Genfit (France)

Abstract Background and Aims Acute-on-chronic liver failure (ACLF), a syndrome observed among patients with decompensated liver cirrhosis is characterised by an intense systemic inflammatory response, multiple organ system failure and high 28-day mortality. The loss of kidney function is the most common extra-hepatic organ failure; acute kidney injury (AKI) being reported in up to 92% of patients with ACLF (Napoleone et al 2022). The intense systemic inflammation occurring in ACLF, triggered by bacteria or endotoxins such as lipopolysaccharide (LPS) due to gut leakage, has been shown to be involved in the pathogenesis of organ dysfunction and/or failure. The FDA approved antiparasitic drug nitazoxanide (NTZ) presents hepatoprotective and anti-inflammatory properties, making it a promising therapeutic approach for the treatment of ACLF. Our aim was to assess the efficacy of NTZ to alleviate kidney injury and inflammation and restore renal function in preclinical models of ACLF. Method ACLF was triggered through an acute injection of LPS in rats with advanced liver injury induced by either bile duct ligation (BDL) or carbon tetrachloride (CCl4). NTZ (100 mg/kg) or vehicle were orally administered concomitantly or one hour before LPS injection. Serum and tissues were collected 3 or 24 hours post-LPS in BDL and CCl4 rats, respectively. To evaluate renal function, serum levels of cystatin C, creatinine and urea were measured. Renal transcriptome was assessed using RNA sequencing and pairwise comparisons were performed to identify differentially expressed genes. Results In the CCl4 + LPS-induced ACLF model, NTZ alleviated hepatocellular injury by reducing LPS-induced rise of AST (−100%, p = 0.03) and GGT (−99%, p = 0.014) at 24 h post-LPS. Compared to fibrotic rats (CCl4 alone), CCl4+LPS-induced ACLF rats presented alterations of kidney function as shown by elevated levels of serum creatinine (113 µM vs 50 µM) and serum urea (127 mg/dL vs 47 mg/dL), which were reduced upon NTZ administration (−99% creatinine, p = 0.03 and −96% urea, p = 0.0005). LPS injection in rats with BDL-induced liver fibrosis led to systemic inflammation as shown by increased serum pro-inflammatory cytokines levels at 3 h post-LPS, which were significantly reduced upon NTZ treatment: −99% for IL6 (p = 0.001) and −94% for TNF alpha (p = 0.006). Compared to BDL alone rats, BDL+LPS-induced ACLF rats also presented renal dysfunction as shown by elevated levels of cystatin-C (4.6 µg/mL vs 2 µg/mL, Fig. A). This renal impairment was associated with a significant remodeling of the renal transcriptome (Fig. B), with differentially expressed genes mainly related to innate immune response and pro-inflammatory signaling pathways. Treatment of rats with NTZ restored the LPS-induced level of cystatin C by 85% (p = 0.004) and the inflammatory gene signature in the kidney (Figure), with expression level of genes such as lipocalin 2 (LCN2), a marker of renal injury, being reduced. Interestingly, serum and renal levels of Ripk3, a marker of necroptotic cell death, was also alleviated by NTZ treatment. Conclusion A single oral administration of NTZ rapidly alleviates kidney inflammation and injury and prevents alteration of renal function in pre-clinical models of ACLF. By protecting the major extrahepatic organ failing in ACLF, in addition to improvement of other organ functions, these findings further support the investigation of NTZ therapy as a promising approach for ACLF treatment.

