Effect and Safety of Guanxinning Tablet(冠心宁片) for Stable Angina Pectoris Patients with Xin(Heart)-Blood Stagnation Syndrome:A Randomized,Multicenter,Placebo-Controlled Trial

Objective: To investigate the effect and safety of Guanxinning Tablet(冠心宁片, GXN) for the treatment of stable angina pectoris patients with Xin(Heart)-blood stagnation syndrome(XBSS). Methods: One hundred and sixty stable angina pectoris patients with XBSS were randomly assigned to receive GXN(80 cases) or placebo(80 cases, Guanxinning simulation tablets, mainly composed of lactose), 4 tablets(0.38 g/tablet), thrice daily for 12 weeks. After treatment, an exercise stress test(treadmill protocol), Chinese medicine(CM) syndrome score, electrocardiogram(ECG), and nitroglycerin withdrawal rate were evaluated and compared in the patients between the two groups. Meanwhile, adverse events(AEs) were evaluated during the whole clinical trial. Results: Compared with the control group, the time extension of exercise duration in the GXN group increased 29.28±17.67 s after treatment(P>0.05);moreover, the change of exercise duration in the GXN group increased 63.10±96.96 s in subgroup analysis(P<0.05). The effective rates of angina pectoris, CM syndrome and ECG as well as nitroglycerin withdrawal rate were 81.33%, 90.67%, 45.76%, and 70.73%, respectively in the GXN group, which were all significantly higher than those in the control group(40.58%, 75.36%, 26.92%, 28.21%, respectively, P<0.05). Conclusion: GXN was a safe and effective treatment for stable angina pectoris patients with XBSS at a dose of 4 tablets, thrice daily.

Guanxinning tablet inhibits the interaction between leukocyte integrin Mac-1 and platelet GPIbαfor antithrombosis without increased bleeding risk

Recent studies have showed that thrombosis is closely related to leucocytes involved in immunity.Interfering with the binding of leukocyte integrin Mac-1 and platelet GPIbαcan inhibit thrombosis without affecting physiological coagulation.Mac-1-GPIbαis proposed as a potential safety target for antithrombotic agents.Guanxinning tablet(GXNT)is an oral Chinese patent medicine used for the treatment of angina pectoris,which contains phenolic acid active ingredients,such as salvianolic acids,ferulic acid,chlorogenic acid,caffeic acid,rosmarinic acid,tanshinol,and protocatechualdehyde.Our previous studies demonstrated that GXN exhibited significant antithrombotic effects,and clinical studies suggested that it did not increase bleeding risk.In addition,GXN exerted a significantly regulatory effect on immune inflammation.In the current study,we intended to evaluate the effects of GXN on bleeding events and explore the safety antithrombotic mechanism of GXN based on leukocyte-platelet interaction.First,we established a gastric ulcer model induced by acetic acid in rats and found that GXN not only did not increase the degree of gastrointestinal bleeding when gastric ulcer occurred,but also had a certain promoting effect on the healing of gastric ulcer.Second,in vitroexperiments showed that after pretreatment with GXN and activation by phorbol 12-myristate-13-acetate(PMA),the adhesion and aggregation of leukocytes with human platelets were reduced.It was also found that GXN reduced the expression and activation of Mac-1 in leucocytes,and inhibited platelet activation due to leukocyte engagement via Mac-1.Overall,the results suggest that GXN may be a safe antithrombotic agent,and its low bleeding risk mechanism is probably related to inhibited leukocyte-platelet aggregation and its interaction target Mac-1-GPIbα.

