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<i>Toxoplasma gondii</i>: A simple high-throughput assay for drug screening <i>in vitro</i>
Jin, ChunMei,Kaewintajuk, Kusuma,Jiang, JingHua,Jeong, WooJin,Kamata, Masaki,KIM, Hye-Sook,Wataya, Yusuke,Park, Hyun Elsevier 2009 Experimental parasitology Vol.121 No.2
<P><B>Abstract</B></P><P><I>Toxoplasma gondii</I> is the etiologic agent of toxoplasmosis. Although the combination of sulfadiazine and pyrimethamine is used as therapy for this disease, these drugs can have serious side effects and its use is limited in pregnancy. Therefore there is a need for new anti-<I>T. gondii</I> drugs in the clinic. Some systems for <I>T. gondii</I> drug screening have been described, but these have limitations and can be difficult. In order to solve these problems, we established a system to screen drugs <I>in vitro</I> that involved using cell viability methods to calculate drug selectivities, which are Trypan blue, [3-(4,5-dimethylthiazol-zyl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazoliuzolium, inner salt] (MTS) method and lactate dehydrogenase (LDH) assay. These assays were simple to establish and perform. The IC<SUB>50</SUB> values calculated from the morphological assay were not significantly different from the EC<SUB>50</SUB> values calculated using the other three methods. In particular, the results of the morphological assay showed a distinct association with the MTS assay (<I>R</I>=0.9841). These assays could be used for a wide range of applications in the screening of new drugs and may provide an alternative to the techniques currently used to screen for candidate anti-<I>T. gondii</I> compounds <I>in vitro</I>. In this study, we also tested many compounds and identified some that had a good anti-<I>T. gondii</I> effect <I>in vitro</I> based on the MTS assay. This simple and fast system allowed us to determine which compounds to investigate further using <I>in vivo</I> experiments.</P>
최화정,이재훈,여선주,Kusuma Kaewintajuk,이규양,김석,송현옥,박현 대한약학회 2015 Archives of Pharmacal Research Vol.38 No.5
Coccidiosis affects many vertebrates worldwide,but treatment with known anti-coccidial drugs causesseveral adverse side effects. There is a critical need for thedevelopment and evaluation of new drugs. The anti-coccidialeffect of 1-[4-(4-nitrophenoxy)phenyl]propane-1-one(NPPP), a synthetic compound, was studied in vitro andin vivo. Treatment with NPPP showed anti-Toxoplasmaactivity in vitro with a lower EC50 value than pyrimethamine. In ICR mice infected with Toxoplasma gondii, oraladministration of NPPP for 4 days showed statisticallysignificant anti-Toxoplasma activity with lower numbers oftachyzoite than those of the negative control (p\0.01). NPPP also exhibited strong anti-Eimeria activity in Eimeriatenella-infected chickens when treated for 4 days withorally administered NPPP at a dose of 100 mg/kg. Potentialtarget proteins of NPPP were analyzed by proteomic profilesof T. gondii tachyzoites. Two hypothetical proteinswere identified as possible targets of NPPP, a putativeortholog of vacuolar ATP synthase subunit C and a class IS-adenosylmethionine-dependent methyltransferase. Ourdata show that the NPPP might be an anti-coccidial drugcandidate for clinical application against coccidial infections. Future investigations will focus on identifying thefunction of proteins regulated by NPPP.