The biosynthesis of the mineralocorticoid hormone aldosterone involves a multistep hydroxylation of 11‐deoxycorticosterone at the 11‐ and 18‐positions, resulting in the formation of corticosterone and 18‐hydroxycorticosterone, the final precursor of aldosterone. Two members of the cytochrome P450 11B family, CYP11B1 and CYP11B2, are known to catalyze these 11‐ and 18‐hydroxylations, however, only CYP11B2 can oxidize 18‐hydroxycorticosterone to aldosterone. It is unknown what sequence of hydroxylations leads to the formation of 18‐hydroxycorticosterone. In this study we have investigated which of the possible conversion paths towards formation of 18‐hydroxycorticosterone and aldosterone are most likely from the ligand perspective. Therefore, we combined quantum mechanical investigations on the steroid conformations of 11‐deoxycorticosterone and its ensuing reaction intermediates with Fukui indices calculations to predict the reactivity of their carbon atoms for an attack by the iron‐oxygen species. Both F − and F 0 were calculated to account for different mechanisms of substrate conversion. We show which particular initial conformations of 11‐deoxycorticosterone and which conversion paths are likely to result in the successful synthesis of aldosterone, and thereby may be representative for the mechanism of aldosterone biosynthesis by CYP11B2. Moreover, we found that the most likely path for aldosterone synthesis coincides with the substrate conformation proposed in an earlier publication (Ref. 2 ). To summarize, we show that on a theoretical and strictly ligand‐directed basis only a limited number of reaction paths in the conversion of 11‐deoxycorticosterone to aldosterone is possible. Despite its theoretical nature, this knowledge may help to understand the catalytic function of CYP11B1 and CYP11B2. © 2011 Wiley Periodicals, Inc. J Comput Chem, 2011

Background— The molecular pathways that control the wound healing after myocardial infarction (MI) are not completely elucidated. One of these pathways is the Wnt/Frizzled pathway. In this study, we evaluated Frizzled as a novel therapeutic target for MI. These Frizzled proteins act as receptors for Wnt proteins and were previously shown to be expressed in the healing infarct. Methods and Results— Wnt/Frizzled signaling has been studied for decades, but synthetic ligands that interfere with the interaction between Wnts and Frizzled have not been described to date. Here we report the selection of 3 peptides derived from regions of high homology between Wnt3a and Wnt5a that act as antagonists for Frizzled proteins. UM206, the peptide with the highest affinity, antagonized the effect of Wnt3a and Wnt5a in different in vitro assays. Administration of UM206 to mice for 5 weeks, starting immediately after the induction of MI, reduced infarct expansion and increased the numbers of capillaries and myofibroblasts in the infarct area. Moreover, heart failure development was inhibited by this therapy. Conclusions— Blocking of Frizzled signaling reduces infarct expansion and preserves cardiac function after MI. Our findings underscore the potential of Frizzled receptors as a target for pharmacotherapy of cardiac remodeling after MI.

AIMS The Wnt/frizzled (Fzd) signal transduction cascade has been implicated in the proliferation, differentiation, and migration of many cell types, but the role of this pathway in cardiac fibroblast differentiation is not known. Our lab previously showed an up-regulation of Fzd-1 and -2 expression in myofibroblasts after myocardial infarction (MI), indicating a potential role for the Fzd receptor in fibroblast-myofibroblast differentiation. The present study was performed to further define the role of specific Wnt and Fzd proteins in the proliferation, migration, and differentiation of cardiac fibroblasts. METHODS AND RESULTS Because primary fibroblasts become senescent after a few passages and are difficult to transfect, we immortalized rat cardiac fibroblasts with telomerase [cardiac fibroblasts immortalized with telomerase (CFIT)]. Proliferation of CFIT was not significantly influenced by Wnt/Fzd signalling. The migration, however, was attenuated by all Wnt/Fzd combinations tested. Also, specific Wnt/Fzd combinations modulated the expression of the following myofibroblast markers: collagen Ialpha1, collagen III, fibronectin and its splice variants, and alpha-smooth muscle actin. CONCLUSION The results indicate that myofibroblast migration and differentiation, but not proliferation, can be modulated by interventions in Wnt/Fzd signalling. Therefore, Wnt/Fzd signalling may serve as a novel therapeutic target to ameliorate wound healing after MI.