Network Pharmacology-Based Exploration of the Mechanism of Guanxinning Tablet for the Treatment of Stable Coronary Artery Disease

Objective:Network pharmacology was utilized to explore the mechanism of Guanxinning(GXN)tablet for the treatment of stable coronary artery disease(SCAD).Materials and Methods:First,active ingredients and therapeutic targets were predicted by databases and gene chip.Then,we constructed the compound-target(C-T)network and target-disease(T-D)network to screen hub compounds and therapeutic targets based on contribution index(CI),degree,closeness,betweenness,and coreness in the networks.Enrichment analysis was performed on hub therapeutic targets,and finally,the verification of hub ingredients and hub therapeutic targets was performed through molecular docking.Results:With"oral bioavailability≥30%,druglikeness≥0.18,and half-life≥4 h"as screening conditions,58 active ingredients were obtained.Seven hundred and seventeen compound targets and 636 SCAD targets were retrieved using databases and gene chip,and the intersection of both(139 targets)was defined as therapeutic targets.According to CI,degree,betweenness,closeness,and coreness,2 hub compounds and 13 hub therapeutic targets were chosen from the C-T network and T-D network,respectively.The Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis indicated that GXN treated SCAD from several aspects including inflammatory reaction,oxidative stress,nutritional metabolism,blood pressure regulation,ventricular remodeling,vascular smooth muscle proliferation,angiogenesis,and platelet aggregation.Tissue enrichment analysis revealed that the therapeutic targets were enriched in multiple organs and tissues.The excellent binding force between the hub compounds and hub therapeutic targets was verified by molecular docking.Conclusions:The treatment of SCAD by GXN has the characteristics of multiple ingredients,multiple targets,and multiple approaches.Consequently,it may theoretically treat SCAD from multiple angles and levels.

Population pharmacokinetics of paeoniflorin in guanxin Ⅱ prescription

To evaluate the effect of components in Guanxin Ⅱ prescription on the pharmacokinetic profiles of paeoniflorin. Plasma concentration of Paeoniflorin in rats after intravenous injection of Paronia Pall Extract （PPE） and oral administration of PPE and three types of decoctions in Guanxin Ⅱ prescription, respectively, were determined by HPLC analyses. NONMEM （nonlinear mixed-effect modeling） method was used to analyze full set of pharmacokinetic data directly. A two-compartment model with first-order degradation in absorption compartment was employed for the data analysis. The mean of population parameters, CL1, V1, CL2, V2, Ka0, and Kal, were measured to be 0.509 L/h, 0.104 L, 0.113 L/h, 0.123 L, 0.135/h, and 0.0135/h, respectively. Inter-individual variabilities were estimated and dose formulation （DF） was identified as a significant covariate of Ka 1, Ka0, and V1. It is concluded that the pharmacokinetic behaviors of paeoniflorin in rats can alter with different dose formulations.

Simultaneous Determination of Four Ingredients in Guanxin Danshen Tablets by RP-HPLC

[Objectives]This study aimed to establish a method for simultaneous determination of four ingredients in Guanxin Danshen tablets by RP-HPLC.[Methods]HPLC chromatography was adopted with column of Thermo Hypersil-Keystone C 18 column(4.6 mm×250 mm,5μm),mobile phase of acetonitrile(A)-0.03 mol/L ammonium acetate(pH adjusted to 2.4 with formic acid)(B),gradient elution(0-5 min,5%A;5-10 min,5%A→19%A;10-40 min,19%A;40-68 min,19%A→36%A;68-90 min,36%A→95%A),flow rate of 1.0 mL/min and column temperature of 25℃.[Results]The content of salvianic acid A sodium,protocatechuic aldehyde,salvianolic acid B and tanshinone II A showed good linear relationship with chromatographic peak area in the range of 3.310-18.66,0.03950-0.2370,0.7500-4.500,0.05920-0.3550μg,respectively.The recovery rate(n=6)was 101.75%,96.86%,104.15%and 99.03%,respectively,and the RSD was 1.52%,2.81%,1.80%,and 1.37%respectively.The established method has good precision,reproducibility and stability.[Conclusions]This method can be used for the quality control of multiple ingredients of Guanxin Danshen tablets.

