INH14

N‐(4‐Ethylphenyl)‐N′‐phenylurea (INH14) is a fragment‐like compound that inhibits the toll‐like receptor 2 (TLR2)‐mediated inflammatory activity and other inflammatory pathways (i.e., TLR4, TNF‐R and IL‐1R). In this study, we determined the molecular target of INH14. Overexpression of proteins that are part of the TLR2 pathway in cells treated with INH14 indicated that the target lay downstream of the complex TAK1/TAB1. Immunoblot assays showed that INH14 decreased IkBα degradation in cells activated by lipopeptide (TLR2 ligand). These data indicated the kinases IKKα and/or IKKβ as the targets of INH14, which was confirmed with kinase assays (IC50 IKKα=8.97 μm; IC50 IKKβ=3.59 μm). Furthermore, in vivo experiments showed that INH14 decreased TNFα formed after lipopeptide‐induced inflammation, and treatment of ovarian cancer cells with INH14 led to a reduction of NF‐kB constitutive activity and a reduction in the wound‐closing ability of these cells. These results demonstrate that INH14 decreases NF‐kB activation through the inhibition of IKKs. Optimization of INH14 could lead to potent inhibitors of IKKs that might be used as antiinflammatory drugs.

Herein we report the synthesis and anticonvulsant activity of a library of eighteen new compounds that are structural mimics of phenytoin. These class of compounds contain a N-1', N-3'-disubstituted spirohydantoin scaffold, where the N-1' and N-3' positions are modified with an alkyl group or aryl group. Of the eighteen compounds synthesized and tested, compound 5c showed the best anticonvulsant activity. It completely prevented the precursor events of motor seizure in the pilocarpine model of temporal lobe epilepsy. Additionally, ten of the analogs were more effective than phenytoin when compared using the Racine's score in the pilocarpine model. Based on the structure activity relationship (SAR), we concluded that alkyl groups (ethyl, propyl or cyclopropyl) at N-3' position and 4-nitro phenyl group at N-1' position are desirable.