Highly Selective Drug-Derived Fluorescent Probes for the Cannabinoid Receptor Type 1 (CB1R).

The cannabinoid receptor type 1 (CB1R) is pivotal within the endocannabinoid system regulating various signaling cascades with effects in appetite regulation, pain perception, memory formation, and thermoregulation. Still, understanding of CB1R's cellular signaling, distribution, and expression dynamics is very fragmentary. Real-time visualization of CB1R is crucial for addressing these questions. Selective drug-like CB1R ligands with a defined pharmacological profile were investigated for the construction of CB1R fluorescent probes using a reverse design-approach. A modular design concept with a diethyl glycine-based building block as the centerpiece allowed for the straightforward synthesis of novel probe candidates. Validated by computational docking studies, radioligand binding, and cAMP assay, this systematic approach allowed for the identification of novel pyrrole-based CB1R fluorescent probes. Application in fluorescence-based target-engagement studies and live cell imaging exemplify the great versatility of the tailored CB1R probes for investigating CB1R localization, trafficking, pharmacology, and its pathological implications.

Structure-Based Design of Xanthine-Imidazopyridines and -Imidazothiazoles as Highly Potent and In Vivo Efficacious Tryptophan Hydroxylase Inhibitors.

Tryptophan hydroxylases catalyze the first and rate-limiting step in the biosynthesis of serotonin, a well-known neurotransmitter that plays an important role in multiple physiological functions. A reduction of serotonin levels, especially in the brain, can cause dysregulation leading to depression or insomnia. In contrast, overproduction of peripheral serotonin is associated with symptoms like carcinoid syndrome and pulmonary arterial hypertension. Recently, we developed a class of TPH inhibitors based on xanthine-benzimidazoles, characterized by a tripartite-binding mode spanning the binding sites of the cosubstrate pterin and the substrate tryptophan and by chelation of the catalytic iron ion. Herein, we describe the structure-based development of a second generation of xanthine-imidiazopyridines and -imidazothiazoles designed to inhibit TPH1 in the periphery while preventing the interaction with TPH2 in the brain. Lead compound 32 (TPT-004) shows superior pharmacokinetic and pharmacodynamic properties as well as efficacy in preclinical models of peripheral serotonin attenuation and colorectal tumor growth.

Influence of Training and Single Exercise on Leptin Level and Metabolism in Obese Overweight and Normal-Weight Women of Different Age.

Leptin is one of the important hormones secreted by adipose tissue. It participates in the regulation of energy processes in the body through central and peripheral mechanisms. The aim of this study was to analyse the anthropological and physical performance changes during 9 month training in women of different age and body mass. The additional aim was the analysis of leptin levels in the fasting stage and after a control exercise. Obese (O), overweight (OW), and normal-weight (N) women participated in the study. Additional subgroups of premenopausal (PRE) (<50 years) and postmenopausal (POST) (50+) women were created for leptin level analysis. The main criterion of the division into subgroups was the age of menopause in the population. The control submaximal test and maximal oxygen uptake (VO2max) according to Astrand-Rhyming procedures was performed at baseline and after 3, 6, and 9 months. Before each control test, body weight (BM), body mass index (BMI), percentage of adipose tissue (% FAT), and mass (FAT (kg)) were measured. Moreover, before and after each test, leptin level was measured. After 9 months, there was a significant decrease in BM in the O (p < 0.05) and OW (p < 0.05) groups with no significant changes in the N group. There was a decrease in BMI in both the O (p < 0.05) and the OW (p < 0.05) groups, with no changes in the N group. The % FAT reduction was noted only in the O group (p < 0.05). VO2max increased in each of the measured groups (p < 0.05). The fasting leptin level at 0, 3, 6, and 9 months were the highest in the O group. The fasting leptin level before training was highest in the O group compared to the OW group (p < 0.01) and the N group (p < 0.01). It was also higher in the OW group compared to the N group at baseline (0) (p < 0.01) and after 3 and 6 months (p < 0.01). After 9 months, the leptin concentration decreased by 20.2% in the O group, 40.7% in the OW group, and 33% in the N group. Moreover, the fasting leptin level was higher in the POST subgroup compared to the PRE group in the whole group of women (p < 0.05). After a single exercise, the level of leptin in the whole study group decreased (p < 0.05). This was clearly seen, especially in the POST group. The 9 month training had a reducing effect on the blood leptin concentration in groups O, OW, and N. This may have been a result of weight loss and the percentage of fat in the body, as well as systematically disturbed energy homeostasis.

