Piperazine Derivatives Enhance Epithelial Cell Monolayer Permeability by Increased Cell Force Generation and Loss of Cadherin Structures

Abstract

A major obstacle for topical and enteral drug delivery is the poor transport of macromolecular drugs through the epithelium. One potential solution is the use of permeation enhancers that alter epithelial structures. Piperazine derivatives are known permeation enhancers that modulate epithelial structures, reduce transepithelial electrical resistance, and augment the absorption of macromolecular drugs. The mechanism by which piperazine derivatives disrupt the structures of epithelial monolayers is not well understood. Here, the effects of 1-phenylpiperazine and 1-methyl-4-phenylpiperazine are modeled in the epithelial cell line NRK-52E. Live-cell imaging reveals a dose-dependent gross reorganization of monolayers at high concentrations, but reorganization differs based on the piperazine molecule. Results show that low concentrations of piperazine derivatives increase myosin force generation within the cells and do not disrupt the cytoskeletal structure. Also, cytoskeletally attached cadherin junctions are disrupted before tight junctions. In summary, piperazines appear to increase myosin-mediated contraction followed by disruption of cell–cell contacts. These results provide new mechanistic insight into how transient epithelial permeation enhancers act and will inform of the development of future generations of transepithelial delivery systems.

Long‐term daily oral administration of intestinal permeation enhancers is safe and effective in mice

“…PPZ has been shown to affect paracellular permeability in previous work using in vitro and ex vivo models, and the gene expression data collected in this work allows us to extend our mechanistic understanding in vivo 7,20–22,44 . After one dose of enhancer, PPZ significantly decreased the expression of claudin 3 and ZO‐1 in the small intestine, and claudin 3 , ZO‐1 , and JAM‐A in the colon.…”

“…PPZ has been shown to affect paracellular permeability in previous work using in vitro and ex vivo models, and the gene expression data collected in this work allows us to extend our mechanistic understanding in vivo. 7,[20][21][22]44 After one dose of enhancer, PPZ significantly decreased the expression of claudin 3 and ZO-1 in the small intestine, and claudin 3, ZO-1, and JAM-A in the colon. These differences mirror observations in Caco-2 monolayers and ex vivo rat tissue that PPZ causes rearrangement of ZO-1 and actin, a cytoskeletal protein that extensively contacts the tight junctions via ZO-1.…”

Imaging methods in mechanosensing: a historical perspective and visions for the future

“…A previously unexplored route for mechanosensitive imaging technologies would be to examine the temporal impacts on cellular structure of small molecule drugs. In our recent work, we demonstrated that derivatives of piperazine, one of the earliest therapeutics for treating helminth infections ( Page, 2008 ), actively induce contractility in the actin cytoskeleton ( Zheng et al. , 2020 ).…”

“…, 2020 ). Along with other cellular effects, actomyosin contractility resulted in disassembly of cadherin junctions and the appearance of pores in the monolayer ( Zheng et al. , 2020 ).…”

Synthesis, Kinetics and Computational Explorations of 4‐Phenylpiperazine Bearing N‐(Aryl)‐3‐substituted‐benzamides as Auspicious Tyrosinase Inhibitors

“…[15][16] Recently, having distinction in pharmacological applications, there has been a need to synthesize compounds having 1,4substituted-piperazine and particularly N-phenylpiperazine, because such molecules displayed destitute cytotoxicity and have been useful to develop permeation, since they increase the absorptivity of the macromolecules across intestinal barricades and skin. [17][18] Benzamide is a derivative of benzoic acid and is considered the central hook of medicinal compounds. Several pharmacological activities of benzamides such as anticancer, cardiovascular, anti-HBV, neurotoxicity, anticonvulsant and CNS depressant have been reported.…”