Novel Activities of Select NSAID R-Enantiomers against Rac1 and Cdc42 GTPases

Abstract

Rho family GTPases (including Rac, Rho and Cdc42) collectively control cell proliferation, adhesion and migration and are of interest as functional therapeutic targets in numerous epithelial cancers. Based on high throughput screening of the Prestwick Chemical Library® and cheminformatics we identified the R-enantiomers of two approved drugs (naproxen and ketorolac) as inhibitors of Rac1 and Cdc42. The corresponding S-enantiomers are considered the active component in racemic drug formulations, acting as non-steroidal anti-inflammatory drugs (NSAIDs) with selective activity against cyclooxygenases. Here, we show that the S-enantiomers of naproxen and ketorolac are inactive against the GTPases. Additionally, more than twenty other NSAIDs lacked inhibitory action against the GTPases, establishing the selectivity of the two identified NSAIDs. R-naproxen was first identified as a lead compound and tested in parallel with its S-enantiomer and the non-chiral 6-methoxy-naphthalene acetic acid (active metabolite of nabumetone, another NSAID) as a structural series. Cheminformatics-based substructure analyses—using the rotationally constrained carboxylate in R-naproxen—led to identification of racemic [R/S] ketorolac as a suitable FDA-approved candidate. Cell based measurement of GTPase activity (in animal and human cell lines) demonstrated that the R-enantiomers specifically inhibit epidermal growth factor stimulated Rac1 and Cdc42 activation. The GTPase inhibitory effects of the R-enantiomers in cells largely mimic those of established Rac1 (NSC23766) and Cdc42 (CID2950007/ML141) specific inhibitors. Docking predicts that rotational constraints position the carboxylate moieties of the R-enantiomers to preferentially coordinate the magnesium ion, thereby destabilizing nucleotide binding to Rac1 and Cdc42. The S-enantiomers can be docked but are less favorably positioned in proximity to the magnesium. R-naproxen and R-ketorolac have potential for rapid translation and efficacy in the treatment of several epithelial cancer types on account of established human toxicity profiles and novel activities against Rho-family GTPases.

Retracted: MicroRNA‑224 promotes the sensitivity of osteosarcoma cells to cisplatin by targeting Rac1

“…Rac1 is a member of the Ras superfamily of Rho proteins, and it plays an essential role in many cellular processes, including mitogenesis, kinase cascade activation, transcriptional activation, DNA synthesis and cytoskeleton reorganization [37][38][39][40][41][42]. Rac1 overexpression has been found in various types of cancer, such as lung cancer, gastric cancer, pancreatic cancer, bladder cancer and breast cancer [43][44][45][46][47].…”

Characterization of a Dual Rac/Cdc42 Inhibitor MBQ-167 in Metastatic Cancer

“…Although other small molecule inhibitors, such as the NSC23766 derivative Aza-1 (inhibits both Rac and Cdc42) and CID2950007/ML141 (selective for Cdc42) are currently available, they are effective in the micromolar range (22–24). Our goal to develop a Rac/Cdc42 inhibitor with improved activities led to the identification of MBQ-167.…”

Rho GTPases in A549 and Caco-2 cells dominating the endocytic pathways of nanocarbons with different morphologies

“…[6][7][8] It has been reported that Rac1, RhoA, and Cdc42 are associated with actin reorganization, cell motility, cell-cell and cell-extracellular matrix adhesion as well as cell cycle progression, gene expression, and apoptosis. 9 Rac1 is activated in protrusion and ruffling; RhoA's activity is related to protrusion, tail retraction, and ruffling; Cdc42 not only takes part in the process of protrusion, filopodia, but also controls invadopodia disassembly at the Golgi. 10 However, the existing studies did not focus on the impact of these Rho GTPase members on cellular uptake of nanoparticles.…”