham operation group (Sham group), asthma group, dexamethasone intervention control group (DEX group), low dose GA intervention group (L-GA group) and high dose GA intervention group (H-GA group), with 7 mice in each group. The asthma model was induced by intraperitoneal injection of 20 μg ovalbumin (OVA) and 75 μL Fluorine complete adjuvant (FCA) on day 0, 14 and 21, and challenged 30 minutes with 5% OVA atomization on days 22-24 consecutively; phosphate buffer (PBS) was sensitized and stimulated in Sham group. The mice in L-GA group and H-GA group were intraperitoneally injected with GA of 40 mg/kg and 80 mg/kg at 1 hour before each challenge, while the mice in DEX group were intraperitoneally injected with dexamethasone of 1 mg/kg. After 24 hours of the last OVA stimulation, the airway resistance was measured at the time of 0, 3, 6, 12, 25, 50 g/L acetylmethacholine aerosol stimulation. The total number of cells and cell classification in bronchoalveolar lavage fluid (BALF) were counted. The transforming growth factor-β1 (TGF-β1) and interleukin-17 (IL-17) in BALF were detected by enzyme linked immunosorbent assay (ELISA). The proportion of helper T cell 17 (Th17) to CD4+ T cell in lung tissue was detected by flow cytometry, and the pathological characteristics of lung tissue were evaluated. Results Compared with the Sham group, the airway hyper responsiveness (AHR), the total cells, the neutrophil counts, the levels of TGF-β1, IL-17 in BALF, and the proportion of Th17 cells in the lung tissue in the asthma group were significantly increased, obvious inflammatory cell infiltration and collagen fiber deposition around airway were observed, and airway inflammation score and mucus score were significantly increased. Compared with the asthma group, low and high doses of GA significantly reduced AHR, and there was a significant difference in airway resistance at the time of 50 g/L acetylmethacholine stimulation (cmH2O·s-1·mL-1: 5.29±0.40, 3.99±0.57 vs. 7.34±0.77, both P < 0.05); the total cells, neutrophil counts, and levels of TGF-β1, IL-17 in BALF, and the proportion of Th17 cells in lung tissue were significantly decreased [total cells count (×104/L): 21.00±1.00, 17.00±1.02 vs. 27.50±2.50; neutrophil count (×104/L): 12.600±0.600, 10.610±0.210 vs. 16.875±1.125; TGF-β1 (ng/L): 371.40±107.80, 289.60±70.76 vs. 551.90±68.34; IL-17 (ng/L): 60.75±11.79, 44.77±7.09 vs. 122.50±38.87; the proportion of Th17 cells: (5.53±0.40)%, (3.76±1.10)% vs. (8.30±1.19)%, all P < 0.05]; inflammatory cell infiltration around the airway and mucus secretion was significantly reduced, airway inflammation score and mucus score were significantly decreased (2.16±0.28, 1.16±0.28 vs. 3.77±0.25; 1.33±0.58, 1.17±0.29 vs. 3.67±0.58, all P < 0.05). The AHR, total cells, neutrophil counts, and IL-17 level in BALF, the proportion of Th17 cells in lung tissue and airway inflammation score decreased more obviously with the increase of GA dosage (all P < 0.05). For index mentioned above, no significant differences were observed between DEX group and asthma group. Conclusion GA treatment was effective in a murine model of neutrophil-predominant asthma via inhibiting response in the immune cells Th17.

Neuroprotective effects of Ginkgo biloba extract and Ginkgolide B against oxygen-glucose deprivation/reoxygenation and glucose injury in a new in vitro multicellular network model / 医学前沿

Acute ischemic stroke (AIS), as the third leading cause of death worldwide, is characterized by its high incidence, mortality rate, high incurred disability rate, and frequent reoccurrence. The neuroprotective effects of Ginkgo biloba extract (GBE) against several cerebral diseases have been reported in previous studies, but the underlying mechanisms of action are still unclear. Using a novel in vitro rat cortical capillary endothelial cell-astrocyte-neuron network model, we investigated the neuroprotective effects of GBE and one of its important constituents, Ginkgolide B (GB), against oxygen-glucose deprivation/reoxygenation and glucose (OGD/R) injury. In this model, rat cortical capillary endothelial cells, astrocytes, and neurons were cocultured so that they could be synchronously observed in the same system. Pretreatment with GBE or GB increased the neuron cell viability, ameliorated cell injury, and inhibited the cell apoptotic rate through Bax and Bcl-2 expression regulation after OGD/R injury. Furthermore, GBE or GB pretreatment enhanced the transendothelial electrical resistance of capillary endothelial monolayers, reduced the endothelial permeability coefficients for sodium fluorescein (Na-F), and increased the expression levels of tight junction proteins, namely, ZO-1 and occludin, in endothelial cells. Results demonstrated the preventive effects of GBE on neuronal cell death and enhancement of the function of brain capillary endothelial monolayers after OGD/R injury in vitro; thus, GBE could be used as an effective neuroprotective agent for AIS/reperfusion, with GB as one of its significant constituents.

