A dual farnesoid X receptor/soluble epoxide hydrolase modulator treats non-alcoholic steatohepatitis in mice

Abstract

Non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH) are the most prevalent metabolic liver disorders and a serious global health burden. NAFLD/NASH pathogenesis and progression are highly multi-factorial and likely demand a combination of multiple mechanisms to provide a more effective treatment. We have developed a dual farnesoid X receptor agonist (FXRA)/soluble epoxide hydrolase inhibitor (sEHi) to simultaneously address two validated and complementary modes of action in NASH treatment. Here we report the in vivo profiling for this FXRA/sEHi in toxin-and diet-induced rodent NASH models. In streptozotocininduced NASH as a proof-of-concept study, the experimental FXRA/sEHi drug robustly prevented hepatic steatosis and fibrosis, and improved lipid homeostasis as well as biochemical markers of liver health. In methionine-choline-deficient high-fat diet-induced NASH, FXRA/sEHi treatment reduced hepatic steatosis and fibrosis to levels similar to healthy animals and demonstrated antiinflammatory activity confirming that dual FXRA/sEHi modulation produces a triad of complementary anti-NASH effects. Our results validate dual FXRA/sEHi modulation as an effective therapeutic strategy to treat NASH and advocates for a combinational drug therapeutic approach for multifactorial liver diseases.

Development and in vitro Profiling of Dual FXR/LTA4H Modulators

“…We then probed the effect of chain elongation from 3‐aminobenzoic acid ( 8 ) to 3‐aminophenylacetic acid ( 17 ) which was not favored by LTA4H, however, and caused a marked decrease in FXR activation activity. A methylsulfonamide motif ( 18 ) as carboxylic acid bioisostere, which had enabled the design of a highly potent dual FXR/sEH modulator,[ 6 , 11 ] did not alter potency on LTA4H but diminished FXR agonism, as well. In the phenoxy series, a 2‐aminoisonicotinic acid ( 15 ) head group was favored by both targets especially in terms of FXR activation efficacy.…”

“…Both compounds also did not inhibit soluble epoxide hydrolase (sEH), an enzyme of the arachidonic acid cascade which converts epoxyeicosatrienoic acids to the corresponding diols, at relevant concentrations (Table 4 ). This favorable selectivity profile is an important feature for FXRa/LTA4Hi tools to be used in experimental NASH treatment since inhibition of sEH has shown therapeutic efficacy in several NASH models,[ 11 , 32 , 33 ] too, affecting the outcome of pharmacological studies conducted with FXRa/LTA4Hi. Moreover, 19 caused no cytotoxic effects in a WST‐1 assay in HepG2 cells up to 100 μM concentration while 8 exhibited a slight anti‐proliferative activity at high concentration (Figure 2 b).…”

“…Following this strategy, we have previously developed dual modulators to target, for example, farnesoid X receptor (FXR) and soluble epoxide hydrolase (sEH), [6–8] or FXR and peroxisome proliferator‐activated receptors (PPARs) [9,10] . Strong therapeutic efficacy of such multi‐target agents in preclinical NASH models [10,11] corroborates the potential of designed polypharmacology in this indication. We further hypothesize that a combination of partial FXR activation [12] and inhibition of leukotriene A4 hydrolase (LTA4H) can efficiently counteract NAFLD/NASH.…”

Multi-Target Approaches in Metabolic Syndrome

“…The dual sEH/FXR modulator 7 reduced the hepatic steatosis and fibrosis. Furthermore, it exhibited a pronounced anti-inflammatory effect, thereby demonstrating excellent efficacy in the complex setting of NASH (Hye Khan et al, 2019).…”

Multi-Target Molecule to Treat Diabetic Nephropathy in Rats

“…Tissue sections were deparaffinized, re-hydrated, and kidney tissue slices were stained with Periodic Acid-Schiff (PAS) and Picrosirius Red (PSR). Glomerulosclerosis and mesangial matrix expansion were scored from kidney sections stained with PAS staining using methods described earlier (Hye Khan et al, 2018Khan et al, ,2019. Histological analysis was done at a magnification of 400X to assess glomerular injury, and renal tubular proteinaceous cast was assessed at a magnification of 200X using Nikon NIS Elements Software (Nikon Instruments Inc., Melville, NY, USA).…”