Detection of cannabinoid receptor type 2 in native cells and zebrafish with a highly potent, cell-permeable fluorescent probe.

Despite its essential role in the (patho)physiology of several diseases, CB2R tissue expression profiles and signaling mechanisms are not yet fully understood. We report the development of a highly potent, fluorescent CB2R agonist probe employing structure-based reverse design. It commences with a highly potent, preclinically validated ligand, which is conjugated to a silicon-rhodamine fluorophore, enabling cell permeability. The probe is the first to preserve interspecies affinity and selectivity for both mouse and human CB2R. Extensive cross-validation (FACS, TR-FRET and confocal microscopy) set the stage for CB2R detection in endogenously expressing living cells along with zebrafish larvae. Together, these findings will benefit clinical translatability of CB2R based drugs.

Discovery and development of CPL207280 as new GPR40/FFA1 agonist.

Due to a unique mechanism that limits the possibility of hypoglycemia, the free fatty acid receptor (FFA1) is an attractive target for the treatment of type 2 diabetes. So far, however, none of the promising agonists have been able to enter the market. The most advanced clinical candidate, TAK-875, was withdrawn from phase III clinical trials due to liver safety issues. In this article, we describe the key aspects leading to the discovery of CPL207280 (13), the design of which focused on long-term safety. The introduction of small, nature-inspired acyclic structural fragments resulted in compounds with retained high potency and a satisfactory pharmacokinetic profile. Optimized synthesis and upscaling provided a stable, solid form of CPL207280-51 (45) with the properties required for the toxicology studies and ongoing clinical trials.

A Palladium-Catalyzed Ullmann Cross-Coupling/Reductive Cyclization Route to the Carbazole Natural Products 3-Methyl-9H-carbazole, Glycoborine, Glycozoline, Clauszoline K, Mukonine, and Karapinchamine A.

The title natural products 2-7 have been prepared by reductive cyclization of the relevant 2-arylcyclohex-2-en-1-one (e.g. 20) to the corresponding tetrahydrocarbazole and dehydrogenation (aromatization) of this to give the target carbazole (e.g. 4). Compounds such as 20 were prepared using a palladium-catalyzed Ullmann cross-coupling reaction between the appropriate 2-iodocyclohex-2-en-1-one and o-halonitrobenzene.

Physical activity versus alcohol consumption among teenagers in chosen middle schools from cities central and south-western Poland.

BACKGROUND: Alcohol consumption is becoming increasingly popular among teenagers in Poland and affects younger and younger people. Many government promotional campaigns are aimed at reducing this phenomenon. There is an opinion that encouraging young people to practice sports and promoting a healthy lifestyle through sports and physical recreation could be helpful in solving this problem. OBJECTIVES: The aim of this study was to determine the relationship between physical activity declared by the Junior High Schools students and alcohol consumption. MATERIAL AND METHODS: The study was carried out on a group of 450 students from six Junior High Schools from central and south-western Poland. The Core Questions Global Youth Tobacco Survey (GYTS) - extended by authors of this paper with the descriptive part on the declared level of physical activity - was used to survey the teenagers. RESULTS: Physical activity is an important factor influencing alcohol consumption and consumption frequency by Junior High Schools students. The study has shown that physical activity is related to alcohol consumption declared by the Junior High Schools students - the number of days they drink alcohol and the number of days when they are drunk, particularly girls. In fact, girls are more prone to consuming large amounts of alcohol and get drunk, especially when they belong to a group of physically passive subjects. The lowest alcohol consumption has been observed in Junior High Schools students, who had declared irregular physical activity, but the highest and the most frequent one has been observed in students, who had declared physical passivity. The students, mostly boys, who declared the regular physical activity (they most commonly practiced sports), had not abstained from alcohol, especially beer and had got drunk often. CONCLUSIONS: The physical activity may be an important factor in preventing excessive alcohol consumption. Participation in physical activity is beneficial especially to girls, since when they remain physically passive, they often turn to alcohol. However, one should take into account the type of physical activity and the nature of the effort, as the authors of similar studies point out.