Applying risk management to analytical methods for the desorbing process of ginkgo diterpene lactone meglumine injection / 中国天然药物

Analysis errors can occur in the desorbing process of ginkgo diterpene lactone meglumine injection (GDMI) by a conventional analysis method, due to several factors, such as easily crystallized samples, solvent volatility, time-consuming sample pre-processing, fixed method, and offline analysis. Based on risk management, near-infrared (NIR) and mid-infrared (MIR) spectroscopy techniques were introduced to solve the above problems with the advantage of timely analysis and non-destructive nature towards samples. The objective of the present study was to identify the feasibility of using NIR or MIR spectroscopy techniques to increase the analysis accuracy of samples from the desorbing process of GDMI. Quantitative models of NIR and MIR were established based on partial least square method and the performances were calculated. Compared to NIR model, MIR model showed greater accuracy and applicability for the analysis of the GDMI desorbing solutions. The relative errors of the concentrations of Ginkgolide A (GA) and Ginkgolide B (GB) were 2.40% and 2.89%, respectively, which were less than 5.00%. The research demonstrated the potential of the MIR spectroscopy technique for the rapid and non-destructive quantitative analysis of the concentrations of GA and GB.

Applying risk management to analytical methods for the desorbing process of ginkgo diterpene lactone meglumine injection / 中国天然药物

Analysis errors can occur in the desorbing process of ginkgo diterpene lactone meglumine injection (GDMI) by a conventional analysis method, due to several factors, such as easily crystallized samples, solvent volatility, time-consuming sample pre-processing, fixed method, and offline analysis. Based on risk management, near-infrared (NIR) and mid-infrared (MIR) spectroscopy techniques were introduced to solve the above problems with the advantage of timely analysis and non-destructive nature towards samples. The objective of the present study was to identify the feasibility of using NIR or MIR spectroscopy techniques to increase the analysis accuracy of samples from the desorbing process of GDMI. Quantitative models of NIR and MIR were established based on partial least square method and the performances were calculated. Compared to NIR model, MIR model showed greater accuracy and applicability for the analysis of the GDMI desorbing solutions. The relative errors of the concentrations of Ginkgolide A (GA) and Ginkgolide B (GB) were 2.40% and 2.89%, respectively, which were less than 5.00%. The research demonstrated the potential of the MIR spectroscopy technique for the rapid and non-destructive quantitative analysis of the concentrations of GA and GB.

Effects of ginkgolide B on the ventricular wall motion of patients with acute myocardial infarction after revascularization / 中成药

AIM To investigate the role of ginkgolide B on the ventricular wall motion and systolic function in acute myocardial infarction (AMI) patients after their revascularization.METHODS A total of 80 cases of AMI patients who underwent revascularization were divided into control group and ginkgolide B treatment group,with 40 patients per group.The two groups of patients both treated basically with routine western medicine were dosed with either placebo or ginkgolide B for 3 months.The cardiac function,normal myocardial percentages,ventricular wall motion,longitudinal peak systolic strain (LPSS) and rate (LPSSR) were evaluated by dobutamine stress echocardiography and these indexes were compared.RESULTS Compared to the control group,left ventricular ejection fraction (LVEF) (66.06 ± 8.39 vs.60.45 ± 13.35,P ＜0.05) and normal myocardial percentages (86.88 ±8.76 vs.79.84 ± 12.25,P ＜0.01) were significantly improved at the 2nd week in ginkgolide B treatment group,but no significant difference on ventricular wall motion was observed'between the two groups (P ＞ 0.05).For the patients with anterior wall AMI,the minus LPSS and LPSSR at basal,middle and apex segment were significantly improved at the 2nd week in ginkgolide B treatment group compared to the control group (P ＜ 0.05,P ＜ 0.01).Moreover,LPSS at apex segment was significantly improved at the 3rd month in ginkgolide B treatment group compared to the control group (P ＜ 0.05).For the patients with inferior wall AMI,minus LPSS at basal and apex segment were significantly improved at the 2nd week and the 3rd month in ginkgolide B treatment group compared to the control group (P ＜ 0.05),whereas minus LPSSR at basal and middle segment at the 2 nd week and basal segment at the 3rd month were significantly improved in ginkgolide B treatment group compared to the control group (P ＜0.05).CONCLUSION Revascularized AMI patients treated with ginkgolide B can expect an improved ventricular wall motion.