Effect of Guanxin Danshen Dripping Pills on Coronary Heart Disease Comorbid with Depression or Anxiety: The ADECODE-Real World Study

Objective: To evaluate the efficacy of Guanxin Danshen Dripping Pill(GXDSDP) in treating anxiety and depression in patients with coronary heart disease(CHD). Methods: A total of 1,428 patients diagnosed with CHD screened for anxiety, depression, and quality of life(QOL) at baseline received 0.4 g of GXDSDP treatment 3 times per day and returned for monthly reassessment. Patients were recruited after stable treatment for CHD and received assessment of General Anxiety Disorder-7(GAD-7), Patient Health Questionnaire-9(PHQ-9), and Seattle Angina Questionnaire(SAQ) for evaluating anxiety, depression, and QOL.Patients were followed up 3 times, once every 4 weeks, during outpatient visits for 12 weeks. Results: At the third follow-up(F3), the anxiety symptom of 63.79%(673/1,055) of the patients improved to sub-clinical level, and the GAD-7 score improved significantly(8.11 vs. 3.87, P<0.01);57.52%(585/1,017) patients' depressive symptoms improved to sub-clinical level, with a significant improvement in PHQ-9 score(8.69 vs. 4.41, P<0.01) at F3. All aspects of QOL significantly improved at the end of treatment compared to those at baseline(all P<0.01) as assessed by SAQ: physical limitation(31.17 vs. 34.14), anginal stability(2.74 vs. 4.14), anginal frequency(8.16vs. 9.10), treatment satisfaction(13.43 vs. 16.29), and disease perception(8.69 vs. 11.02). Conclusions: A fixed dosage of GXDSDP may be a potential treatment option for CHD patients comorbid with anxiety or depression.(Registration No. ChiCTR2100051523)

Absorbed Bioactive Compounds Replicate GuanxinⅡ-Induced Endothelium-Associated in/ex vivo Vasodilation

Objective:To develop an interference-free and rapid method to elucidate GuanxinⅡ(GXⅡ)'s representative vasodilator absorbed bioactive compounds(ABCs)among enormous phytochemicals.Methods:The contents of ferulic acid,tanshinol,and hydroxysafflor yellow A(FTA)in GXⅡ/rat serum after the oral administration of GXⅡ(30 g/kg)were detected using ultra-performance liquid chromatography-mass spectrometry.Totally 18 rats were randomly assigned to the control group(0.9%normal saline),GXⅡ(30 g/kg)and FTA(5,28 and 77 mg/kg)by random number table method.Diastolic coronary flow velocity-time integral(VTI),i.e.,coronary flow or coronary flow-mediated dilation(CFMD),and endothelium-intact vascular tension of isolated aortic rings were measured.After 12 h of exposure to blank medium or 0.5 mmol/L H_(2)O_(2),endothelial cells(ECs)were treated with post-dose GXⅡof supernatant from deproteinized serum(PGSDS,300μL PGSDS per 1 m L of culture medium)or FTA(237,1539,and 1510 mg/m L)for 10 min as control,H_(2)O_(2),PGSDS and FTA groups.Nitric oxide(NO),vascular endothelial growth factor(VEGF),endothelin-1(ET-1),superoxide dismutase(SOD),malondialdehyde(MDA)and phosphorylated phosphoinositide 3 kinase(p-PI3K),phosphorylated protein kinase B(p-AKT),phosphorylated endothelial nitric oxide synthase(p-e NOS)were analyzed.PGSDS was developed as a GXⅡproxy of ex vivo herbal crude extracts.Results:PGSDS effectively eliminates false responses caused by crude GXⅡpreparations.When doses equaled the contents in GXⅡ/its post-dose serum,FTA accounted for 98.17%of GXⅡ-added CFMD and 92.99%of PGSDS-reduced vascular tension.In ECs,FTA/PGSDS was found to have significant antioxidant(lower MDA and higher SOD,P<0.01)and endothelial function-protective(lower VEGF,ET-1,P<0.01)effects.The increases in aortic relaxation,endothelial NO levels and phosphorylated PI3K/Akt/e NOS protein induced by FTA/PGSDS were markedly abolished by NG-nitro-L-arginine methyl ester(L-NA,e NOS inhibitor)and wortmannin(PI3K/AKT inhibitor),respectively,indicating an endothelium-dependent vasodilation via the PI3K/AKT-e NOS pathway(P<0.01).Conclusion:This study provides a strategy for rapidly and precisely elucidating GXⅡ's representative in/ex vivo cardioprotective absorbed bioactive compounds(ABCs)-FTA,suggesting its potential in advancing precision ethnomedicine.

