Activity In Vivo of Anti-Trypanosoma cruzi Compounds Selected from a High Throughput Screening
Activity In Vivo of Anti-Trypanosoma cruzi Compounds Selected from a High Throughput Screening
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DOI：
10.1371/journal.pntd.0001298
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发表时间：
2011-08-01
影响因子：
3.8
通讯作者：
Rodriguez, Ana
中科院分区：
文献类型：
--
作者：
Andriani, Grasiella;Chessler, Anne-Danielle C.;Rodriguez, Ana
包括重组病原体和快速检测方法在内的新技术正在促进治疗被忽视疾病的药物的开发。最近，报道了首次高通量筛选（HTS）的结果，以测试化合物对抗克氏锥虫锥鞭毛体感染宿主细胞的活性。我们从该 HTS 的命中结果中筛选出 23 种化合物，据报道这些化合物具有高抗锥虫活性和对宿主细胞低毒性。这些化合物经过高度纯化，其结构通过 HPLC/质谱法确认。这些化合物在体外进行了测试，其中大约一半证实了抗 T 蛋白。 HTS 中报告了 cruzi 活性，IC50 值低于 5 muM。我们还采用了快速测定法来测试抗 T。使用感染表达荧光素酶的转基因 T. cruzi 的小鼠作为急性感染模型，在体内合成 cruzi 化合物。使用该测定法还对体外具有活性的化合物进行了体内测试，我们发现两种具有相似结构和较低体外 IC50 值（0.11 和 0.07 μM）的相关化合物，在治疗五天后可将小鼠模型中的克氏锥虫感染减少 90% 以上。我们的研究结果证明了 HTS 等新技术对于被忽视疾病的药物发现途径的益处，但也警告需要在体外确认结果。我们还展示了基于表达荧光素酶的寄生虫的体内快速筛选方法对于在开发链早期确定化合物的优先顺序非常有用。
Novel technologies that include recombinant pathogens and rapid detection methods are contributing to the development of drugs for neglected diseases. Recently, the results from the first high throughput screening (HTS) to test compounds for activity against Trypanosoma cruzi trypomastigote infection of host cells were reported. We have selected 23 compounds from the hits of this HTS, which were reported to have high anti-trypanosomal activity and low toxicity to host cells. These compounds were highly purified and their structures confirmed by HPLC/mass spectrometry. The compounds were tested in vitro, where about half of them confirmed the anti-T. cruzi activity reported in the HTS, with IC50 values lower than 5 mu M. We have also adapted a rapid assay to test anti-T. cruzi compounds in vivo using mice infected with transgenic T. cruzi expressing luciferase as a model for acute infection. The compounds that were active in vitro were also tested in vivo using this assay, where we found two related compounds with a similar structure and low in vitro IC50 values (0.11 and 0.07 mu M) that reduce T. cruzi infection in the mouse model more than 90% after five days of treatment. Our findings evidence the benefits of novel technologies, such as HTS, for the drug discovery pathway of neglected diseases, but also caution about the need to confirm the results in vitro. We also show how rapid methods of in vivo screening based in luciferase-expressing parasites can be very useful to prioritize compounds early in the chain of development.