A Multitarget Approach against Neuroinflammation: Alkyl Substituted Coumarins as Inhibitors of Enzymes Involved in Neurodegeneration.

Neurodegenerative disorders (NDs) include a large range of diseases characterized by neural dysfunction with a multifactorial etiology. The most common NDs are Alzheimer's disease and Parkinson's disease, in which cholinergic and dopaminergic systems are impaired, respectively. Despite different brain regions being affected, oxidative stress and inflammation were found to be common triggers in the pathogenesis and progression of both diseases. By taking advantage of a multi-target approach, in this work we explored alkyl substituted coumarins as neuroprotective agents, capable to reduce oxidative stress and inflammation by inhibiting enzymes involved in neurodegeneration, among which are Carbonic Anhydrases (CAs), Monoamine Oxidases (MAOs), and Cholinesterases (ChEs). The compounds were synthesized and profiled against the three targeted enzymes. The binding mode of the most promising compounds (7 and 9) within MAO-A and -B was analyzed through molecular modeling studies, providing and explanation for the different selectivities observed for the MAO isoforms. In vitro biological studies using LPS-stimulated rat astrocytes showed that some compounds were able to counteract the oxidative stress-induced neuroinflammation and hamper interleukin-6 secretion, confirming the success of this multitarget approach.

Kinetic Study on the Base-Catalyzed Imine-Enamine Tautomerism of a Chiral Biologically Active Isoxazoline Derivative by HPLC on Amylose Tris(3,5-dimethylphenylcarbamate) Chiral Stationary Phase.

Isoxazoline is a nitrogen- and oxygen-containing five-membered heterocyclic scaffold with diverse biological profiles such as antimicrobial, fungicidal, anticancer, antiviral, analgesic and anti-inflammatory activity. Accordingly, the use of this peculiar structural framework in drug discovery is a successful strategy for the development of new drug candidates. Here, a chiral saccharin/isoxazoline hybrid was considered to investigate the tendency of the imine moiety of the heterocyclic ring to tautomerize to the enamine form in the presence of a basic catalyst. The pseudo-first-order rate constants for the base-catalyzed tautomerization process were measured in different solvents and at different temperatures by off-column kinetic experiments based on the amylose (3,5-dimethylphenylcarbamate)-type chiral stationary phase. The kinetic results obtained in this study may be a useful aid in the perspective of designing experimental conditions to control the stereointegrity of these types of pharmacologically active compounds and drive their synthesis toward the preferred, imine or enamine, tautomer.

Novel therapeutic opportunities for Toxoplasma gondii, Trichomonas vaginalis, and Giardia intestinalis infections.

INTRODUCTION: Toxoplasma gondii, Trichomonas vaginalis, and Giardia intestinalis are the causative agents of toxoplasmosis, trichomoniasis, and giardiasis, three important infections threatening human health and affecting millions of people worldwide. Although drugs and treatment are available to fight these protozoan parasites, side effects and increasing drug resistance require continuous efforts for the development of novel effective drugs. AREAS COVERED: The patents search was carried out in September/October 2022 with four official scientific databases (Espacenet, Scifinder, Reaxys, Google Patents). Treatments for toxoplasmosis, trichomoniasis, and giardiasis (2015-2022) have been grouped according to their chemotypes. In particular, novel chemical entities have been reported and investigated for their structure-activity relationship, when accessible. On the other hand, drug repurposing, extensively exploited to obtain novel antiprotozoal treatment, has been in-depth described. Finally, natural metabolites and extracts have also been reported. EXPERT OPINION: T. gondii, T. vaginalis, and G. intestinalis are protozoan infections usually controlled by immune system in immunocompetent patients; however, they could represent a threatening health for immunocompromised people. The needs of novel effective drugs, endowed with new mechanisms of actions, arises from the increasing drug resistance affecting antibiotic as well as antiprotozoal therapies. In this review different therapeutic approaches to treat protozoan infections have been reported.

Color stability of resin-based composites exposed to smoke. A systematic review.

OBJECTIVE: To conduct a systematic review on color stability of dental resin-based composites (RBC) exposed to conventional and electronic cigarettes. MATERIALS AND METHODS: In vitro studies reporting on the color stability of RBC exposed to conventional cigarettes or to e-cigarettes: both Tobacco Heating Systems (THS) and Electronic Nicotine Delivery Systems (ENDS). The quality of the included studies was assessed with the QUIN tool (risk-of-bias tool for assessing in vitro studies conducted in dentistry). A systematic search, following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) was performed on four (n = 4) databases (Embase, Medline, Scopus, Web of Science) for articles published until March 28th, 2022. RESULTS: Of the 365 screened articles, 13 were included in this review. All the included articles analyzed conventional cigarette smoke (CS), four analyzed Electronic Nicotine Delivery Systems (ENDS) and two Tobacco Heating Systems (THS). In terms of study design, smoke exposure time, smoke flow, type and number of cigarettes a high variability was reported. CONCLUSIONS: The available evidence suggests that CS smoke significantly affects color stability. Electronic cigarettes show less color change that seems to be easily recovered under clinical acceptability thresholds, although evidence is scarce. CLINICAL SIGNIFICANCE: Clinicians should be aware, and should therefore warn their patients, that RBCs are subjected to irreversible color change if exposed to smoke. Electronic cigarettes (both ENDS and THS) induce less color change that can be recovered with repolishing or whitening procedures.

Novel Insights on CAI-CORM Hybrids: Evaluation of the CO Releasing Properties and Pain-Relieving Activity of Differently Substituted Coumarins for the Treatment of Rheumatoid Arthritis.