IntroductionElafibranor, a dual peroxisome proliferator-activated receptor (PPAR) alpha/delta agonist, is under investigation as a treatment for primary biliary cholangitis (PBC). In vitro studies demonstrated that elafibranor and its major active metabolite GFT1007 inhibit organic anion transporter polypeptide (OATP) 1B3. A phase I trial was conducted to evaluate the drug-drug interaction (DDI) potential of elafibranor as a perpetrator with atorvastatin, an OATP1B3, OATP1B1 and cytochrome P4503A (CYP3A) clinical substrate.MethodsIn this phase I, randomised, open-label trial (EudraCT: 2015–000723-88), healthy participants received single doses of atorvastatin 40 mg (Days 1 and 17) and once-daily doses of elafibranor 180 mg (Days 4–19) under fasting conditions. Pharmacokinetic (PK) parameters derived for each treatment included observed maximum plasma concentration (Cmax), area under the plasma concentration curve from administration up to the last quantifiable concentration at 72 hours (AUC0–72 h), and time to reach Cmax (tmax). All adverse events (AEs) were recorded.ResultsOf 25 white male participants, 24 completed the trial; median (range) age was 32.0 (19–45) years. There was no increase in the overall exposure of atorvastatin. Compared with atorvastatin administration alone, Cmax was 28% lower following co-administration with elafibranor, with a geometric mean ratio (GMR) of 72 (90% confidence interval [CI]: 60.1–87.2); mean (standard deviation [SD]): 19.7 (11.0) ng/mL vs 14.8 (11.4) ng/mL. Median (range) tmax increased from 0.67 (0.50–3.00) hours (h) to 1.00 (0.67–4.50) h. There was a small decrease of mean (SD) AUC0–72 h (12%) following co-administration, and the 90% CIs indicated similarity between the regimens: 81.3 (42.1) ng/mL*h vs 75.0 (51.5) ng/mL*h; GMR: 88 (90% CI: 82.2–93.3). Observed differences between the two regimens were not deemed clinically significant. A total of 16 non-serious and mild to moderate treatment-emergent AEs were reported by 11 participants; all were resolved without treatment discontinuation.ConclusionsCo-administration of atorvastatin and elafibranor 180 mg had minimal impact on the overall exposure of atorvastatin. Therefore, co-administration of elafibranor 80 mg (clinical dose) with OATP1B1, OATP1B3 and CYP3A substrates, such as other statins, is unlikely to cause clinically significant PK DDIs. Furthermore, co-administration of atorvastatin and elafibranor was well tolerated.

The aim of the study was to assess the effects of a high-fructose diet (HFrD) on skeletal muscle transcriptomic response in healthy offspring of patients with type 2 diabetes, a subgroup of individuals prone to metabolic disorders. Ten healthy normal weight first-degree relatives of type 2 diabetic patients were submitted to a HFrD (+3.5 g fructose/kg fat-free mass per day) during 7 days. A global transcriptomic analysis was performed on skeletal muscle biopsies combined with in vitro experiments using primary myotubes. Transcriptomic analysis highlighted profound effects on fatty acid oxidation and mitochondrial pathways supporting the whole-body metabolic shift with the preferential use of carbohydrates instead of lipids. Bioinformatics tools pointed out possible transcription factors orchestrating this genomic regulation, such as PPARα and NR4A2. In vitro experiments in human myotubes suggested an indirect action of fructose in skeletal muscle, which seemed to be independent from lactate, uric acid, or nitric oxide. This study shows therefore that a large cluster of genes related to energy metabolism, mitochondrial function, and lipid oxidation was downregulated after 7 days of HFrD, thus supporting the concept that overconsumption of fructose-containing foods could contribute to metabolic deterioration in humans.

Alzheimer’s disease (AD) is a neurodegenerative condition that leads to neuronal death and memory dysfunction. In the past, specific peroxisome proliferator-activated receptor (PPAR)γ-agonists, such as pioglitazone, have been tested with limited success to improve AD pathology. Here, we investigated the therapeutic efficacy of GFT1803, a novel potent PPAR agonist that activates all the three PPAR isoforms (α/δ/γ) in the APP/PS1 mouse model in comparison to the selective PPARγ-agonist pioglitazone. Both compounds showed similar brain/plasma partitioning ratios, although whole body and brain exposure to GFT1803 was significantly lower as compared to pioglitazone, at doses used in this study. Oral treatment of APP/PS1 mice with GFT1803 decreased microglial activation, amyloid β (Aβ) plaque area, Aβ levels in sodium dodecyl sulfate- and formic acid-soluble fractions in a concentration-dependent manner. With a single exception of Aβ38 and Aβ40 levels, measured by ELISA, these effects were not observed in mice treated with pioglitazone. Both ligands decreased glial fibrillary acidic protein (GFAP) expression to similar extent and did not affect ApoE expression. Finally, GFT1803 increased insulin-degrading enzyme expression. Analysis of spatial memory formation in the Morris water maze demonstrated that both compounds were able to partially revert the phenotype of APP/PS1 mice in comparison to wild-type mice with GFT1803 being most effective. As compared to pioglitazone, GFT1803 (pan-PPAR agonist) produced both quantitatively superior and qualitatively different therapeutic effects with respect to amyloid plaque burden, insoluble Aβ content, and neuroinflammation at significantly lower whole body and brain exposure rates.