Reducing aldosterone action is beneficial in various major diseases such as heart failure. Currently, this is achieved with mineralocorticoid receptor antagonists, however, aldosterone synthase (CYP11B2) inhibitors may offer a promising alternative. In this study, we used three-dimensional modeling of CYP11B2 to model the binding modes of the natural substrate 18-hydroxycorticosterone and the recently published CYP11B2 inhibitor R-fadrozole as a rational guide to design 44 structurally simple and achiral 1-benzyl-1H-imidazoles. Their syntheses, in vitro inhibitor potencies, and in silico docking are described. Some promising CYP11B2 inhibitors were identified, with our novel lead MOERAS115 (4-((5-phenyl-1H-imidazol-1-yl)methyl)benzonitrile) displaying an IC(50) for CYP11B2 of 1.7 nM, and a CYP11B2 (versus CYP11B1) selectivity of 16.5, comparable to R-fadrozole (IC(50) for CYP11B2 6.0 nM, selectivity 19.8). Molecular docking of the inhibitors in the models enabled us to generate posthoc hypotheses on their binding modes, providing a valuable basis for future studies and further design of CYP11B2 inhibitors.

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Aldosterone is synthesised by aldosterone synthase (CYP11B2). CYP11B2 has a highly homologous isoform, steroid 11β-hydroxylase (CYP11B1), which is responsible for the biosynthesis of aldosterone precursors and glucocorticoids. To investigate aldosterone biosynthesis and facilitate the search for selective CYP11B2 inhibitors, we constructed three-dimensional models for CYP11B1 and CYP11B2 for both human and rat. The models were constructed based on the crystal structure of Pseudomonas Putida CYP101 and Oryctolagus Cuniculus CYP2C5 . Small steric active site differences between the isoforms were found to be the most important determinants for the regioselective steroid synthesis. A possible explanation for these steric differences for the selective synthesis of aldosterone by CYP11B2 is presented. The activities of the known CYP11B inhibitors metyrapone, R -etomidate, R -fadrazole and S -fadrazole were determined using assays of V79MZ cells that express human CYP11B1 and CYP11B2, respectively. By investigating the inhibitors in the human CYP11B models using molecular docking and molecular dynamics simulations we were able to predict a similar trend in potency for the inhibitors as found in the in vitro assays. Importantly, based on the docking and dynamics simulations it is possible to understand the enantioselectivity of the human enzymes for the inhibitor fadrazole, the R -enantiomer being selective for CYP11B2 and the S -enantiomer being selective for CYP11B1.

Reversal of cardiac fibrosis is a major determinant of the salutary effects of mineralocorticoid receptor antagonists in heart failure. Recently, R-fadrozole was coined as an aldosterone biosynthesis inhibitor, offering an appealing alternative to mineralocorticoid receptor antagonists to block aldosterone action. The present study aimed to evaluate the effects of R- and S-fadrozole on plasma aldosterone and urinary aldosterone excretion rate and to compare their effectiveness vs. the mineralocorticoid receptor antagonist potassium canrenoate to reverse established cardiac fibrosis. Male lean spontaneously hypertensive heart failure (SHHF) rats (40 wk) were treated for 8 wk by sc infusions of low (0.24 mg/kg.d) or high (1.2 mg/kg.d) doses of R- or S-fadrozole or by potassium canrenoate via drinking water (7.5 mg/kg.d). At the high dose, plasma aldosterone levels were decreased similarly by R- and S-fadrozole, whereas urinary aldosterone excretion rate was reduced only by S-fadrozole. In contrast, whereas at the high dose, R-fadrozole effectively reversed preexistent left ventricular interstitial fibrosis by 50% (vs. 42% for canrenoate), S-fadrozole was devoid of an antifibrotic effect. The low doses of the fadrozole enantiomers did not change cardiac fibrosis or plasma aldosterone but similarly reduced urinary aldosterone excretion rate. In conclusion, R-fadrozole may possess considerable therapeutic merit because of its potent antifibrotic actions in the heart. However, the observed discordance between the aldosterone-lowering and antifibrotic effects of the fadrozole enantiomers raises some doubt about the mechanism by which R-fadrozole diminishes cardiac collagen and about the generality of the concept of lowering aldosterone levels to treat the diseased heart.