Clinical efficacy and gene chip expression analysis of Shenzhu Guanxin recipe granules(参术冠心方颗粒) in patients with intermediate coronary lesions

OBJECTIVE: To evaluate the clinical efficacy and safety of Shenzhu Guanxin recipe granules(参术冠心颗粒, SGR) in treating patients with intermediate coronary lesions(ICL), and to investigate the potential mechanism though a transcriptome sequencing approach. METHODS: ICL patients with Qi deficiency and phlegm stasis were adopted and randomly assigned to a case group or a control by random number generator in a 1∶1 randomization ratio to evaluate the clinical efficacy. RESULTS: There was no significant difference between the two groups in coronary computed tomography angiography related indexes in the two groups before and after intervention. Through the gene chip expression analysis, it is finally concluded that there are 355 differential mRNAs(190 up-regulated genes and 165 down regulated genes) when compared the SGR group and placebo group. Through protein-protein interaction network analysis of differentially expressed genes, 10 hub genes were finally obtained: CACNA2D2, CACNA2D3, DNAJC6, FGF12, SGSM2, CACNA1G, LRP6, KIF25, OXTR, UPB1. CONCLUSIONS: SGR combined with Western Medicine can be safely used to treat ICL patients with Qi deficiency and phlegm stasis. The possible mechanism of action and relevant gene loci and pathway were proposed.

GuanxinⅤRelieves Ventricular Remodeling by Inhibiting Inflammation:Implication from Virtual Screening,Systematic Pharmacology,Molecular Docking,and Experimental Validation

Objective:To reveal the anti-inflammatory mechanism of GuanxinⅤ,which is prescribed for ventricular remodeling in clinical practice.Methods:GuanxinⅤ-,ventricular remodeling-,and inflammationrelated targets were obtained through an integrated strategy of virtual screening and systematic pharmacology,and then the shared targets were visualised with a Venn diagram.GuanxinⅤnetwork and the protein-protein interaction network were drawn,and enrichment analysis was conducted.Finally,the main results obtained from the integrated strategy were validated by molecular docking and in vivo experiments.Results:A total of 251,11,425,and 15,246 GuanxinⅤ-,ventricular remodeling-,and inflammation-related targets were acquired,respectively.Then,211 shared targets were considered to contribute to the mechanism of ventricular remodeling treated by GuanxinⅤ.Guanxin network and the protein-protein interaction network were drawn,and enrichment analysis showed some cardiovascular-related biological processes and signaling pathways.Molecular docking revealed that the GuanxinⅤ-derived compounds could align with key targets.Final in vivo experiments proved that GuanxinⅤreverses ventricular remodeling by inhibiting inflammation.Conclusion:GuanxinⅤrelieves ventricular remodeling by regulating inflammation,which provides new ideas for the anti-ventricular remodeling mechanism of GuanxinⅤ.

