# Evaluation of the gastrotolerability of ketoprofen, lysine, and gabapentin co-crystal administration in an in vitro model of gastric epithelium: a proteomic update

## TL;DR

This study evaluates how well a new drug combination of ketoprofen, lysine, and gabapentin is tolerated in the stomach using a lab model.

## Contribution

The study introduces a new ternary drug co-crystal and provides proteomic evidence of its improved gastric tolerability.

## Key findings

- The KLS-GABA co-crystal showed higher gastric tolerability compared to individual or combined drug administration.
- Proteomic analysis revealed modulation of oxidative stress-related proteins like GSTP1.
- 2DE and western blot confirmed differential proteomic changes in the gastric epithelium model.

## Abstract

Chronic pain is a distressful condition that impacts strongly on people’s health, and, to date, no cure has been found. However, several strategies against pain have been proposed. Promising data regarding the usage of nonsteroidal anti-inflammatory drugs (NSAIDs) in combination with gabapentin in pain management laid the foundations for more complex approaches. A recently published study proposed a multimodal approach based on ketoprofen lysine salt (KLS) combined with gabapentin (GABA) in the context of chronic pain. Experiments on in vitro models showed supra-additive effects in modulating key pathways involved in neuropathic pain and gastric mucosal damage. Thus, the co-crystallization of ketoprofen, lysine, and gabapentin led to a new ternary drug-drug co-crystal (KLS-GABA co-crystal) to better take advantage of such effects. The new compound showed positive features in in vitro and in vivo pain models, particularly at the gastrointestinal level. To better understand the gastric impact of the co-crystal we chose to analyze proteomic fluctuations that occur in an in vitro model of gastric epithelium upon ethanol injury, aiming to observe the gastric effects of KLS-GABA co-crystal’s administration in comparison with KLS or GABA alone or co-administered as in the multimodal approach. Thus, we performed a 2-dimensional gel electrophoresis (2DE) to compare proteomes from lysates of NCI-N87 cells, chosen as model of gastric epithelium. Among all the localized spots (n = 117), the differentially abundant ones have been filtered and excised (n = 24) to perform mass spectrometry. A total of 414 non-redundant proteins have been found in the excised spots analyzed. A Gene Ontology-based enrichment analysis identified the proteins involved in biological processes, cellular components, and pathways. We then compared the 2DE findings with the western blot analysis confirming the differential proteomic fluctuations in the model. The methodology described here provides a broader picture of the effects of KLS, GABA, and KLS-GABA co-crystal administration in the ethanol-gastric injury model, identifying processes not revealed by other studies by showing proteomic changes and related mechanisms in detail, particularly via modulation of the oxidative stress-related GSTP1 which suggests the higher gastric tolerability of KLS-GABA co-crystal in the analyzed model highlighting its clinical reliability.