SECUAH 2021

dianas | Vol 8 No 1 | marzo 2019 | e201903p07

Novel anti-inflammatory agents via inhibition of NF-kappaB and/or Inflammasome pathways: a series of hispanolone-derived diterpenes

L González-cofrade, I Cuadrado, A Estévez-Braun, S Hortelano, B. Las Heras

Departamento de Farmacología, Farmacognosia y Botánica. Facultad de Farmacia, Universidad Complutense de Madrid

lagonz11@ucm.es; lauragoncof@gmail.com

IV Congreso de Señalización Celular, SECUAH 2019.
20-22 de marzo, 2019. Universidad de Alcalá. Alcalá de Henares, Madrid. España
Sesión de paneles.

Keywords: Inflammation; diterpenes; NF-κB; MAPKs; inflammasome

Abstract

Inflammation is a crucial host response triggered by invading pathogens and injured tissue. Natural products have played a significant role in human health as a source of new drugs to prevent and treat inflammatory conditions. Diterpenes are bioactive natural products with great therapeutic potential and are considered very promising starting points for the development of new therapeutic agents. In this study, a series of seventeen novel hispanolone derivatives (N1-N17) were synthesized and evaluated for potential anti-inflammatory activity in J774A.1 macrophages. We focused on their potential as inhibitors of classical inflammatory pathways (NF-κB/MAPKs) and/or NLRP3 inflammasome activation. All compounds except N2 and N5 were non-cytotoxic, as revealed the MTT assay. To evaluate the effects of diterpenes on NF-κB pathway, macrophages were activated with lipopolysaccharide (LPS) in the absence or presence of these compounds. Inhibitory effects of the non-toxic derivatives on nitric oxide (NO) production were evaluated. Hispanolone derivatives N1 and N12 were the most active compounds (IC50 in the range 10-20 µM) so they were selected for further evaluation. NOS-2 and COX-2 expression were significantly inhibited, as observed by western blot. Moreover, the phosphorylation of mitogen-activated protein kinases (MAPKs) ERK1/2, p38 and JNK was also reduced by pre-incubation with N1 and N12. We also investigated their potential as regulators of NLRP3 inflammasome. Cell treatment with diterpenes N12, N13, N16 and N17 at 20 µM significantly reduced IL-1β secretion in LPS/ATP or LPS/Nigericin-stimulated macrophages. Western blot analyses also confirmed that these compounds inhibited IL-1β and cleaved caspase-1 protein expression. In conclusion, these results show the promising anti-inflammatory effects of some hispanolone derivatives, in particular N12, acting on a dual level. This diterpene not only inhibits NF-κB/MAPKs signalling pathways, but also regulates inflammasome activation.