Abstract

Elevated plasma levels of hyaluronic acid (HA) is a disease marker in liver pathology and other inflammatory disorders. Inhibition of HA synthesis with coumarin 4-methylumbelliferone (4MU) has a beneficial effect in animal models of fibrosis, inflammation, cancer and metabolic syndrome. 4MU is an active compound of approved choleretic drug hymecromone with low bioavailability and a broad spectrum of action. New, more specific and efficient inhibitors of hyaluronan synthases (HAS) are required. We have tested several newly synthesized coumarin compounds and commercial chitin synthesis inhibitors to inhibit HA production in cell culture assay. Coumarin derivative compound VII (10′-methyl-6′-phenyl-3′H-spiro[piperidine-4,2′-pyrano[3,2-g]chromene]-4′,8′-dione) demonstrated inhibition of HA secretion by NIH3T3 cells with the half-maximal inhibitory concentration (IC50) = 1.69 ± 0.75 μΜ superior to 4MU (IC50 = 8.68 ± 1.6 μΜ). Inhibitors of chitin synthesis, etoxazole, buprofezin, triflumuron, reduced HA deposition with IC₅₀ of 4.21 ± 3.82 μΜ, 1.24 ± 0.87 μΜ and 1.48 ± 1.44 μΜ, respectively. Etoxazole reduced HA production and prevented collagen fibre formation in the CCl₄ liver fibrosis model in mice similar to 4MU. Bioinformatics analysis revealed homology between chitin synthases and HAS enzymes, particularly in the pore-forming domain, containing the proposed site for etoxazole binding.

中文翻译：

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新型香豆素化合物和几丁质合成抑制剂抑制透明质酸分泌

摘要

血浆透明质酸 (HA) 水平升高是肝脏病理学和其他炎症性疾病的疾病标志物。用香豆素 4-甲基伞形酮 (4MU) 抑制 HA 合成在纤维化、炎症、癌症和代谢综合征的动物模型中具有有益作用。4MU是一种获批利胆药物hymecromone的活性化合物，生物利用度低，作用广谱。需要新的、更特异和更有效的透明质酸合酶 (HAS) 抑制剂。我们已经测试了几种新合成的香豆素化合物和商业几丁质合成抑制剂，以抑制细胞培养试验中 HA 的产生。香豆素衍生物化合物VII（10'-甲基-6'-苯基-3'H-螺[哌啶-4,2'-吡喃并[3,2-g]色烯]-4'，8'-二酮)显示抑制 NIH3T3 细胞的 HA 分泌，其半数最大抑制浓度 (IC50) = 1.69 ± 0.75 μM，优于 4MU (IC50 = 8.68 ± 1.6 μM)。几丁质合成抑制剂、乙恶唑、噻嗪酮、氟嘧菌胺，通过 IC 减少 HA 沉积4.21 ± 3.82 μM、1.24 ± 0.87 μM 和 1.48 ± 1.44 μM 的 50 个_。在与 4MU 相似的小鼠的 CCl _{4肝纤维化模型中，乙恶唑降低了 HA 的产生并阻止了胶原纤维的形成。}生物信息学分析揭示了几丁质合酶和 HAS 酶之间的同源性，特别是在成孔结构域中，包含拟定的乙恶唑结合位点。