Structure-Bioactivity Relationship for Benzimidazole Thiophene Inhibitors of Polo-Like Kinase 1 (PLK1), a Potential Drug Target in Schistosoma mansoni.
Structure-Bioactivity Relationship for Benzimidazole Thiophene Inhibitors of Polo-Like Kinase 1 (PLK1), a Potential Drug Target in Schistosoma mansoni.
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DOI：
10.1371/journal.pntd.0004356
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发表时间：
2016-01
影响因子：
3.8
通讯作者：
Caffrey CR
中科院分区：
文献类型：
--
作者：
Long T;Neitz RJ;Beasley R;Kalyanaraman C;Suzuki BM;Jacobson MP;Dissous C;McKerrow JH;Drewry DH;Zuercher WJ;Singh R;Caffrey CR
血吸虫扁虫寄生虫会导致血吸虫病，这是发展中国家的一种慢性贫困疾病。吡喹酮用于治疗和疾病控制。然而，它的功效谱是不完整的（对寄生虫的未成熟阶段活性较低或无活性），并且存在耐药性的担忧。因此，需要确定新的药物和药物靶标。我们发现，RNA干扰（RNAi）的曼氏血吸虫人类polo样激酶（huPLK）1直系同源物elevant一个有害的表型改变感染后的幼虫（童虫或童虫）。幼鱼和成虫的表型筛选与分析。mansoni与huPLK 1的小分子抑制剂一起鉴定了许多有效的抗肿瘤药物。其中有一种葛兰素史克（GSK）苯并咪唑噻吩抑制剂，已完成治疗实体瘤恶性肿瘤的I期临床试验。然后，我们获得了GSK公布的激酶抑制剂集（PKIS）1和2，并在表型上筛选了38种苯并咪唑噻吩PLK 1抑制剂的扩展系列。计算分析的控制和PLK 1信使处理群体的somule表现出独特的表型分布。利用主成分分析（PCA），这些群体表现出的表型进行了映射，可视化和分析，通过投影到一个低维空间。表型分布被发现有一个独特的形状和拓扑结构，这可以引起使用聚类分析。一个结构活性关系（SAR）被确定为苯并咪唑噻吩，保持为童虫和成虫寄生虫。最有效的抑制剂在1-2 μM浓度下1 h内产生显著的表型改变。其中包括先前在细胞测定中表征为huPLK 1的有效抑制剂的化合物。反向遗传和化学SAR数据支持继续调查的SmPLK 1作为一个可能的药物靶点和/或起诉的苯并咪唑噻吩化学型作为一种新的抗肿瘤药物的来源。只有一种药物可用于治疗血吸虫病，这是一种影响发展中国家数亿人的寄生虫病。因此，在寻找新药和药物靶点的过程中，我们对血吸虫版本的人马球样激酶（huPLK）1感兴趣，huPLK 1是一种在细胞分裂中具有关键功能的酶。我们使用RNA干扰来敲低SmPLK 1的信使RNA-曼氏血吸虫寄生虫的huPLK 1版本。这种干扰引起了破坏性的变化，在形态学的不成熟的“somule”阶段的寄生虫，这表明SmPLK 1是一个重要的蛋白生存。然后，我们从葛兰素史克（GSK）购买或获得了huPLK 1的各种小的化学抑制剂，并在培养物中测试了这些抑制剂对童虫和成虫寄生虫的作用。这些抑制剂中的许多引起了寄生虫的严重变化，并且对于somule，可以通过计算映射这些差异并将其与未暴露的寄生虫区分开来。对于GSK抑制剂，我们观察到“somule-adult生物活性聚类”，也就是说，对成虫有活性的化学物质对somule也有活性。这表明抑制剂中的某些化学属性受到青睐。有趣的是，许多对寄生虫最有效的GSK抑制剂也被认为可以有效地抑制huPLK 1并杀死癌细胞。总的来说，我们的数据表明SmPLK 1是一个可能的药物靶点，GSK的化学物质可以成为开发治疗血吸虫病新药的基础。
Schistosoma flatworm parasites cause schistosomiasis, a chronic and debilitating disease of poverty in developing countries. Praziquantel is employed for treatment and disease control. However, its efficacy spectrum is incomplete (less active or inactive against immature stages of the parasite) and there is a concern of drug resistance. Thus, there is a need to identify new drugs and drug targets. We show that RNA interference (RNAi) of the Schistosoma mansoni ortholog of human polo-like kinase (huPLK)1 elicits a deleterious phenotypic alteration in post-infective larvae (schistosomula or somules). Phenotypic screening and analysis of schistosomula and adult S. mansoni with small molecule inhibitors of huPLK1 identified a number of potent anti-schistosomals. Among these was a GlaxoSmithKline (GSK) benzimidazole thiophene inhibitor that has completed Phase I clinical trials for treatment of solid tumor malignancies. We then obtained GSKs Published Kinase Inhibitor Sets (PKIS) 1 and 2, and phenotypically screened an expanded series of 38 benzimidazole thiophene PLK1 inhibitors. Computational analysis of controls and PLK1 inhibitor-treated populations of somules demonstrated a distinctive phenotype distribution. Using principal component analysis (PCA), the phenotypes exhibited by these populations were mapped, visualized and analyzed through projection to a low-dimensional space. The phenotype distribution was found to have a distinct shape and topology, which could be elicited using cluster analysis. A structure-activity relationship (SAR) was identified for the benzimidazole thiophenes that held for both somules and adult parasites. The most potent inhibitors produced marked phenotypic alterations at 1–2 μM within 1 h. Among these were compounds previously characterized as potent inhibitors of huPLK1 in cell assays. The reverse genetic and chemical SAR data support a continued investigation of SmPLK1 as a possible drug target and/or the prosecution of the benzimidazole thiophene chemotype as a source of novel anti-schistosomals. Just one drug is available to treat schistosomiasis, a parasitic disease that affects hundreds of millions of people in developing countries. In the search for new drugs and drug targets, therefore, we have been interested in the schistosome version of human polo-like kinase (huPLK)1, an enzyme with critical functions in cell division. We used RNA interference to knock down messenger RNA for the SmPLK1 –the Schistosoma mansoni parasite’s version of huPLK1. This interference caused disruptive changes in the morphology of the immature ‘somule’ stage of the parasite, indicating that SmPLK1 is an important protein for survival. We then purchased, or acquired from GlaxoSmithKline (GSK), various small chemical inhibitors of huPLK1 and tested these against both the somules and adult parasites in culture. Many of these inhibitors caused severe changes in the parasite and, for somules, the differences could be computationally mapped and distinguished from unexposed parasites. For the GSK inhibitors, we observed ‘somule-adult bioactivity clustering,’ that is, chemicals active against the adults were also active against somules. This suggests that certain chemical attributes in the inhibitors are being favoured. Interestingly, many of the GSK inhibitors most active against the parasite are also known to both potently inhibit huPLK1 and kill cancer cells. Overall, our data suggest that SmPLK1 is a possible drug target and that the GSK chemistries could form the basis for developing a new drug to treat schistosomiasis.