We have carried out a docking inspired synthesis and screening of a library of diazenyl-derivatives of pyridazinylpyrazolone molecules for their ability to modulate the amyloidogenic self-assembly of human islet amyloid polypeptide (hIAPP). hIAPP is a 37-residue peptide which is involved in glycemic control along with insulin. Its extracellular fibrillar assemblies in pancreatic β-cells are responsible for type 2 diabetes. A three-step synthetic scheme was used to prepare these novel compounds using 2-(6-chloropyridazin-3-yl)-5-methyl-2,4-dihydro-3H-pyrazol-3-one as a key intermediate that was reacted with various diazo electrophiles to generate a library of compounds with yields ranging from 64 to 85%. The effect of the compounds on hIAPP amyloid fibril formation was evaluated with a thioflavin T (ThT) fluorescence-based kinetic assay. Furthermore, TEM imaging was carried out to corroborate the interactions of the compounds with hIAPP and subsequent hIAPP inhibition at the different level of fibrillization. The CD spectroscopy showed that upon incubation with SSE15314 for 12 h, the percentage of α-helices was maintained to a level of hIAPP at 0 h. The current study presents identification and characterization of SSE15314 as the hit, which completely inhibited the fibril formation and can be further optimized into a lead compound.

中文翻译：

---

新型基于哒嗪基吡唑啉酮的重氮化合物的合成和鉴定，可作为人类胰岛淀粉样多肽聚集的抑制剂。

我们进行了对接启发性合成和筛选哒嗪基吡唑啉酮分子的二氮烯基衍生物的文库，因为它们具有调节人胰岛淀粉样多肽（hIAPP）的淀粉样生成自组装的能力。hIAPP是37个残基的肽，与胰岛素一起参与血糖控制。胰腺β细胞中的细胞外纤维状组件负责2型糖尿病。使用2-（6-氯哒嗪-3-基）-5-甲基-2,4-二氢-3H-吡唑-3-酮作为反应的关键中间体，采用三步合成方案制备了这些新型化合物。用各种重氮亲电试剂生成化合物库，收率范围为64％至85％。通过基于硫代黄素T（ThT）荧光的动力学分析评估了化合物对hIAPP淀粉样蛋白原纤维形成的影响。此外，进行TEM成像以证实化合物与hIAPP的相互作用以及随后的hIAPP在不同原纤维化水平的抑制。CD光谱表明，在与SSE15314一起温育12小时后，在0小时α-螺旋的百分率维持在hIAPP水平。当前的研究提出了对SSE15314的鉴定和表征，它完全抑制了原纤维的形成，并可以进一步优化为铅化合物。