背景:应用基于GPCR(G蛋白偶联受体)靶标的方法来鉴定节肢动物特异性蜱激肽受体的拮抗剂。当通过RNA干扰使激肽受体沉默时,预计这些小分子会再现在微型动物和雌性动物中观察到的有害表型效应。微小根皮phalus,南方牛蜱,牛发烧蜱,或亚洲蓝蜱,是导致致命的牛babesiosis和无形体病的病原微生物的媒介。全球tick种群对杀螨剂的广泛抗性强调,探索有效控制tick的新目标势在必行。
结果:使用表达蜱激肽受体的CHO-K1细胞系,在“双重添加”钙荧光测定法中筛选了先前鉴定的蜱激肽拮抗剂的53种结构类似物。七个分子被验证为非细胞毒性拮抗剂,其中四个是部分的(SACC-0428764、SACC-0428780、SACC-0428800和SACC-0428803),三个是完全拮抗剂(SACC-0428799、SACC-0428801和SACC-0428815)。这些拮抗剂中的四种(SACC-0428764,SACC-0428780,SACC-0428799和SACC-0428815)也抑制了由肌力激肽激动剂类似物1728诱导的蜱中肠收缩,证实了它们的拮抗生物活性。在重组人神经激肽(NK)受体上测试了小分子,与无脊椎动物激肽受体最相似的一种。大多数分子是NK1受体的抑制剂,除了SACC-0412066,一种先前鉴定的蜱激肽受体拮抗剂,其仅在测试的最高浓度(25μm)下抑制NK1受体。没有一种分子抑制NK3人受体。
结论:通过这种方法鉴定的分子可能是研究蜱激肽信号系统和中肠生理学的有用探针。©2024作者(S)。害虫管理科学由JohnWiley&SonsLtd代表化学工业学会出版。
BACKGROUND: A GPCR (G protein-coupled receptor) target-based approach was applied to identify antagonists of the arthropod-specific tick kinin receptor. These small molecules were expected to reproduce the detrimental phenotypic effects that had been observed in Rhipicephalus microplus females when the kinin receptor was silenced by RNA interference. Rhipicephalus microplus, the southern cattle tick, cattle fever tick, or Asian blue tick, is the vector of pathogenic microorganisms causing the deadly bovine babesiosis and anaplasmosis. The widespread resistance to acaricides in tick populations worldwide emphasizes that exploring novel targets for effective tick control is imperative.
RESULTS: Fifty-three structural analogs of previously identified tick kinin antagonists were screened in a \'dual-addition\' calcium fluorescence assay using a CHO-K1 cell line expressing the tick kinin receptor. Seven molecules were validated as non-cytotoxic antagonists, four of which were partial (SACC-0428764, SACC-0428780, SACC-0428800, and SACC-0428803), and three were full antagonists (SACC-0428799, SACC-0428801, and SACC-0428815). Four of these antagonists (SACC-0428764, SACC-0428780, SACC-0428799, and SACC-0428815) also inhibited the tick midgut contractions induced by the myotropic kinin agonist analog 1728, verifying their antagonistic bioactivity. The small molecules were tested on recombinant human neurokinin (NK) receptors, the one most similar to the invertebrate kinin receptors. Most molecules were inhibitors of the NK1 receptor, except SACC-0412066, a previously identified tick kinin receptor antagonist, which inhibited the NK1 receptor only at the highest concentration tested (25 μm). None of the molecules inhibited the NK3 human receptor.
CONCLUSIONS: Molecules identified through this approach could be useful probes for studying the tick kinin signaling system and midgut physiology. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.