Diclofenac N-Derivatives as Therapeutic Agents with Anti-Inflammatory and Anti-Cancer Effect
Creators: name: Galisteo, Alberto
name: Jannus, Fatin
name: García-García, Amalia
name: Aheget, Houssam
name: Rojas, Sara
name: Lupiañez, José A.
name: Rodríguez-Diéguez, Antonio
name: Reyes-Zurita, Fernando J.
name: Moral, José F. Quílez del
Publisher: name: figshare
Year: 2021
Subjects: subject: Biochemistry
subject: Organic Chemistry
subject: FOS: Chemical sciences
subject: Cancer
subject: Cell Biology

description: A series of diclofenac N-derivatives (2, 4, 6, 8c, 9c, 10a-c) were synthesized in order to testtheir anti-cancer and anti-inflammatory effects. The anticarcinogen activity has been assayed againstthree cancer cell lines: HT29, human colon cancer cells; Hep-G2, human hepatic cells; and B16-F10,murine melanoma cells. First, we determined the cytotoxicity of the different compounds, findingthat the most effective compound was compound 8c against all cell lines and both compounds 4 and6 in human Hep-G2 and HT29 cell lines. Compounds 4 and 8c were selected for the percentage ofapoptosis determination, cell cycle distribution, and mitochondrial membrane potential measurebecause these products presented the lowest IC50 values in two of the three cancer cell lines assayed(B16-F10 and HepG2), and were two of the three products with lowest IC50 in HT29 cell line.Moreover, the percentages of apoptosis induction were determined for compounds 4 and 8c, showingthat the highest values were between 30 to 60%. Next, the effects of these two compounds wereobserved on the cellular cycle, resulting in an increase in the cell population in G2/M cell cycle phaseafter treatment with product 8c, whereas compound 4 increased the cells in phase G0/G1, by possibledifferentiation process induction. Finally, to determine the possible apoptosis mechanism triggeredby these compounds, mitochondrial potential was evaluated, indicating the possible activation ofextrinsic apoptotic mechanism. On the other hand, we studied the anti-inflammatory effects ofthese diclofenac (DCF) derivatives on lipopolysaccharide (LPS) activated RAW264.7 macrophagesmonocytesmurine cells by inhibition of nitric oxide (NO) production. As a first step, we determinedthe cytotoxicity of the synthesized compounds, as well as DCF, against these cells. Then, sub-cytotoxicconcentrations were used to determine NO release at different incubation times. The greatest antiinflammatoryeffect was observed for products 2, 4, 8c, 10a, 10b, and 9c at 20 ?g?mL􀀀1 concentrationafter 48 h of treatment, with inhibition of produced NO between 60 to 75%, and a concentration thatreduces to the 50% the production of NO (IC50 NO) between 2.5 to 25 times lower than that of DCF. Inthis work, we synthesized and determined for the first time the anti-cancer and anti-inflammatorypotential of eight diclofenac N-derivatives. In agreement with the recent evidences suggesting thatinflammation may contribute to all states of tumorigenesis, the development of these new derivativescapable of inducing apoptosis and anti-inflammatory effects at very low concentrations representnew effective therapeutic strategies against these diseases.