Immobilized 4-iodoimidazole 2 was used in a metal/halogen exchange reaction followed by treatment with electrophiles and subsequent cleavage from the resin to yield 4-substituted imidazoles 8-11. Grignard reaction with the resin-bound ketones 5 yielded the corresponding alcohols 11. This approach was used for a library synthesis of 35 imidazoles.

AbstractChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 100 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a “Full Text” option. The original article is trackable via the “References” option.

This Report reviews literature published Nov 1996-Dec 97; it is the third in a series that summarises organic reactions performed on solid supports and is designed as an update to Tetrahedron Reports no. 394 and 418. The reactions are presented in a graphical abstract format and examples are given of the following reactions: amide formation, aromatic substitution (nucleophilic/electrophilic), cleavage, condensation, cycloaddition, deprotection/protection, Grignard and related reactions, heterocycle formation, immobilisation onto resin, Michael addition, miscellaneous, multiple component condensation, olefin formation, oxidation, reduction and substitution (nucleophilic/electrophilic). Solid-phase peptide or other oligomer synthesis and reactions using soluble polymeric supports are not included. This report ends with the discussion of some trends in the domain under consideration. (C) 1998 Elsevier Science Ltd. All rights reserved.

This Report reviews literature published Nov 1995–Nov 1996 and summarises organic reactions performed on solid-supports. The reactions are presented in a graphical abstract format and examples of amide formation, aromatic nucleophilic substitution, cleavage, condensation, cycloaddition, deprotection, Grignard, heterocycle formation, immobilisation to resin, Michael addition, miscellaneous, multiple component condensation, olefin formation, oxidation, reduction and substitution (nucleophilic/electrophilic) are given. The review is designed as an update to Tetrahedron Report no. 394.

This Report reviews literature published Nov 1995-Nov 1996 and summarises organic reactions performed on solid-supports. The reactions are presented in a graphical abstract format and examples of amide formation, aromatic nucleophilic substitution, cleavage, condensation, cycloaddition, deprotection, Grignard, heterocycle formation, immobilisation to resin, Michael addition, miscellaneous, multiple component condensation, olefin formation, oxidation, reduction and substitution (nucleophilic/electrophilic) are given. The review is designed as an update to Tetrahedron Report no. 394. (C) 1997 Elsevier Science Ltd.

ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 100 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a “Full Text” option. The original article is trackable via the “References” option.

The antipsychotics currently used need to be improved, both in terms of efficacy and side-effect profile. In particular, the treatment of negative symptoms of schizophrenia and the occurrence of extrapyramidal side-effects need to be addressed. The high affinity for the 5-HT2A receptor of the atypical antipsychotic clozapine has renewed the interest in antagonists for this receptor. Several lines of evidence point towards a possible role of 5-HT2A receptors in the pathophysiology and treatment of schizophrenia. However, as the cause of schizophrenia is unknown the success of any approach based upon a specific biochemical mechanism can only be established in the clinic. Therefore, the real significance of 5-HT2A receptor antagonists for antipsychotic treatment will only become clear when data become available on the clinical efficacy of selective 5-HT2A receptor antagonists.In this manuscript the highlights published in the past decade are reviewed. Clinical and experimental data which support the rationale for 5-HT2A antagonists are listed. Next, the 5-HT2 receptor subtypes and regulation and structure of the 5-HT2A receptor are described. The 5-HT2A antagonists on the market or under (pre)clinical development as antipsychotics are listed. Finally, the major chemical classes of 5-HT2A receptor antagonists with relevance to psychosis are discussed.