In Vitro And In Vivo Study On Antidermatophytes Activity Of TTS-12,a Kind Of Steroid Saponin,and It's Preparation

Objective:Study antidermatophytes activity of TTS-12 by several indexes.Explore antidermatophytes mechanism of TTS-12 preliminarily.Model drug-resistantdermatophytes by in vitro antifungal drugs induced to study in vitro antifungal activity of TTS-12 against drug-resistant dermatophytes.Study antifungal activity of TTS-12against drug-resistant dermatophytes by a model of guinea pig infectedFCZconazole-resistant T.rubrum.Methods:1.Minimal inhibitory concentration?MIC?and minimal fungicide onconcentration?MFC?of TTS-12 against 2 kinds of common dermatophytes were texted by M38-A2 proposed by CLSI.The effect of hypa growth,spore germination andmycelial weight by TTS-12 were also researched.Checkerboard assay was adopted to investigate effect of drug combination use.2.Ergosterol content in cytomembrane of T.rubrum under different concentrations of TTS-12 were measured by HPLC-UV.Activity of two rate-limiting enzymes inmitochondria under TTS-12 also measured.3.Model FCZconazole-resistant and KCZoconazole-resistant 2 kinds of T.rubrum by in vitro increasing concentration method.Investigate the stability of two model and explore their sensibility to TTS-12.4.Set animal model of gunea pig infected FCZconazole-resistant T.rubrum.After administration of 1%TTS-12 gels,observe the effect of infection improvement.Results:1.For T.tumbrum,MIC of TTS-12 against standard strain is 2?g×mL^-1,and MFC is16?g×mL^-1,MIC range against 27 clinical isolated strains is 1⁸?g×mL^-1.ForT.mentagrophytes,MIC of TTS-12 against standard strain is 1?g×mL^-1,and MFC is8?g×mL^-1,MIC range against 27 clinical isolated strains is 0.5⁸?g×mL^-1.Concentration-dependent inhibitaion effect of hypa growth,spore germination and mycelial weight were shown under TTS-12.For trichophyton rubrum,using TTS-12with terbinafine hydrochloride cross combination,the proportion of FIC?0.5 reached81.5%.For trichophyton rubrum,using TTS-12 with terbinafine hydrochloride cross combination,the proportion of FIC?0.5 reached 76.2%.2.Setting up measurement of ergosterol by HPLC-UV.Higher concentration of TTS-12 can suppress ergosterol synethesis more effective.TTS-12 also can suppress activity of malic dehydrogenase and succinate dehydrogenase in aconcentration-dependent way.3.Setting up FCZconazole-resistant and KCZoconazole-resistant 2 kinds ofT.rubrum model.MIC ranges of two drugs are 128⁵12?g×mL^-1 and 64¹28?g×mL^-1.After several passage,the stability were kept.MIC of TTS-12 against 2 models were still in low level and ranges are are 4¹6?g×mL^-1 and4⁸?g×mL^-1.4 Animal model of gunea pig infected FCZconazole-resistant T.rubrum has been set up.Compared with model group,1%KCZoconazole and TTS-12 gels can improve infections,effect of TTS-12 was better.HE and PAS dyeing biopsy showed thatTTS-12 group inflammatory cells infiltration was light,cuticle was normal and stratum spinosum was not hypertrophy.