Effectiveness of Shenshu Guanxin recipe granules(参术冠心方颗粒)for improving exercise tolerance in patients with stable angina pectoris:a randomized,double-blind,placebo-controlled trial

OBJECTIVE:To assess the effectivess of Shenshu Guanxin recipe granules(参术冠心方颗粒,SGR)in improving exercise tolerance and the quality of life in patients with Stable Angina Pectoris(SAP).METHODS:A total of 189 patients were consecutively enrolled between December 2012 and December 2014.The included patients were randomly assigned to SGR and placebo groups.The primary endpoints included mainly the results of treadmill exercise test and Seattle Angina Questionnaire(SAQ)during 12 weeks of treatment.RESULTS:After 12 weeks of treatment,SGR extended the time of exercise-induced ST-segment depression of 0.1 MV,lowered the maximum ST-segment depression,and shortened the duration of ST-segment depression in patients with SAP in southern China.Besides,the study also proved that SGR could improve the quality of life and functional status of patients with SAP.CONCLUSIONS:SGR showed a positive effect on exercise tolerance compared with the placebo besides optimal medical therapy.Also,the study proved that SGR could improve the SAQ score of the patients.

Effect of Shenzhu Guanxin Recipe (参术冠心方)on Patients with Angina Pectoris after Percutaneous Coronary Intervention:A Prospective, Randomized Controlled Trial

Objective： To evaluate the efficacy and safety of a combination therapy using Chinese medicine （CM） Shenzhu Guanxin Recipe （参术冠心方, SGR） and standard Western medicine treatment （SWMT） in patients with angina pectoris after percutaneous coronary intervention （PCI）. Methods： Double-blind randomized controlled trial was used in this experimental procedure. One hundred and eighty-seven patients with coronary heart disease receiving SWMT after PCI were randomly assigned to the treatment （SGR） and control （placebo） groups. Outcome measures including angina pectoris score （APS）, CM symptom score, and Seattle Angina Questionnaire （SAQ） score were evaluated in 1, 2, 3 and 12 months, and the death rate, restenosis and other emergency treatments were observed. The mixed-effects models were employed for the data analysis. Results： In the treatment group, a larger within-treatment effect size （d=1.74） was found, with a 76.7% reduction in APS from pretreatment to 12-month follow-up assessment compared with the control group （d=0.83, 53.8% symptom reduction）; between- treatment （BT） effect size was d=0.66. CM symptom scores included an 18.3% reduction in the treatment group （d=0.46）, and a 16.1% decrease in the control group （d=0.31）; d=0.62 for BT effect size. In regard to scores of SAQ, the BT effect size of cognition level of disease was larger in the treatment group （d=0.63）, followed by the level of body limitation of activity （d=0.62）, condition of angina pectoris attacks （d=0.55）, satisfaction level of treatments （d=0.31）, and steady state of angina pectoris （d=0.30）. Two cardiovascular related deaths and one incidental death were recorded in the control and treatment groups, respectively. No significant difference in any cardiovascular event （including death toll, frequency of cardiovascular hospitalization or emergency room visits） was found between the two groups. Conclusion： The combination therapy of SGR and SWMT is effective and safe in patients with angina pectoris after PCI when compared with SWMT alone.

Network Pharmacology-Based Exploration of Synergistic Mechanism of Guanxin Ⅱ Formula (冠心Ⅱ号方) for Coronary Heart Disease

