Discovery and Optimization of Indoline-Based Compounds as Dual 5-LOX/sEH Inhibitors: In Vitro and In Vivo Anti-Inflammatory Characterization

Abstract

The design of multitarget drugs represents a promising strategy in medicinal chemistry and seems particularly suitable for the discovery of anti-inflammatory drugs. Here, we describe the identification of an indoline-based compound inhibiting both 5-lipoxygenase (5-LOX) and soluble epoxide hydrolase (sEH). In silico analysis of an in-house library identified nine compounds as potential 5-LOX inhibitors. Enzymatic and cellular assays revealed the indoline derivative 43 as a notable 5-LOX inhibitor, guiding the design of new analogues. These compounds underwent extensive in vitro investigation revealing dual 5-LOX/sEH inhibitors, with 73 showing the most promising activity (IC 50 s of 0.41 ± 0.01 and 0.43 ± 0.10 μM for 5-LOX and sEH, respectively). When challenged in vivo in zymosaninduced peritonitis and experimental asthma in mice, compound 73 showed remarkable anti-inflammatory efficacy. These results pave the way for the rational design of 5-LOX/sEH dual inhibitors and for further investigation of their potential use as anti-inflammatory agents.

Design, Synthesis, and Pharmacological Characterization of a Potent Soluble Epoxide Hydrolase Inhibitor for the Treatment of Acute Pancreatitis

“…The conversion of urea to thiourea provides only one hydrogen bond with D335 ( 10 , Figure S66A). This observation agrees with our previous studies and with data reported in the literature, suggesting that the urea group is a crucial structural requirement to potently inhibit sEH . Moreover, it is here demonstrated that the amide group was not able to give the network of H-bonds observed for the urea group.…”

“…1 H NMR (400 MHz, CDCl 3 ): δ: 3.04 (t, 2H, CH 2 , J = 6.9 Hz); 4.30 (t, 2H, CH 2 , J = 6.9 Hz); 6. 54 (20). Intermediate 20 was synthesized starting from 5-nitroindole and methyl 4bromobutanoate following the procedure E. FC in dichloromethane/hexane 9/1, R f : 0.45.…”

“…19 We have recently described the pharmacological characterization of a dual 5-LOX/sEH inhibitor (compound 73, Figure 1) showing a remarkable potency in zymosan-induced peritonitis and in ovalbumin-induced asthma murine models. 20 These preliminary results led us to highlight the structural determinants leading to 5-LOX and sEH inhibition. In our previous paper, beyond the 5-LOX/sEH dual inhibitor 73 (Figure 1), we also identified compounds 53 and 54 (Figure 1) showing similar activities.…”

“…This observation agrees with our previous studies and with data reported in the literature, suggesting that the urea group is a crucial structural requirement to potently inhibit sEH. 20 Moreover, it is here demonstrated that the amide group was not able to give the network of H-bonds observed for the urea group. Indeed, we noted three hydrogen bonds in 3 (Figure S66B).…”

Antitumor Mechanisms of Lycium barbarum Fruit: An Overview of In Vitro and In Vivo Potential

“…The antioxidant activity of polyphenols is well known and results from a combination of their Fe ions chelating and oxidation-inhibiting properties, together with their inhibitory activity on oxidase enzymes widely involved in inflammatory diseases: 5-LOX, cyclooxygenase, soluble epoxide hydrolase, myeloperoxidase, NADPH oxidase, iNOS and xanthine oxidase [ 88 , 89 , 90 , 91 , 92 ].…”