Yıldız Technical University, Faculty of Chemical and Metallurgical Engineering, Department of Bioengineering, Davutpasa campus, 34210, Istanbul, Turkey; Artvin Coruh University, Vocational School of Health Services, Medical Laboratory Techniques, 08000, Artvin, Turkey.
İstanbul Yeni Yüzyıl University, Faculty of Pharmacy, Department of Pharmaceutical Biotechnology, 34010, Istanbul, Turkey.
Bioorg Chem. 2021 May;110:104796. doi: 10.1016/j.bioorg.2021.104796. Epub 2021 Mar 5.
Tropical theileriosis is among the most common vector-borne diseases and caused by Theileria parasites. Theileria annulata is an obligate intracellular protozoan parasite and transmitted to especially Bos taurus and Bos indicus by Hyalomma tick vectors. C8 ([4-(3,4-dimethoxyphenyl)-6,7-dihydroxy-2H-chromen-2-one); C9 (4-(3,4-dihydroxyphenyl)-7,8 dihydroxy-2H-chromen-2-one); C21 (4-(3,4-dihydroxyphenyl)-6,7-dihydroxy-2H-chromen-2 one) were identified as potent Theileria annulata enolase (TaEno) inhibitors in our previous studies. An ideal drug compound must inhibit the target parasite enzyme without inhibiting its homolog in the host. In this study, the inhibitory effect of the compounds previously evaluated on TaEno were tested on the host Bos taurus enolase (BtEno3) by in vitro studies. The interactions of enzyme-coumarin and enzyme-coumarin-substrate by in silico studies were also performed. All of the coumarin derivatives tested showed very low inhibitory effects on B. taurus enolase; 36,87% inhibition at 100 μM concentration for C8, 8,13% inhibition at 100 μM concentration for C9 and 77,69 μM of IC value for C21. In addition, these three coumarin derivatives and substrate 2PG were docked into the BtEno3 using molecular docking methods. Molecular interactions between enolase-coumarin and enolase-coumarin-substrate complexes were analyzed using molecular dynamics simulation methods for 100 ns. Estimated free energy of bindings of the substrate 2PG and coumarin derivatives to the BtEno3 were calculated by MM-GB(PB)SA methods. In comparison to the inhibition studies performed on TaEno, C8 and C9 coumarin derivatives remain the possible inhibitor candidates as they inhibit the host enolase at very high concentrations. These two promising compounds will be further analyzed by in vitro and in vivo studies towards developing an alternative drug against tropical theileriosis.
热带泰勒虫病是最常见的虫媒传染病之一，由泰勒虫寄生虫引起。环形泰勒虫是一种专性细胞内原生动物寄生虫，通过璃眼蜱属的蜱虫传播给牛科动物，尤其是黄牛和水牛。在我们之前的研究中，C8（4-(3,4-二甲氧基苯基)-6,7-二羟基-2H-色烯-2-酮）；C9（4-(3,4-二羟基苯基)-7,8-二羟基-2H-色烯-2-酮）；C21（4-(3,4-二羟基苯基)-6,7-二羟基-2H-色烯-2-酮）被鉴定为潜在的环形泰勒虫烯醇酶（TaEno）抑制剂。一种理想的药物化合物必须抑制靶寄生虫酶而不抑制宿主中的同源物。在这项研究中，通过体外研究测试了之前评估的化合物对宿主水牛烯醇酶（BtEno3）的抑制作用。还通过计算机模拟研究进行了酶-香豆素和酶-香豆素-底物的相互作用。测试的所有香豆素衍生物对牛烯醇酶的抑制作用都非常低；在 100 μM 浓度下，C8 的抑制率为 36.87%，C9 的抑制率为 8.13%，C21 的 IC 值为 77.69 μM。此外，这三种香豆素衍生物和底物 2PG 被使用分子对接方法对接进 BtEno3 中。使用分子动力学模拟方法对 100 ns 时间内烯醇酶-香豆素和烯醇酶-香豆素-底物复合物之间的分子相互作用进行了分析。通过 MM-GB(PB)SA 方法计算了底物 2PG 和香豆素衍生物与 BtEno3 的结合自由能。与在 TaEno 上进行的抑制研究相比，C8 和 C9 香豆素衍生物仍然是可能的抑制剂候选物，因为它们在非常高的浓度下抑制宿主烯醇酶。这两种有前途的化合物将通过体外和体内研究进一步分析，以开发针对热带泰勒虫病的替代药物。