Objective:To study the pharmacological mechanism of Guanxin Ⅱ formula(冠心Ⅱ号方,GXⅡ)for treatment of coronary heart disease(CHD).Methods:A network pharmacology-based method was utilized.First candidate compounds,targets of GXⅡ were collected using Pharm Mapper,BATMAN-TCM,Drug Bank and Swiss Target Prediction,and targets on CHD were mined from Gene Cards,Dis Genet,Drug Bank and GEO.Afterwards,the big hub compounds and targets were chosen in the candidate compounds-direct therapeutic targets on the CHD(C-T)network and the direct therapeutic targets on the CHD(T-D)network.Furthermore,the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis were performed to identify the enriched terms.Finally,a molecular docking simulation strategy was adopted to verify the binding capacity between the big hub compounds and big hub targets on CHD.Results:First,114 candidate compounds were selected with the following criteria:OB 30%,DL 0.18,and HL 4 h.Then,1,035 targets of GXⅡ were gathered,while 928 targets on CHD were collected.Afterwards,196 common targets of compound targets and therapeutic targets on CHD were defined as direct therapeutic targets acting on CHD.In addition,the contribution index(CI)in the C-T network was calculated,and 4 centrality properties,including degree,betweenness,closeness and coreness,in the T-D network,4 big hub compounds,and 6 big hub targets were eventually chosen.Furthermore,the GO and KEGG analysis indicated that GXⅡ acted on CHD by regulating the reactive oxygen species metabolism,steroid metabolism,lipid metabolism,inflammatory response,proliferation,differentiation and apoptosis.The docking results manifested excellent binding capacity between the 4 big hub compounds and 6 big hub targets on CHD.Conclusion:This network pharmacology-based exploration revealed that GXⅡ might prevent and inhibit the primary pathological processes of CHD.

Guanxin Ⅱ (冠心Ⅱ号) for the Management of Coronary Heart Disease

This article presents an integrated overview of GuanxinⅡ(冠心Ⅱ号)regarding its quality control,pharmacokinetics,pharmacology,clinical studies,adverse events,dosage and administration,and its pharmacoeconomic assessment.It has been demonstrated that GuanxinⅡhas beneficial effects on coronary heart disease(CHD).The underlying mechanism was proved to be its anti-ischemic,anti-apoptotic,antioxidative, antiplatelet and anti-inflammatory effects,and so on.Tanshinol,hydroxysafflor yellow A and ferulic acid might be responsible for the cardioprotective effect of GuanxinⅡ.In terms of acquisition cost,GuanxinⅡis cheaper than other drugs currently available for CHD.GuanxinⅡis safe,cheap,and effective in the management of CHD.However,the mechanism of its cardioprotective effects has not been completely understood because of limitations in the research methodologies of Chinese medicine.Further work should be carried out with single components such as tanshinol,hydroxysafflor yellow A and ferulic acid,using modern biochemical and molecular methods.

Shenzhu Guanxin Recipe Granules (参术冠心方颗粒) for Improving Exercise Tolerance in Patients with Stable Angina (SERIES Trial):A Protocol of Multicenter,Randomized,Double-Blind,Placebo Parallel Controlled Clinical Trial

Background: Many patients with chronic angina experience anginal episodes despite successful recanalization, antianginal and antiischemic medications. Empirical observations suggested that Shenzhu Guanxin Recipe Granules(参术冠心方颗粒, SGR), a Chinese herbal compound, exerted potential impacts on increased treadmill exercise performance and angina relieve. However, there has been no systematic study to clarify the impact of SGR on exercise tolerance in patients with stable angina. The SERIES(ShEnzhu guanxin Recipe for Improving Exercise tolerance in patients with Stable angina) trial is designed to determine the effects of SGR on exercise duration, electrocardiographic(ECG) evidence of myocardial ischemia, and incidence of major adverse cardiac events(MACE) in stable anginal patients. Methods: A total of 184 eligible patients with stable angina will be randomly assigned to receive placebo or SGR(10 g/day for 12 weeks) in a 1:1 ratio. The primary outcome will be the change from baseline in total exercise tolerance duration, time to onset of angina and ECG ischemia during exercise treadmill testing performed over a 12-week study period. The secondary outcome will include ECG measures, the occurrence and composite of MACE and the Seattle Angina Questionnaire score. Moreover, the coronary microcirculation will be evaluated to explore the possible effects in response to treatment of SGR. After the procedure, all participants will be followed up by interview at 3 and 6 months, enquiring about any cardiac events, hospitalizations, cardiac functional level and medication usage. Additionally, the occurrence of adverse events will be evaluated at each follow-up. Discussion: This study may provide novel evidence on the efficacy of SGR in improving exercise tolerance and potentially reducing clinical adverse events.