Exploring N-acyl-4-azatetracyclo[5.3.2.02,6.08,10]dodec-11-enes as 11?-HSD1 Inhibitors.
ABSTRACT: We recently found that a cyclohexanecarboxamide derived from 4-azatetracyclo[5.3.2.02,6.08,10]dodec-11-ene displayed low nanomolar inhibition of 11?-HSD1. In continuation of our efforts to discover potent and selective 11?-HSD1 inhibitors, herein we explored several replacements for the cyclohexane ring. Some derivatives exhibited potent inhibitory activity against human 11?-HSD1, although with low selectivity over the isoenzyme 11?-HSD2, and poor microsomal stability.
Project description:Glucocorticoid metabolism at the tissue level is regulated by two isoenzymes 11?-hydroxysteroid dehydrogenase (11?-HSD), which mutually convert biologically active cortisol and inactive cortisone. Recent research is focused on the role of 11?-HSD1 and 11?-HSD2 as autocrine factors of tumor cell proliferation and differentiation. Herein, we report the synthesis of novel 2-(isopropylamino)thiazol-4(5H)-one derivatives and their inhibitory activity for 11?-HSD1 and 11?-HSD2. The derivative containing the spiro system of thiazole and cyclohexane rings shows the highest degree of 11?-HSD1 inhibition (54.53% at 10 µM) and is the most selective inhibitor of this enzyme among the tested compounds. In turn, derivatives containing ethyl and n-propyl group at C-5 of thiazole ring inhibit the activity of 11?-HSD2 to a high degree (47.08 and 54.59% at 10 µM respectively) and are completely selective. Inhibition of the activity of these enzymes may have a significant impact on the process of formation and course of tumors. Therefore, these compounds can be considered as potential pharmaceuticals supporting anti-cancer therapy.
Project description:11?-hydroxysteroid dehydrogenase type?1 (11?-HSD1) plays a key role in converting intracellular cortisone to physiologically active cortisol, which is implicated in the development of several phenotypes of metabolic syndrome. Inhibition of 11?-HSD1 activity with selective inhibitors has beneficial effects on various conditions, including diabetes, dyslipidemia and obesity, and therefore constitutes a promising strategy to discover novel therapies for metabolic and cardiovascular diseases. A series of novel adamantyl heterocyclic ketones provides potent and selective inhibitors of human 11?-HSD1. Lead compounds display low nanomolar inhibition against human and mouse 11?-HSD1 and are selective with no activity against 11?-HSD2 and 17?-HSD1. Selected potent 11?-HSD1 inhibitors show moderate metabolic stability upon incubation with human liver microsomes and weak inhibition of human CYP450 enzymes.
Project description:Elevated levels of active glucocorticoids have been implicated in the development of several phenotypes of metabolic syndrome, such as type?2 diabetes and obesity. 11?-Hydroxysteroid dehydrogenase type?1 (11?-HSD1) catalyses the intracellular conversion of inactive cortisone to cortisol. Selective 11?-HSD1 inhibitors have shown beneficial effects in various conditions, including diabetes, dyslipidemia and obesity. A series of adamantyl ethanone pyridyl derivatives has been identified, providing potent and selective inhibitors of human 11?-HSD1. Lead compounds display low nanomolar inhibition against human and mouse 11?-HSD1 and are selective for this isoform, with no activity against 11?-HSD2 and 17?-HSD1. Structure-activity relationship studies reveal that an unsubstituted pyridine tethered to an adamantyl ethanone motif through an ether or sulfoxide linker provides a suitable pharmacophore for activity. The most potent inhibitors have IC?? values around 34-48?nM against human 11?-HSD1, display reasonable metabolic stability in human liver microsomes, and weak inhibition of key human CYP450 enzymes.
Project description:Oxysterols previously were considered intermediates of bile acid and steroid hormone biosynthetic pathways. However, recent research has emphasized the roles of oxysterols in essential physiologic processes and in various diseases. Despite these discoveries, the metabolic pathways leading to the different oxysterols are still largely unknown and the biosynthetic origin of several oxysterols remains unidentified. Earlier studies demonstrated that the glucocorticoid metabolizing enzymes, 11?-hydroxysteroid dehydrogenase (11?-HSD) types 1 and 2, interconvert 7-ketocholesterol (7kC) and 7?-hydroxycholesterol (7?OHC). We examined the role of 11?-HSDs in the enzymatic control of the intracellular availability of 7?,27-dihydroxycholesterol (7?27OHC), a retinoid-related orphan receptor ? (ROR?) ligand. We used microsomal preparations of cells expressing recombinant 11?-HSD1 and 11?-HSD2 to assess whether 7?27OHC and 7-keto,27-hydroxycholesterol (7k27OHC) are substrates of these enzymes. Binding of 7?27OHC and 7k27OHC to 11?-HSDs was studied by molecular modeling. To our knowledge, the stereospecific oxoreduction of 7k27OHC to 7?27OHC by human 11?-HSD1 and the reverse oxidation reaction of 7?27OHC to 7k27OHC by human 11?-HSD2 were demonstrated for the first time. Apparent enzyme affinities of 11?-HSDs for these novel substrates were equal to or higher than those of the glucocorticoids. This is supported by the fact that 7k27OHC and 7?27OHC are potent inhibitors of the 11?-HSD1-dependent oxoreduction of cortisone and the 11?-HSD2-dependent oxidation of cortisol, respectively. Furthermore, molecular docking calculations explained stereospecific enzyme activities. Finally, using an inducible ROR? reporter system, we showed that 11?-HSD1 and 11?-HSD2 controlled ROR? activity. These findings revealed a novel glucocorticoid-independent prereceptor regulation mechanism by 11?-HSDs that warrants further investigation.
