XO、 ACE、FXa Inhibitor And Antioxidant Activity In Vitro Screening And Establishment Of KPC-108Model Of Rats With Adriamycin Nephropathy

Objective: Establishment of xanthine oxidase (XO), angiotensin convertingenzyme (ACE), blood coagulation factor Xa (F Xa) method of screening inhibitor andantioxidant activity in vitro of Institute, KPC samples were in vitro screened; Basedon the analysis of screening results, we selected samples which have potential clinicalprospect to investigate the effectiveness with in vivo nephropathy model and learnsafety study, so as to provide experimental basis for further research and development.Mothed:Using the DPPH, OH-, O2-, ABTS scavenging capacity assay, TOCdetermination, determination of oxidation activity inhibited the adjacent benzene threephenolic antioxidant activity to investigate the Antioxidant activity of samples; Usingthe determination of kinetic parameters of the enzyme reaction system byspectrophotometry to study the tested samples effects on ACE, XOD, F Xaactivity;Using the adriamycin nephropathy rats model to investigate the influence of2kinds of compounds for animal model of urinary protein, blood biochemistry, bloodflow dynamics; The adriamycin nephropathy rats model was once more used toinvestigate the effectiveness in screening from the urinary protein, blood biochemistryand antioxidant activity, hemodynamics, renal tissue morphology by the experiment.And the maximum tolerance of the preliminary was used to investigate its safety.Results:1. From the26candidate samples screened, KPC-021, KPC-108,KPC-056, KPC-002, KPC-070compounds have strong antioxidant activity; thesample KPC-108, KPC-056can inhibit angiotensin converting enzyme activity;KPC-010, KPC-108had strong inhibitory effects on the activity of XOD; Combinedwith compound structure, activity and existing research results, we select KPC-056KPC-108, two samples with vascular lesions, increasing incidence, clinical necessary nephropathy model rats to be treated with medication observation.2.KPC-108KPC-056, two samples with20mg/kg dosage by intragastricadministration for4weeks, they can significantly reduce Cre, NO levels in animalmodels serum, have the trend of reduce the increased proteinuria; Can decrease BUN,TG and whole blood viscosity in different degree, the rise of TP and ALB; KPC-108can significantly reduce the level of MDA. Compared with KPC-056, KPC-108reduced the levels of serum Cre, TG, MDA and whole blood viscosity is the mostobvious, compared with the model group were significant differences. So we chosesample KPC-108for a second round of in vivo dose of pharmacodynamics andpreliminary safety validation study.3. KPC-108to20,40mg/kg dosage by intragastric administration for8weeks,can significantly reduce the model animal protein urine, hypercoagulable statecorrection of hypoproteinemia, hyperlipemia and blood, reducing serum lipidperoxide content in rat models, the expression of TNF-α inhibits renal tissues, andhas a protective effect on renal tissue, the improvement in renal function. Samples ofKPC-108mice were intragastrically administered with maximum tolerance dose≥18g/kg, there are no obviously acute toxic reaction and death.Conclusions: To investigate the protective effects of KPC-108on adriamycin nephritismodel of rat kidney injury, the mechanism may be as follows:(1)By reducing urinaryprotein, blood fat, reduce two of direct renal damage;(2) Due to the structure andfunction of glomerular filtration membrane lipid peroxidation reaction of glomerularepithelial cells from the damage reduced by antioxidant effect;(3)Inhibited theinfiltration of mononuclear macrophage in kidney and plasma to reduce levels of TNF-α;(4) Improve renal blood perfusion, thereby improving the renal pathologicalchanges.