M1-activated Macrophages (M1M) play a major role in atherosclerotic lesions of aortic arch, promoting proinflammatory response. In vivo trafficking of M1M in aortic plaques is therefore critical. M1M from bone marrow cell culture were magnetically labeled, using iron nanoparticles, intravenously injected and followed up with 3 day magnetic resonance imaging (MRI) in mice developing macrophage-laden atheroma (ApoE2 knock-in mice). M1M recruitment in aortic arch lesions was assessed both by MRI and histology. In all ApoE2 knock-in mice injected with labeled cells, high resolution MRI showed localized signal loss regions in the thickened aortic wall, with a maximal effect at day 2 (-34% +/- 7.3% P < 0.001 compared with baseline). This was confirmed with Prussian blue (iron) staining and corresponded to M1M (Major Histo-compatibility Complex II positive). Clear different intraplaque and adventitial dynamic distribution profiles of labeled cells were observed during the 3 days. M1M dynamic MRI is a promising marker to noninvasively assess the macrophage trafficking underlying aortic arch plaque progression.

The farnesoid X receptor (FXR, NR1H4) belongs to the nuclear receptor superfamily and is activated by bile acids such as chenodeoxycholic acid, or synthetic ligands such as GW4064. FXR is implicated in the regulation of bile acid, lipid, and carbohydrate metabolism. Posttranslational modifications regulating its activity have not been investigated yet. Here, we demonstrate that calcium-dependent protein kinase C (PKC) inhibition impairs ligand-mediated regulation of FXR target genes. Moreover, in a transactivation assay, we show that FXR transcriptional activity is modulated by PKC. Furthermore, phorbol 12-myristate 13-acetate , a PKC activator, induces the phosphorylation of endogenous FXR in HepG2 cells and PKCalpha phosphorylates in vitro FXR in its DNA-binding domain on S135 and S154. Mutation of S135 and S154 to alanine residues reduces in cell FXR phosphorylation. In contrast to wild-type FXR, mutant FXRS135AS154A displays an impaired PKCalpha-induced transactivation and a decreased ligand-dependent FXR transactivation. Finally, phosphorylation of FXR by PKC promotes the recruitment of peroxisomal proliferator-activated receptor gamma coactivator 1alpha. In conclusion, these findings show that the phosphorylation of FXR induced by PKCalpha directly modulates the ability of agonists to activate FXR.

Polyvalent carbohydrate-protein interactions occur frequently in biology, particularly in recognition events on cellular membranes. Collectively, they can be much stronger than corresponding monovalent interactions, rendering it difficult to control them with individual small molecules. Artificial macromolecules have been used as polyvalent ligands to inhibit polyvalent processes; however, both reproducible synthesis and appropriate characterization of such complex entities is demanding. Herein, we present an alternative concept avoiding conventional macromolecules. Small glycodendrimers which fulfill single molecule entity criteria self-assemble to form non-covalent nanoparticles. These particles-not the individual molecules-function as polyvalent ligands, efficiently inhibiting polyvalent processes both in vitro and in vivo. The synthesis and characterization of these glycodendrimers is described in detail. Furthermore, we report on the characterization of the non-covalent nanoparticles formed and on their biological evaluation.