Identification of the Natural Steroid Sapogenin Diosgenin as a Direct Dual-Specific RORα/γ Inverse Agonist.

The steroid sapogenin diosgenin is a well-known natural product with a plethora of described pharmacological activities including the amelioration of T helper 17 (Th17)-driven pathologies. However, the exact underlying mode of action of diosgenin leading to a dampened Th17 response is still largely unknown and specific molecular targets have yet to be identified. Here, we show that diosgenin acts as a direct ligand and inverse agonist of the nuclear receptor retinoic acid receptor (RAR)-related orphan receptor (ROR)α and RORγ, which are key transcription factors involved in Th17 cell differentiation and metabolism. IC50 values determined by luciferase reporter gene assays, employing constructs for either RORγ-Gal4 fusion proteins or full length receptors, were in the low micromolar range at around 2 µM. To highlight the functional consequences of this RORα/γ inverse agonism, we determined gene expression levels of important ROR target genes, i.e., IL-17A and glucose-6-phosphatase, in relevant cellular in vitro models of Jurkat T and HepG2 cells, respectively, by RT-qPCR (reverse transcription quantitative PCR). Thereby, it was shown that diosgenin leads to a dose-dependent decrease in target gene expressions consistent with its potent cellular ROR inverse agonistic activity. Additionally, in silico dockings of diosgenin to the ROR ligand-binding domain were performed to determine the underlying binding mode. Taken together, our results establish diosgenin as a novel, direct and dual-selective RORα/γ inverse agonist. This finding establishes a direct molecular target for diosgenin for the first time, which can further explain reported amendments in Th17-driven diseases by this compound.

Natural products as modulators of retinoic acid receptor-related orphan receptors (RORs).

Covering: 1994 to 2020 Retinoic acid receptor-related orphan receptors (RORs) belong to a subfamily of the nuclear receptor superfamily and possess prominent roles in circadian rhythm, metabolism, inflammation, and cancer. They have been subject of research for over two decades and represent attractive but challenging drug targets. Natural products were among the first identified ligands of RORs and continue to be of interest to this day. This review focuses on ligands and indirect modulators of RORs from natural sources and explores their roles in a therapeutic context.

Trimethoprim-Loaded PLGA Nanoparticles Grafted with WGA as Potential Intravesical Therapy of Urinary Tract Infections-Studies on Adhesion to SV-HUCs Under Varying Time, pH, and Drug-Loading Conditions.

Intravesical therapy, already used to treat bladder cancer, is a potential treatment option for urinary tract infections. However, short dwelling time and washout proved to be challenging obstacles. To circumvent these issues, PLGA 503H and PLGA 2300 nanoparticles were prepared and surface modified with wheat germ agglutinin (WGA). Nanoparticles of both poly(d,l-lactic-co-glycolic acid) (PLGA) types exhibited high inherent adhesion to human uroepithelial cells. Although surface-bound WGA could be easily increased, adhesion did not. Loading the nanoparticles with trimethoprim did not counteract cell adhesion. Varying the medium for instillation revealed highest adhesion in sodium bicarbonate buffer (pH 5). To evaluate dwelling time, nanoparticles were incubated with the cell monolayer for increasing time intervals. A contact time of 15 min seems to be too short for adhesion to the cells as less than 50% particles remained bound after washing. However, after 30 min 70% of the particles added were bound, and afterward, no further increase was observed. WGA only slightly increased the adhesion of the PLGA nanoparticles, but this approach might not be economically viable. However, PLGA nanoparticles displayed a high inherent adhesion to cells that might substantially foster intravesical therapy.

Micro vs. nano: PLGA particles loaded with trimethoprim for instillative treatment of urinary tract infections.

Recurring infections and increasing resistances continue to complicate treatment of urinary tract infections. To investigate alternative treatment options, trimethoprim loaded micro- (D[4;3] of 1-9 µm) and nanoparticles (Z-Avg of 200-400 nm) were prepared from two types of poly(d,l-lactic-co-glycolic acid) (PLGA) for instillative therapy. While PLGA 503H microparticles could not be loaded with more than 2.6% trimethoprim, PLGA 2300 entrapped 22%. When preparing nanoparticles, both types displayed an even higher drug load of up to 29% using PLGA 2300, while PLGA 503H drug load stagnated at 10%. After eight hours, drug release from microparticles amounted to 55% (503H) and 35% (2300) whereas total drug release occurred after 8 (503H) and 9 days (2300). In case of nanoparticles, trimethoprim was liberated much faster with 60% after 2 h and a complete release after 24 h from both polymers. PLGA 2300 seems to be the better choice for entrapment of trimethoprim in microparticles considering the drug load. Both polymers, however, seem to be viable options for nanoparticles. Due to the higher overall drug load, nanoparticles seem to be advantageous over microparticles for instillative therapy, especially when prepared with PLGA 2300.