Discovery of Novel Orally Active Tetrahydro-Naphthyl-N-Acylhydrazones with In Vivo Anti-TNF-α Effect and Remarkable Anti-Inflammatory Properties

Abstract

LASSBio-1524 was designed as inhibitor of the IKK-β (kappa β kinase inhibitor) enzyme, which participates in the activation of the nuclear factor κB (NF-κB) canonical pathway, and its three N-acylhydrazone new analogues, LASSBio-1760, LASSBio-1763 and LASSBio-1764 are now being tested on their anti-inflammatory potential. The activity of these compounds was evaluated with the subcutaneous air pouch induced by carrageenan and by subsequent measurement of tumor necrosis factor-α (TNF-α), nitric oxide (NO) and reactive oxygen species (ROS). In the acute inflammation model, the oral pretreatment with doses from 0.3 to 30 mg/kg of N-acylhydrazone derivatives was able to significantly reduce leukocyte migration to the cavity. Pretreatment with LASSBio-1524 and its analogues also decreased NO, TNF-α and ROS biosynthesis an events closely involved with NF-kB pathway. The tetrahydronaphthyl-N-acylhydrazone derivative LASSBio-1764 was the most promising compound from this series, surpassing even LASSBio-1524. Additionally, none of the compounds demonstrated myelotoxicity or cytotoxicity. Cell viability was assayed and these compounds demonstrated to be safe at different concentrations. Western blot analysis demonstrated that LASSBio-1524 and LASSBio-1760 inhibited NF-κB expression in RAW 264.7 cell lineage. Our data indicate that the tested compounds have anti-inflammatory activity, which may be related to inhibition of leukocyte migration, reducing the production of NO, TNF-α and ROS. LASSBio-1524 and LASSBio-1760, in addition to these features, also reduced p65 nuclear expression assessed by western blot in RAW 264.7 murine cells.

Discovery of naphthyl‐N‐acylhydrazone p38α MAPK inhibitors with in vivo anti‐inflammatory and anti‐TNF‐α activity

“…All N ‐acylhydrazone derivatives ( 3a–d ) were obtained as a single diastereomer. 1 H‐NMR spectra of the compounds did not show any duplication in the amide and imine hydrogens of N ‐acylhydrazone subunit (Table ), that is a strong indication of the presence of the thermodynamically more stable ( E )‐diastereomer, as previously reported by our research group …”

Characterization of βN-Octadecanoyl-5-hydroxytryptamide Anti-Inflammatory Effect

“…Leukocytes collected from SAP were placed in tubes (1 × 10 6 cells/mL) and incubated for 1 h (37 °C and 5% CO 2 ). After incubation with C 18 -5HT (0.1, 0.3, and 1 µM) for 30 min and stimulated with phorbol myristate acetate (PMA, 10 nM) for 45 min, DCF-DA (2′-7′diclorodihidrofluorescein diacetate, 2 mM) was added, and the cells were incubated for 30 min (37 °C and 5% CO 2 ) [ 51 ]. The emitted fluorescence was captured in the FL-1 channel flow cytometer (BD Accuri™, B&D, Franklin Lakes, NJ, USA) and was expressed as DCF-DA fluorescence.…”

Characterization of anti-inflammatory effect and possible mechanism of action of Tibouchina granulosa

“…The amount of reactive oxygen species (ROS) produced by SAP leucocytes was measured as described in Cordeiro et al . Briefly, leucocytes collected in SAP were placed in tubes (10 6 cells) in a volume of 1 ml.…”