Chrysin-based supramolecular cyclodextrin-calixarene drug delivery system: a novel approach for attenuating cardiac fibrosis in chronic diabetes

Abstract

Introduction: Cardiac fibrosis is strongly induced by diabetic conditions. Both chrysin (CHR) and calixarene OTX008, a specific inhibitor of galectin 1 (Gal-1), seem able to reduce transforming growth factor beta (TGF-β)/SMAD pro-fibrotic pathways, but their use is limited to their low solubility. Therefore, we formulated a dual-action supramolecular system, combining CHR with sulfobutylated β-cyclodextrin (SBECD) and OTX008 (SBECD + OTX + CHR). Here we aimed to test the anti-fibrotic effects of SBECD + OTX + CHR in hyperglycemic H9c2 cardiomyocytes and in a mouse model of chronic diabetes.Methods: H9c2 cardiomyocytes were exposed to normal (NG, 5.5 mM) or high glucose (HG, 33 mM) for 48 h, then treated with SBECD + OTX + CHR (containing OTX008 0.75–1.25–2.5 µM) or the single compounds for 6 days. TGF-β/SMAD pathways, Mitogen-Activated Protein Kinases (MAPKs) and Gal-1 levels were assayed by Enzyme-Linked Immunosorbent Assays (ELISAs) or Real-Time Quantitative Reverse Transcription Polymerase chain reaction (qRT-PCR). Adult CD1 male mice received a single intraperitoneal (i.p.) administration of streptozotocin (STZ) at a dosage of 102 mg/kg body weight. From the second week of diabetes, mice received 2 times/week the following i.p. treatments: OTX (5 mg/kg)-SBECD; OTX (5 mg/kg)-SBECD-CHR, SBECD-CHR, SBECD. After a 22-week period of diabetes, mice were euthanized and cardiac tissue used for tissue staining, ELISA, qRT-PCR aimed to analyse TGF-β/SMAD, extracellular matrix (ECM) components and Gal-1.Results: In H9c2 cells exposed to HG, SBECD + OTX + CHR significantly ameliorated the damaged morphology and reduced TGF-β1, its receptors (TGFβR1 and TGFβR2), SMAD2/4, MAPKs and Gal-1. Accordingly, these markers were reduced also in cardiac tissue from chronic diabetes, in which an amelioration of cardiac remodeling and ECM was evident. In both settings, SBECD + OTX + CHR was the most effective treatment compared to the other ones.Conclusion: The CHR-based supramolecular SBECD-calixarene drug delivery system, by enhancing the solubility and the bioavailability of both CHR and calixarene OTX008, and by combining their effects, showed a strong anti-fibrotic activity in rat cardiomyocytes and in cardiac tissue from mice with chronic diabetes. Also an improved cardiac tissue remodeling was evident. Therefore, new drug delivery system, which could be considered as a novel putative therapeutic strategy for the treatment of diabetes-induced cardiac fibrosis.

Chrysin directing an enhanced solubility through the formation of a supramolecular cyclodextrin-calixarene drug delivery system: a potential strategy in antifibrotic diabetes therapeutics

“…To this regard, its anti-fibrotic properties have been verified in H9c2 exposed to hyperglycemic conditions, while its safety and efficacy have been tested intraperitoneally in adult CD1 male mice with chronic diabetes. In this in vivo experimental setting, CHR-OTX008-SBECD lead to a significant reduction of cardiac fibrosis markers and cardiac tissue remodeling, without any toxicity, by improving both CHR and OTX008 solubility in water and bioavailability [31]. Further detailed in vitro and in vivo studies are essential to discern the specific cellular and molecular mechanisms driving its therapeutic potential.…”

Chrysin Directing an Enhanced Solubility through the Formation of a Supramolecular Cyclodextrin–Calixarene Drug Delivery System: A Potential Strategy in Antifibrotic Diabetes Therapeutics

“…In this regard, its antifibrotic properties have been verified in H9c2 exposed to hyperglycemic conditions, while its safety and efficacy have been tested intraperitoneally in adult CD1 male mice with chronic diabetes. In this in vivo experimental setting, CHR-OTX008-SBECD led to a significant reduction in cardiac fibrosis markers and cardiac tissue remodeling without any toxicity by improving both CHR and OTX008 solubility in water and bioavailability [ 31 ]. Further detailed in vitro and in vivo studies are essential to discern the specific cellular and molecular mechanisms driving its therapeutic potential.…”

Ginsenoside Rb1 reduces oxidative/carbonyl stress damage and dysfunction of RyR2 in the heart of streptozotocin-induced diabetic rats

“…Inhibition of oxidative and carbonyl stress and phosphorylation of RyR2 have been suggested to deeply evaluate the therapeutical targets in diabetic cardiomyopathy. Exploring the possibility of new use drug therapies has the potential to identify treatments for comorbid conditions that have the added benefit of glycemic control [ 19 – 21 ]. To the best of our knowledge, rare medicines reduce both oxidative stress and dysfunction in RyR2.…”