目的:合成碳酸酐酶的新型磺酰胺抑制剂,并开发体外优先工作流程以选择用于体内评估的化合物。
背景:碳酸酐酶(CA)抑制剂在青光眼药物发现研究的背景下获得了极大的关注,缺氧性恶性肿瘤和细菌感染。在以前的作品中,我们已经成功地使用直接磺氯化方法来开发不同的杂环伯磺胺类具有显着的活性和选择性对治疗相关的CA亚型。
目的:新型三氟甲基异恶唑基-和三氟甲基吡唑基-取代的(杂)芳族磺酰胺的合成和CA抑制性能的研究。
方法:通过羟基异恶唑啉和吡唑的直接磺氯化,然后与氨反应,合成了13种三氟甲基异恶唑基和13种三氟甲基吡唑基取代的(杂)芳族磺酰胺。通过1H和13CNMR以及元素分析证实了化合物的结构。对得到的化合物进行评价,使用CA酯酶活性测定,因为它们有可能阻断牛CA(bCA)的催化活性。
结果:使用热转移测定(TSA)测试了基于酯酶活性测定数据选择的八种最有效的化合物对酶的直接亲和力。这些化合物显示的Kd值(通过TSA测量)在两位数纳摩尔范围内,因此显示出与参比药物乙酰唑胺相当的活性。
结论:将bCA酯酶活性测定与热转移测定相结合代表了一种简化且经济的策略,用于优先考虑磺胺CA抑制剂,以进行后续体内评估。
OBJECTIVE: To synthesize novel sulfonamide inhibitors of carbonic anhydrase and develop in vitro prioritization workflow to select compounds for in vivo evaluation.
BACKGROUND: Carbonic anhydrase (CA) inhibitors gain significant attention in the context of drug discovery research for glaucoma, hypoxic malignancies, and bacterial infections. In previous works, we have successfully used direct sulfochlorination approach to develop diverse heterocyclic primary sulfonamides with remarkable activity and selectivity against therapeutically relevant CA isoforms.
OBJECTIVE: Synthesis and investigation of the CA inhibitory properties of novel trifluoromethylisoxazolyl- and trifluoromethylpyrazolyl-substituted (hetero)aromatic sulfonamides.
METHODS: Thirteen trifluoromethylisoxazolyl- and thirteen trifluoromethylpyrazolyl-substituted (hetero) aromatic sulfonamides were synthesized by direct sulfochlorination of hydroxyisoxazolines and pyrazoles followed by reaction with ammonia. The compound structures were confirmed by 1H and 13C NMR as well as element analysis. The obtained compounds were evaluated, using the CA esterase activity assay, for their potential to block the catalytic activity of bovine CA (bCA).
RESULTS: Eight most potent compounds selected based on the esterase activity assay data were tested for direct affinity to the enzyme using the thermal shift assay (TSA). These compounds displayed Kd values (measured by TSA) in the double-digit nanomolar range, thus showing comparable activity to the reference drug acetazolamide.
CONCLUSIONS: Coupling the bCA esterase activity assay with thermal shift assay represents a streamlined and economical strategy for the prioritization of sulfonamide CA inhibitors for subsequent evaluation in vivo.