Effects of semaglutide-loaded lipid nanocapsules on metabolic dysfunction-associated steatotic liver disease

Abstract

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a highly prevalent chronic liver disease that can progress to end-stage conditions with life-threatening complications, but no pharmacologic therapy has been approved. Drug delivery systems such as lipid nanocapsules (LNC) are very versatile platforms that are easy to produce and can induce the secretion of the native glucagon-like peptide 1 (GLP-1) when orally administered. GLP-1 analogs are currently being studied in clinical trials in the context of MASLD. Our nanosystem provides with increased levels of the native GLP-1 and increased plasmatic absorption of the encapsulated GLP-1 analog (semaglutide). Our goal was to use our strategy to demonstrate a better outcome and a greater impact on the metabolic syndrome associated with MASLD and on liver disease progression with our strategy compared with the oral marketed version of semaglutide, Rybelsus^®. Therefore, we studied the effect of our nanocarriers on a dietary mouse model of MASLD, the Western diet model, during a daily chronic treatment of 4 weeks. Overall, the results showed a positive impact of semaglutide-loaded lipid nanocapsules towards the normalization of glucose homeostasis and insulin resistance. In the liver, there were no significant changes in lipid accumulation, but an improvement in markers related to inflammation was observed. Overall, our strategy had a positive trend on the metabolic syndrome and at reducing inflammation, mitigating the progression of the disease. Oral administration of the nanosystem was more efficient at preventing the progression of the disease to more severe states when compared to the administration of Rybelsus^®, as a suspension.

Keywords: GLP-1 analogs; MASH; MASLD; Rybelsus; Semaglutide.

Fig. 1

SEMA-RM-LNC has a greater impact on the metabolic syndrome than RM-LNC and Rybelsus^® under non-fasting conditions throughout the one-month treatment. A Schematic representation of the treatment period of 4 weeks, B Body weight (%), C Pre/Post: Body weight (%), D Body weight change (%), E Non-fasting glucose (%), F Pre/Post: Non-fasting glucose (%), G Non-fasting glucose change (%), H Active GLP-1 levels (pg/mL) measured in portal plasma, I Total GLP-1 levels (pg/mL) measured in cava plasma. Pre: beginning of treatment; Post: end of treatment. The results in D, G were calculated by subtracting the post values from the pre values. P values in H, I were determined by One-way Anova followed by Tukey’s post hoc test or the Kruskal-Wallis followed by Dunn’s post hoc test (*P < 0.05, **P < 0.01, ****P < 0.0001). The data are presented as the mean ± SEM (n = 9–10)

Fig. 2

SEMA-RM-LNC has a greater impact on the metabolic syndrome than EXE-RM-LNC, RM-LNC and Rybelsus^® under fasting conditions throughout the one-month treatment A Schematic representation of the conduction of experiments under fasting conditions, B Fasting glucose (mg/dL), C Fasting insulin levels (ng/mL), D Homeostatic Model Assessment of Insulin Resistance (HOMA-IR) calculated using the equation [fasting glucose (mg/dL) x fasting insulin (ng/mL)/405], E Pre/Post: Fasting glucose (mg/dL), Pre/Post: Fasting insulin levels (ng/mL), G Pre/Post: HOMA-IR, H Fasting glucose change. The results in H were calculated by subtracting the post values from the pre values. P values in B were determined by One-way Anova followed by Tukey’s post hoc test. P values in E were determined by Two-way Anova followed by Tukey’s post hoc test (*P < 0.05, ***P < 0.001, ****P < 0.0001). The data are presented as the mean ± SEM (n = 9–10)

Fig. 3

SEMA-RM-LNC has a similar effect on lipid homeostasis than EXE-RM-LNC, RM-LNC and Rybelsus^® A Liver weight (g), B ALT levels (U/L) measured in systemic plasma, C AST levels (U/L) measured in systemic plasma, D Total lipid content per whole liver, E Histological NAFLD activity score (NAS), F Steatosis, ballooning and lobular inflammation individual scores, G Inflammatory foci per 20× field, H Representative H&E liver sections (scale bar: 100 μm). P values were determined by One-way Anova followed by Tukey’s post hoc test or Kruskal-Wallis followed by Dunn’s post hoc test. P values in F were determined by Two-way Anova followed by Tukey’s post hoc test (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001). The data are presented as the mean ± SEM (n = 9–10)

Fig. 4

SEMA-RM-LNC reduces inflammation and infiltration/recruitment of immune cell populations in the liver A Representative LY-6G staining of liver sections per group (scale bar: 50 μm) B Quantification of neutrophils in liver sections, C Heatmap representation of the relative mRNA expression normalized to the CTRL ND group. P values were determined by One-way Anova followed by Tukey’s post hoc test or by Kruskal-Wallis followed by Dunn’s post hoc test (*P < 0.05, ****P < 0.0001). The data are presented as the mean ± SEM (n = 9–10)