Project description:Glucocorticoid (GC) hormones act on the brain to regulate diverse functions, from behavior and homeostasis to the activity of the hypothalamic-pituitary-adrenal axis. Local regeneration and metabolism of GCs can occur in target tissues through the actions of the 11?-hydroxysteroid dehydrogenases [11 beta-hydroxysteroid dehydrogenase type 1 (11?-HSD1) and 11 beta-hydroxysteroid dehydrogenase type 2 (11?-HSD2), respectively] to regulate access to GC receptors. Songbirds have become especially important model organisms for studies of stress hormone action; however, there has been little focus on neural GC metabolism. Therefore, we tested the hypothesis that 11?-HSD1 and 11?-HSD2 are expressed in GC-sensitive regions of the songbird brain. Localization of 11?-HSD expression in these regions could provide precise temporal and spatial control over GC actions. We quantified GC sensitivity in zebra finch (Taeniopygia guttata) brain by measuring glucocorticoid receptor (GR) and mineralocorticoid receptor (MR) expression across six regions, followed by quantification of 11?-HSD1 and 11?-HSD2 expression. We detected GR, MR, and 11?-HSD2 mRNA expression throughout the adult brain. Whereas 11?-HSD1 expression was undetectable in the adult brain, we detected low levels of expression in the brain of developing finches. Across several adult brain regions, expression of 11?-HSD2 covaried with GR and MR, with the exception of the cerebellum and hippocampus. It is possible that receptors in these latter two regions require direct access to systemic GC levels. Overall, these results suggest that 11?-HSD2 expression protects the adult songbird brain by rapid metabolism of GCs in a context and region-specific manner.
Project description:BACKGROUND: 11?-Hydroxysteroid dehydrogenase 1 (11?-HSD1) activates glucocorticoid locally in liver and fat tissues to aggravate metabolic syndrome. 11?-HSD1 selective inhibitor can be used to treat metabolic syndrome. Curcumin and its derivatives as selective inhibitors of 11?-HSD1 have not been reported. METHODOLOGY: Curcumin and its 12 derivatives were tested for their potencies of inhibitory effects on human and rat 11?-HSD1 with selectivity against 11?-HSD2. 200 mg/kg curcumin was gavaged to adult male Sprague-Dawley rats with high-fat-diet-induced metabolic syndrome for 2 months. RESULTS AND CONCLUSIONS: Curcumin exhibited inhibitory potency against human and rat 11?-HSD1 in intact cells with IC50 values of 2.29 and 5.79 µM, respectively, with selectivity against 11?-HSD2 (IC50, 14.56 and 11.92 µM). Curcumin was a competitive inhibitor of human and rat 11?-HSD1. Curcumin reduced serum glucose, cholesterol, triglyceride, low density lipoprotein levels in high-fat-diet-induced obese rats. Four curcumin derivatives had much higher potencies for Inhibition of 11?-HSD1. One of them is (1E,4E)-1,5-bis(thiophen-2-yl) penta-1,4-dien-3-one (compound 6), which had IC50 values of 93 and 184 nM for human and rat 11?-HSD1, respectively. Compound 6 did not inhibit human and rat kidney 11?-HSD2 at 100 µM. In conclusion, curcumin is effective for the treatment of metabolic syndrome and four novel curcumin derivatives had high potencies for inhibition of human 11?-HSD1 with selectivity against 11?-HSD2.
Project description:Reducing glucocorticoid exposure in the brain via intracellular inhibition of the cortisol-regenerating enzyme 11?-hydroxysteroid dehydrogenase type 1 (11?-HSD1) has emerged as a therapeutic strategy to treat cognitive impairment in early Alzheimer's disease (AD). We sought to discover novel, brain-penetrant 11?-HSD1 inhibitors as potential medicines for the treatment of AD.Medicinal chemistry optimization of a series of amido-thiophene analogues was performed to identify potent and selective 11?-HSD1 inhibitors with optimized oral pharmacokinetics able to access the brain. Single and multiple ascending dose studies were conducted in healthy human subjects to determine the safety, pharmacokinetic and pharmacodynamic characteristics of the candidate compound.UE2343 was identified as a potent, orally bioavailable, brain-penetrant 11?-HSD1 inhibitor and selected for clinical studies. No major safety issues occurred in human subjects. Plasma adrenocorticotropic hormone was elevated (a marker of systemic enzyme inhibition) at doses of 10 mg and above, but plasma cortisol levels were unchanged. Following multiple doses of UE2343, plasma levels were approximately dose proportional and the terminal t1/2 ranged from 10 to 14 h. The urinary tetrahydrocortisols/tetrahydrocortisone ratio was reduced at doses of 10 mg and above, indicating maximal 11?-HSD1 inhibition in the liver. Concentrations of UE2343 in the CSF were 33% of free plasma levels, and the peak concentration in CSF was ninefold greater than the UE2343 IC50 .UE2343 is safe, well tolerated and reaches the brain at concentrations predicted to inhibit 11?-HSD1. UE2343 is therefore a suitable candidate to test the hypothesis that 11?-HSD1 inhibition in brain improves memory in patients with AD.