Toxicokinetics of Ginkgolide B Injection in Beagle dogs / 医药导报

Objective To investigate the toxicokinetic properties of ginkgolide B(GB) injection after single or repeat-ed administration by intravenous drip in Beagle dogs and to provide evidence for its rational use. Methods Beagle dogs were randomly divided into three groups,and received GB injection at big,medium and small doses of 80,20 and 5 mg·kg-1, re-spectively,by iv drip for 30 min per day and for 6 consecutive days per week for up to 91 days.The blood samples of Beagle dogs were drawn at different time points on the first and last day of administration,and concentrations in plasma were detected by GC-MS method.Toxicokinetic parameters were calculated by DAS pharmacokinetic software and statistically analyzed by SPSS 11.5 software. Results The elimination half-life (t1/2β) of GB at single dose of 5,20,80 mg·kg-1were(110.2±32.6),(115.4± 12.8),(98.6±26.8) min, respectively.The AUC0-twere (61.1±7.4), (348.6±90.5), (2 046.2±356.4) mg·L-1·min,re-spectively.The t1/2βof GB at mutiple doses of 5,20,80 mg·kg-1on the 91rd day were (117.9±28.0),(118.2±17.0),(120.5± 49.4) min,respectively.The AUC0-twere (67.9±14.9), (218.3±31.8), (1 986.4±426.6) mg·L-1·min, respectively.There was no significant difference in main toxicokinetic parameters including t1/2βamong the single or repeated dosage groups, but AUC0-tand Cmaxincreased proportionally with doses. Conclusion The curves of single and repeated intravenous drip of GB in-jection in beagle dogs were in line with the two atrioventricular model,with linear dynamic characteristics and there was no accu-mulation of repeated drug delivery in the body.

Effect of compatibility of ginkgolide A, ginkgolide B and ginkgolide K / 中国中药杂志

To investigate the best active compatibility of ginkgolide A, B and K (GA，GB，GK). The effects of GA, GB, GK alone, combinations of each two of them, and combinations of these three components on platelet-activating factor (PAF)-induced platelet aggregation activity and rat cerebral ischemia reperfusion model (tMCAO) were compared in this study. Different compatibilities of GA, GB and GK could significantly reduce the maximum aggregation rate of PAF-induced platelet aggregation, and the effect was most obvious in combination of the three. Different compatibilities of GA, GB and GK could alleviate the neural function, cerebral infarction volume and cerebral edema in the tMCAO model of rats to different degrees, and the effect of combinations of the three was stronger than those of combinations of two and single use. The combination of all of GA, GB and GK had the strongest effect on nerve injury caused by anti-platelet aggregation in tMCAO rats.

Study on pharmacokinetics and pharmacodynamics of ginkgolides in rat model with cerebral ischemia injury / 中草药

Objective To study the metabolic process of ginkgolides in rats with cerebral ischemic injury based on pharmacokinetic- pharmacodynamics (PK-PD) binding model. Methods The middle cerebral artery occlusion (MCAO) model was established by the suture method. After reperfusion, rats were randomly assigned to nasal administration, ig administration, and iv administration group.The orbital blood was taken at different time points of 0.25, 0.33, 0.5, 0.75, 1.0, 1.25, 1.5, 2.0, 4.0, 6.0, 9.0, and 12.0 h after the administration of the ginkgolides solution. The drug-time curve of ginkgolide B in plasma were drawn according to the concentration measured by LC-MS. The time-effect curve of superoxide dismutase (SOD) and malondialdehyde (MDA) were drawn based on the value measured by the kit. The pharmacokinetic parameters were calculated by DAS 2.0 software to fit the PK-PD binding model. Results The t1/2 of ginkgolide B of the rats in the administration group was smaller than that in the MCAO model group. The area under the curve (AUC) of nasal administration was significantly higher than intragastric administration and intravenous administration. Conclusion Ginkgolide B has a good protective and mitigating effect on ischemic stroke. The pharmacokinetics of nasal administration is better than iv and ig administration, which can provide reference for the development of nasal administration of ginkgolide B.