An assessment of the effects of novel anti-inflammatory compounds in cell based studies

NF-κB and AP-1 are transcription factors with an evolutionary conserved role in the triggering and coordination of both innate and adaptive immune responses. Since they regulate a large number of inflammatory genes, they are considered as potential targets for anti-inflammatory drugs. In the current study, natural SU182 and synthetic alkaloid compounds, SU331, SU432 and minor groove binder compounds AIK18/85/1 and AIK18/70 obtained from the library associated with University of Strathclyde CRUK-Small Molecule Drug Discovery (SMDD) were investigated for possible anti-inflammatory effects at μM concentrations. In NCTC2544 human keratinocyte cells stably transfected with either NF-κB- or AP-1-linked luciferase reporter plasmids, tumour necrosis factor-α (TNF-α) and phorbol-12-myristate- 13 acetate (PMA) well characterized stimuli for canonical NF-κB pathway induced NF-κB and AP-1 transcriptional activity respectively. However, all tested compounds inhibited NF-κB transcriptional activity; in particular AIK18/85/1 prevented the TNF-α-induced translocation of NF-κB (p65) to the nucleus assessed by indirect immunoflouresnce. This effect of AIK18/85/1 was also reflected in the significant reduction of nuclear extract NF-κB-DNA binding activity as detected by Electrophoresis Mobility Shift Assay (EMSA), but without affecting the degradation of IκBα protein induced by TNF-α. Furthermore, AIK18/70 also decreased TNF-α- induced NF-κB-DNA binding activity but neither affected the phosphorylation of p65 nor the degradation of IκBα. On the other hand, both SU331 and SU432 inhibited the IκBα loss and resultant NF-κB-DNA binding activity in a concentration dependent manner. Although none of these compounds inhibited TNF-α-induced phosphorylation of NF-κB (Ser536-p65), their mode of inhibition on NF-κB signalling was sufficient to prevent the expression of NF-κB dependent proteins such as COX-2 and iNOS in LPS stimulated RAW 264.7 macrophage cells. Intriguingly, in contrast to other compounds SU182 was only effective at the level of NF-κB and AP-1 transcriptional activities, but without affecting the expression level of iNOS and COX-2 enzymes. Taken together these data indicate the potential for tested compounds to interfere with NF-κB signalling as IKK inhibitors or novel translocation inhibitor and thus may considered to be useful leads for the development of novel anti-inflammatory and anticancer drugs.

Name dc:type.qualificationname

phd

Level dc:type.qualificationlevel

doctoral-pg

Grantor dc:publisher.institution

University of Strathclyde

Year dc:date.issued

2012

Author dc:creator

• Gafri, Fadia Mohamed

DOI

10.48730/gwv9-kr25

Identifier

T13435

OAI identifier oai:identifier

oai:strathclyde:tq57nr067