Thirteen pairs of I3P enantiomers were screened using nine polysaccharide chiral stationary phases and three different mobile phases. The purification strategy for 13 pairs of I3P enantiomers were designed and optimized considering enantiomeric purity and enrichment of isomers. Out of 13 I3P derivatives which were screened using supercritical fluid chromatography, 10 derivatives displayed excellent baseline separation using a Lux Cellulose—4 column and their resolution from higher to lower order of I3P-11, 13, 4, 12, 2, 1, 9, 3, 7 and 8 derivatives whereas in case of Lux Cellulose—2 column, the moderate separation was achieved as compared to Cellulose—4 in the order I3P-5, 6 and 10 derivatives. Excellent enantiomeric separations and retentions for all 13 I3P enantiomer derivatives were obtained in Cellulose—4 and Cellulose—2 columns in presences of methanol as organic modifier without any additives except in the case of I3P 12 enantiomer. The absolute stereochemical assignment of the purified isomers was determined through an optical rotation study. Among the series of I3P derivatives, I3P-5 showed potent antioxidant activity against catalase with an IC50 value of 13.78 μM. Further molecular docking, MM/GBSA and molecular dynamics studies revealed that the I3P-5 derivatives effectively bind to catalase with a docking score of −5.41 kcal/mol. Which validated chiral docking and indicated great potential for enantiomeric separation in drug discovery and present studies (R)—enantiomer preferentially depicts good binding capacity with catalase.

中文翻译：

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多糖基手性固定相超临界流体色谱法分离吲哚-3-丙酰胺衍生物的对映异构体

使用九种多糖手性固定相和三种不同的流动相筛选了 13 对 I3P 对映异构体。考虑对映体纯度和异构体富集度，设计并优化了 13 对 I3P 对映体的纯化策略。在使用超临界流体色谱法筛选的 13 种 I3P 衍生物中，有 10 种衍生物使用 Lux Cellulose-4 色谱柱显示出出色的基线分离，并且它们的分离度从高到低顺序为 I3P-11、13、4、12、2、1、9 、3、7 和 8 衍生物，而在 Lux Cellulose-2 色谱柱的情况下，与 Cellulose-4 相比，以 I3P-5、6 和 10 衍生物的顺序实现了适度的分离。除 I3P 12 对映异构体外，所有 13 种 I3P 对映异构体衍生物均在 Cellulose-4 和 Cellulose-2 色谱柱中获得了出色的对映异构体分离和保留，其中甲醇作为有机改性剂，没有任何添加剂。纯化异构体的绝对立体化学归属通过旋光研究确定。在I3P系列衍生物中，I3P-5对过氧化氢酶具有很强的抗氧化活性，IC50值为13.78 μM。进一步的分子对接、MM/GBSA 和分子动力学研究表明，I3P-5 衍生物有效地与过氧化氢酶结合，对接分数为 -5.41 kcal/mol。