Study On The Anti-myocardial Ischemia Effect And Pharmacokinetics Of PF₁₁ Pills

Pseudoginsenoside F11(PF11),the characteristic component distinguishing the American ginseng from Chinese ginseng,is chosen as drug substance to make an innovative anti-myocardial ischemia drug.The drug product,PF11 Pill,is registated as Chemicals 2.1.In this paper,the anti-myocardial ischemia effect,pharmacodynamics mechanism and pharmacokinetics of PF11 guttate pills were systematically studied.The following innovative results were obtained:1.Screen the anti-myocardial ischemia activity of PF11 Pills for the first time.The therapeutic and preventive anti-myocardial ischaemic effect of PF11 was evaluated in model rats administrated with PF11 pills(3,6,12 mg/kg;ig).Results showed that the left ventricular diameter(LVIDs)and volume(LVVs)were reduced,the left ventricular posterior wall thickness(IVSs),ejection fraction(EF%),fractional shortening(FS%)and heart contractility were increased significantly at the doses of 6 and 12 mg/kg.The levels of creatine kinase(CK),lactate dehydrogenase(LDH),aspartate transaminase(AST),malondialdehyde(MDA),and cardiac troponin T(cTnT)were also decreased accompanied with the activity of superoxide dismutase(SOD)increased.Meanwhile,decreasing infarct size and improving cardiomyocyte characteristic morphology were also observed obviously.So,we can draw a conclusion that PF11,simililar to positive drug metoprolol,has a good anti-myocardial ischaemic effect.2.Discuss the underlying mechanism of anti-myocardial ischemia effect of PF11 for the first time.Molecular pharmacology and metabolomics technique were used to explore the underlying mechanism of anti-myocardial ischemia effect of PF11 about the protein expression and metabolomic profiling.Results indicated that PF11 exerted anti-myocardial ischemia effect either by blocking beta 1 adrenergic receptor(?1-AR)or by regulating linoleic acid metabolism,arachidonic acid metabolism and sphingolipid metabolism.Molecular pharmacology study showed that the ?1-AR increased expression caused by myocardial ischemia(MI)in H9c2 cell could be reduced significantly by pre-administration with PF11.So,it is suggested that PF11 could antagonize the agonistic effects of neurotransmitters and catecholamines on ?1-AR by binding with?1-AR.In the molecular docking study,stable hydrogen bonds were observed between PF11 and ASP-322,ASP-200,and ASN-329 residues.The good space and electrical complementarity provided the further validation for PF11 as a ?1-AR blocker.Nontargeted metabolomic analysis showed that PF11 could reverse the abnormal change of 16 endogenous metabolites caused by MI.So,linoleic acid metabolism,arachidonic acid metabolism and sphingolipid metabolism derived from endogenous metabolites were considered as the main regulatory pathways.3.Study the pharmacokinetics of rats oral administered PF11 Pills for the first time(1)A UPLC-MS/MS quantitative method for the simultaneous determination of PF11 and its metabolites(RT5 and Ocotillol)in biological samples was established.LLOQ,linear range,specificity,accuracy,precision,dilution reliability,extraction recovery,matrix effects,and stability were all meet the requirements of pharmacokinetic studies.(2)The mean plasma concentration-time profiles of PF11 were determined.Some parameters such as t1/2,AUC,CL,Vz and others were calculated by DAS 3.0 software.Results showed the low AUC and wide body distribution of prototype drug.PF11 was the principal form before Tmax(0.78 h).While both PF11 and RT5 were detected during the elimination period(0.78-12.78 h).During the period of 12-48 h,only ocotillol was detected in plasma.(3)The tissues distribution of PF11 in rats were illustrated.Results showed that PF11 and RT5 were distributed widely in stomach,large intestine,uterus,brain and other tissues.The concentrations of PF11 and RT5 in tissues were greater than in plasma.While Ocotillol,the main metabolite,was evenly distributed in tissues and plasma.In a word,there were some differences in every pharmacokinetic process of PF11?RT5 and Ocotillol.(4)The excretion of PF11 in rats was studied.Results showed that excretion rates of PF11 in feces and urine reached the maximum during the period of 8-12 h,and cumulative excretion amount of PF11 in feces and urine were 33.19%and 6.07%respectively within 72 h.The maximum excretion rates of RT5 in feces and urine were at the period of 8-12 h.The cumulative excretion amount of PF11 in feces and urine were 0.25%and 0.15%respectively within 72 h.While,the maximum excretion rates of Ocotillol in feces and urine were at the period of 12-24 h.The cumulative excretion amount of PF11 in feces and urine were 14.19%and 6.98%respectively within 72 h.Totally,the sum of cumulative excretion amount of PF11,RT5 and Ocotillol was 60.83%.(5)The metabolites of PF11 in rats were identified.Results showed that PF11 was widely metabolized in rats.A total of 29 metabolites,including 9 phase ? metabolites(mainly included RT5 and Ocotillol)and 20 phase ? metabolites(mainly were cysteine binding and glutathione binding products),were characterized.Phase ?metabolic pathway included dehydration,hydrogenation,hydroxylation,.While Phase? metabolic pathway,as the predominated pathway,included methylation,acetylation,sulfation,phosphorylationcysteine binding,glycine binding,glutathione binding,and glucuronidation.In summary,PF11 had the anti-myocardial ischemia effect,such as improving heart function,regulating myocardial enzyme activity,alleviating oxidative stress,decreasing infarct size and improving cardiomyocyte characteristic morphology.The underlying mechanism may be either blocking ?1-AR or regulating linoleic acid metabolism,arachidonic acid metabolism and sphingolipid metabolism.PF11 and its main metabolites RT5?Ocotillol were distributed widely and eliminated slowly in rats.They were apt to bind with the tissues,while AUC was relatively lower.PF11 was widely metabolized in feces and urine,and the total cumulative excretion amount of prototype drug PF11 and metabolite Ocotillol was more than 60%.This thesis will provide the theoretical basis and data support for the further research and development of PF11 Pill,the innovative anti-myocardial ischemia drug.Meanwhile,it could offer a reference of new ideas and approaches to the PK-PD research of the similar drug.