Bromo-substituted indirubins for inhibition of protein kinase-mediated signalling involved in inflammatory mediator release in human monocytes.

Targeting protein kinases that regulate signalling pathways in inflammation is an effective pharmacological approach to alleviate uncontrolled inflammatory diseases. In this context, the natural product indirubin and its 6-bromo-substituted analogue 6-bromoindirubin-3 -glycerol-oxime ether (6BIGOE; 1) were identified as potent inhibitors of glycogen synthase kinase-3ß (GSK-3ß). These inhibitors suppress the release of pro-inflammatory cytokines and prostaglandins (PG) from human monocytes. However, indirubin derivatives target several protein kinases such as cyclin-dependent kinases (CDKs) which has been a major concern for their application in inflammation therapy. Here, we report on a library of 13 5-bromo-substituted indirubin derivatives that have been designed to improve potency and target selectivity. Side-by-side comparison of reference compound 1 (6BIGOE) with 5-bromo derivatives revealed its isomer 2 (5BIGOE), as the most potent derivative able to supress pro-inflammatory cytokine and PG release in lipopolysaccharide-stimulated human monocytes. Analysis of protein kinase inhibition in intact monocytes, supported by our in silico findings, proposed higher selectivity of 1 for GSK-3ß inhibition with lesser potency against CDKs 8 and 9. In contrast, 2 supressed the activity of these CDKs with higher effectiveness than GSK-3ß, representing additional targets of indirubins within the inflammatory response. Encapsulation of 1 and 2 into polymer-based nanoparticles (NP) improved their pharmacological potential. In conclusion, the 5- and 6-brominated indirubins 1 and 2 as dual GSK-3ß and CDK8/9 inhibitors represent a novel concept for intervention with inflammatory disorders.

Importance of Demographic and Clinical Features in Evaluating the Severity of COVID-19 in Hospitalized Patients: A Serbian Retrospective Study in the First Pandemic Year.

The aim of this study is to determine the demographic and initial clinical characteristics of patients with COVID-19 and their importance in evaluating the severity of the disease. A retrospective study included patients suffering from COVID-19 who were hospitalized at The Department of Infectious Disease of the Clinical Hospital Center Pristina-Gracanica from the beginning of the pandemic until the end of 2020. We compared the symptoms of the disease, radiographic findings of pneumonia, laboratory parameters, duration of symptoms before admission, the difference in the need for certain therapies, and the presence of comorbidities between non-severe and severe groups of patients. Patients with a severe disease were statistically significantly older. Hypertension was significantly associated with severe clinical conditions. Radiographic findings of bilateral pneumonia on admission were much more frequent among the severe group, and these patients' need for oxygen support was significantly higher. Lower neutrophil and higher lymphocyte counts were statistically significant in the non-severe group. Biochemical parameters at admission also showed statistical significance between the examined groups. Based on our research, we can conclude that a complete overview of the patient, including demographic and laboratory parameters as perhaps the most significant attributes, can help doctors in the timely clinical assessment of patients and, thus, in the timely application of adequate therapeutic protocols in the treatment of COVID-19.