Project description:17?-estradiol (E2), the most potent estrogen in humans, known to be involved in the development and progession of estrogen-dependent diseases (EDD) like breast cancer and endometriosis. 17?-HSD1, which catalyses the reduction of the weak estrogen estrone (E1) to E2, is often overexpressed in breast cancer and endometriotic tissues. An inhibition of 17?-HSD1 could selectively reduce the local E2-level thus allowing for a novel, targeted approach in the treatment of EDD. Continuing our search for new nonsteroidal 17?-HSD1 inhibitors, a novel pharmacophore model was derived from crystallographic data and used for the virtual screening of a small library of compounds. Subsequent experimental verification of the virtual hits led to the identification of the moderately active compound 5. Rigidification and further structure modifications resulted in the discovery of a novel class of 17?-HSD1 inhibitors bearing a benzothiazole-scaffold linked to a phenyl ring via keto- or amide-bridge. Their putative binding modes were investigated by correlating their biological data with features of the pharmacophore model. The most active keto-derivative 6 shows IC??-values in the nanomolar range for the transformation of E1 to E2 by 17?-HSD1, reasonable selectivity against 17?-HSD2 but pronounced affinity to the estrogen receptors (ERs). On the other hand, the best amide-derivative 21 shows only medium 17?-HSD1 inhibitory activity at the target enzyme as well as fair selectivity against 17?-HSD2 and ERs. The compounds 6 and 21 can be regarded as first benzothiazole-type 17?-HSD1 inhibitors for the development of potential therapeutics.
Project description:Impaired 11?-hydroxysteroid dehydrogenase type 2 (11?-HSD2)-dependent cortisol inactivation can lead to electrolyte dysbalance, hypertension and cardiometabolic disease. Furthermore, placental 11?-HSD2 essentially protects the fetus from high maternal glucocorticoid levels, and its impaired function has been associated with altered fetal growth and a higher risk for cardio-metabolic diseases in later life. Despite its important role, 11?-HSD2 is not included in current off-target screening approaches. To identify potential 11?-HSD inhibitors among approved drugs, a pharmacophore model was used for virtual screening, followed by biological assessment of selected hits. This led to the identification of several azole fungicides as 11?-HSD inhibitors, showing a significant structure-activity relationship between azole scaffold size, 11?-HSD enzyme selectivity and inhibitory potency. A hydrophobic linker connecting the azole ring to the other, more polar end of the molecule was observed to be favorable for 11?-HSD2 inhibition and selectivity over 11?-HSD1. The most potent 11?-HSD2 inhibition, using cell lysates expressing recombinant human 11?-HSD2, was obtained for itraconazole (IC50 139±14nM), its active metabolite hydroxyitraconazole (IC50 223±31nM) and posaconazole (IC50 460±98nM). Interestingly, experiments with mouse and rat kidney homogenates showed considerably lower inhibitory activity of these compounds towards 11?-HSD2, indicating important species-specific differences. Thus, 11?-HSD2 inhibition by these compounds is likely to be overlooked in preclinical rodent studies. Inhibition of placental 11?-HSD2 by these compounds, in addition to the known inhibition of cytochrome P450 enzymes and P-glycoprotein efflux transport, might contribute to elevated local cortisol levels, thereby affecting fetal programming.
Project description:Activity and selectivity assessment of new bi-aryl amide 11?-hydroxysteroid dehydrogenase 1 (11?-HSD1) inhibitors, prepared in a modular manner via Suzuki cross-coupling, are described. Several compounds inhibiting 11?-HSD1 at nanomolar concentrations were identified. Compounds 2b, 3e, 7b and 12e were shown to selectively inhibit 11?-HSD1 over 11?-HSD2, 17?-HSD1 and 17?-HSD2. These inhibitors also potently inhibited 11?-HSD1 activity in intact HEK-293 cells expressing the recombinant enzyme and in intact primary human keratinocytes expressing endogenous 11?-HSD1. Moreover, compounds 2b, 3e and 12e were tested for their activity in human skin biopsies. They were able to prevent, at least in part, both the cortisone- and the UV-mediated decreases in collagen content. Thus, inhibition of 11?-HSD1 by these compounds can be further investigated to delay or prevent UV-mediated skin damage and skin aging.