The Design, Synthesis Of New Difunctional Nitric Oxide Donor Drugs And Their Initial Pharmacodynamic Study

Cardiovascular and cerebrovascular diseases have increasing morbidity due to thechanges in dietary habits and lifestyles of the population which are caused by thedevelopment of the society. Ischemic heart disease (IHD), accompany with heart failure asits main cause of death, is a common disease which seriously endager human health. Theeffective measure to save acute myocardial ischemia is the recovery of blood flow incoronary at time, while the recovery of blood flow will lead to myocardial ischemia/reperfusion injury (MIRI). How to effectively mitigate the myocardial damage has becomethe biggest challenge in the treatment of IHD.Pulmonary arterial hypertension (PAH) is also one kind of common clinical diseasewhich is difficult to be cured. The increase of pulmonary arterial resistance and highpressure are the clinical features of PAH. The pathological features of PAH include thepulmonary artery vasospasm, intimal hyperplasia and remodeling, which can lead to aprogressive increase in pulmonary resistance and eventually associate with rightventricular hypertrophy and end-stage heart failure. Currently, the treatment of PAH is stilllack of effective methods. PAH has high mortality and morbidity and poor prognosis. It’sso difficult to be cured that there’s still no effective drug so far. The researches prove that the pathological processes of MI/R and PAH both associatewith the free radical damage and the synthesis reduction of nitric oxide (NO). Isosorbidemononitrate (IM) is a kind of NO donor drug which can treat both MIRI and PAH.However, it hasn’t the antioxidant effect and is easy to produce drug resistance afterlong-term using. Natural medicine ferulic acid and resveratrol are free radical scavengers,they both have the certain effect in treatment of MIRI and PAH.Recently years, people have begun to combine the structure of NO donor with knowndrug molecules in order to increase the activity of drugs. Therefore, the aim of this study isto make ferulic acid and resveratrol as lead compounds to combine with IM respectively,and get two kinds of difunctional drugs AcFA-201and BZ-1. Their pharmacodynamicresearches have also been made.There are two parts of experiments have been made in this study:Part one:1. The synthesis of AcFA-201At first transfered the hydroxyl of IM into amino, and combined it with acetyl ferulicacid, then we got the new compound AcFA-201which own a stable structure. Thesynthesis methods of AcFA-201are optimized. To confirm the structure of AcFA-201, theMS, IR,1HNMR have been tested. The physicochemical properties of AcFA-201are alsotested.2. The research of protective effect of AcFA-201on ischemic myocardium ratsMale Sprague-Dawley rats, subjected to30minutes of myocardial ischemia and3hours of reperfusion. Then the the48rats were randomly assigned into6groups. They arecontrol (SHAM) group, model (MI/R) group, sodium ferulate (SF) group, isosorbidemononitrate (IM) group, AFI group and AcFA-201group. The doses of SF and IM are40mg/kg and the doses of AFI and AcFA-201are10mg/kg before surgery. After theexperiment, we detected three parts of indicators include heart function, the content of NO,H2O2, MDA and the activity of CK, SOD and LDH.The results showed that LVDP,±dp/dtmax, the content of NO and the activity of SODin serum are reduced significantly (P<0.05), and the content of H2O2, MDA and the activity of CK, LDH in serum are improved markedly (P<0.05) in model group comparedwith control group. All indicators are significantly ameliorated in four drugs treatmentgroups compared with model group (P<0.05or P<0.01). AcFA-201group and AFI groupimproved the heart function (P<0.05), reduced the activity of CK, LDH and content ofH2O2, MDA and improved the activity of SOD and content of NO in serum moresignificant than SF and IM group (P<0.05or P<0.01). AFI group showed the similar trendwith AcFA-201group (P0.05).3. The research of stability in gastrointestinal fluid of AcFA-201and AFIThe vitro incubation method was used to research the stability of AcFA-201and AFI ingastrointestinal fluid. The results showed that the amino bond is more stable than the esterbond, and AcFA-201is more suitable to be developed into oral drug.Part two:1. The synthesis of compound BZ-1At first we got the intermediate compound methylated resveratrol, and combined itwith IM, then we got the new compound BZ-1. The research of the synthesis methodsshowed that condensation method is the most avalible method. To confirm the structure ofBZ-1, the MS, IR,1HNMR have been tested. The physicochemical properties of BZ-1arealso tested.2. The NO release ability in vivo of BZ-1The NO release ability in vivo of BZ-1has been studied with IM as control drug. Thedatas showed that BZ-1has an ability of release NO in vivo, as which is weaker than IMtotally.3. The effect of BZ-1on hypoxia in miceThe effect of BZ-1on hypoxia in mice has been tested with IM as control drug. Theresult showed that the high dose group of BZ-1can markedly improve the survival time ofmice, while IM didn’t show this effect.4. The effect of BZ-1on MCT-induced pulmonary arterial hypertension in ratsThe method of one-time injection monocrotaline (MCT) was used to establish theanimal model and randomly assigned the50male Sprague-Dawley rats into5groups. They are control group, model group, low dose of BZ-1group, middle dose of BZ-1group,high dose of BZ-1group. The doses of BZ-1are5,15,25mg/kg. The solvent given to thecontrol group and model group is twain80: saline (1:40, v: v). After the experiment, wedetected several indicators include mean pulmonary arterial pressure (mPAP) and rightventricle hypertrophy index (RVHI), whole blood viscosity (WBV) and erythrocyteaggregation index (EAI), the NO content and NOS activity in the serum and lung and andpathological section observation.The results showed that the mPAP, RVHI, WBV and EAI of model group areimproved significantly compared with control group (P<0.05). The NO content and NOSactivity in the serum and lung of model group are markedly reduced, the vessel wallthickness/total vascular diameter (WT%) and vessel wall area/total vascular area (WA%)are markedly increased compared with control group (P<0.05). The middle dose groupand high dose group of BZ-1can reduce the mPAP, RVHI, WBV, EAI, improved the NOcontent and NOS activity in the serum and lung significantly compared with the modelgroup (P<0.05or P<0.01). The observation datas of WT%and WA%are also beenmarkedly decreased in middle dose group and high dose group (P<0.05or P<0.01). Lowdose group didn’t show the significance (P0.05).This study designed and synthesized two kinds of new compounds AcFA-201andBZ-1. The synthesis methods are optimized and the structures of the two compounds areconfirmed respectively. The pharmacodynamic researches of the two compounds have alsobeen made. The research of protective effect of AcFA-201on ischemic myocardium ratsshowed that AcFA-201can effectively improve ischemia/reperfusion heart function andreduce myocardial injury. The myocardial protection mechanism may be related to theraise of the content of NO, the dilation of blood vessels, the decrease ofischemia/reperfusion injury. The amide bond structure made it more suitable for thedevelopment of oral drugs. The effect of BZ-1on MCT-induced pulmonary arterialhypertension in rats indicated that BZ-1is effective in improving pulmonaryvasoconstriction, improving NO content, NOS activity and hemorrheology indexes, andreducing pulmonary arterial hypertension in rats. BZ-1significantly reduced pulmonary vascular wall thickening and luminal stenosis, thereby prohibited the pathological lungtissue remodeling. BZ-1can also release NO in vivo and improve the survival time ofmice. It’s hoped to become a novel drug which can treat PAH.