Pain control is among the most important healthcare services in patients affected by rheumatoid arthritis (RA), but the current therapeutic options (i.e., disease-modifying anti-rheumatic drugs) are limited by the risk of the side effects. In this context, we proposed an innovative approach based on the hybridization between carbonic anhydrase inhibitors (CAIs) and CO releasing molecules (CORMs). The resulting CAI-CORM hybrids were revealed to possess strong anti-inflammatory effects in in vitro models of diseases and to relieve ache symptoms in an in vivo RA rat model. In this work, we have deepened the study of these promising hybrids, designing a library of coumarin-based compounds, also including internal dicobalt hexacarbonyl systems. The results obtained from the CO releasing study, the CA inhibitory activity, and the in vivo pain-relief efficacy evaluation in the RA rat model confirmed the success of this strategy, allowing us to consider CAI-CORM hybrids promising anti-nociceptive agents against arthritis.

Metal-organic frameworks (MOFs) as biomolecules drug delivery systems for anticancer purposes.

Metal-organic frameworks (MOF) have been studied for infinite applications. Among these, anticancer therapy has surely attracted great interest due to the intrinsic characteristics of MOFs: large surface area, tuneable porosity, remarkable biocompatibility, easy production, and the possibility to further functionalize the realized nanoparticles are the main reasons that make MOFs the ideal candidates to overcome traditional chemotherapy limits and resistance. Smart MOFs are becoming particularly relevant, as they can be activated by specific endogenous or exogenous stimuli and release their cargo only under the selected conditions. Tumor microenvironment (TME) offers the possibility to take advantage of its peculiar composition to design and build smart nanoparticles, able to selectively release the therapeutic payload only in the environment surrounding cancer cells or directly in the intracellular environment. In this review, we have summarized novel and innovative works describing anticancer MOF-based nanoparticles loaded with biomolecules published in the last three years. The reported papers have been selected with special focus on TME-responsive MOFs and the synthetic procedures employed by research groups have been reported for almost all works, to further underline the myriad of possibilities offered by these hybrid metal-organic structures.

In vitro procedures for color stability evaluation of dental resin-based composites exposed to smoke: A scoping review.

The objective of this study was to review the experimental in vitro procedures employed to assess the color stability of resin-based composites exposed to smoke. A literature search was performed on four databases (Pubmed, Scopus, Embase, and Web of Science). The quality of the included papers was assessed with the Cochrane risk-of-bias tool (RoB 2). In total, 25 studies were selected for full-text reading, from which 12 were included in the review. The assessed variables were: dimensions, shape, time before exposure, finishing, smoke-type, exposure time, brushing simulation, color, measuring system, repolishing. A disk-shaped specimen was used in most studies (n=10) with a 2 mm thickness (n=7) in custom made devices equipped with vacuum pumps or in Vitrocell systems. Spectrophotometers were the most used devices (n=11). Color differences were quantified with ΔE (n=12), ΔE00 (n=1). The large variety of staining procedures suggests the need for standardization.

Novel Insights on Human Carbonic Anhydrase Inhibitors Based on Coumalic Acid: Design, Synthesis, Molecular Modeling Investigation, and Biological Studies.

Human carbonic anhydrase (hCA, EC 4.2.1.1) isoforms IX and XII are overexpressed in solid hypoxic tumors, and they are considered as prognostic tools and therapeutic targets for cancer. Based on a molecular simplification of the well-known coumarin scaffold, we developed a new series of derivatives of the pyran-2-one core. The new compounds are endowed with potent and selective inhibitory activity against the tumor-related hCA isoforms IX and XII, in the low nanomolar range, whereas they are inactive against the two cytosolic off-targets hCA I and II. The compounds exhibiting the best hCA inhibition were further investigated against the breast adenocarcinoma cell line (MCF7) in hypoxic conditions, evaluating their ability to eventually synergize with doxorubicin. The compounds' biocompatibility on healthy cells was also tested and confirmed on Human Gingival Fibroblasts (HGFs). Furthermore, the possible binding mode of all compounds to the active site of the tumor-associated human CA IX was investigated by computational techniques which predicted the binding conformations and the persistency of binding poses within the active site of the enzyme, furnishing relevant data for the design of tight binding inhibitors.

An updated patent review on monoamine oxidase (MAO) inhibitors.

INTRODUCTION: Monoamine oxidase (MAO) inhibitors are currently used as antidepressants (selective MAO-A inhibitors) or as co-adjuvants for neurodegenerative diseases (selective MAO-B inhibitors). The research within this field is attracting attention due to their crucial role in the modulation of brain functions, mood, and cognitive activity, and monoamine catabolism. AREAS COVERED: MAO inhibitors (2018-2021) are discussed according to their chemotypes. Structure-activity relationships are derived for each chemical scaffold (propargylamines, chalcones, indoles, benzimidazoles, (iso)coumarins, (iso)benzofurans, xanthones, and tetralones), while the chemical entities were divided into newly synthesized molecules and natural metabolites. The mechanism of action and type of inhibition are also considered. Lastly, new therapeutic applications are reported, which demonstrates the clinical potential of these inhibitors as well as the possibility of repurposing existing drugs for a variety of diseases. EXPERT OPINION: MAO inhibitors here reported exhibit different potencies and isoform selectivity. These compounds are clinically licensed for multi-faceted neurodegenerative pathologies due to their ability to also act against other relevant targets (cholinesterases, inflammation, and oxidative stress). Moreover, the drug repurposing approach is an attractive strategy by which MAO inhibitors may be applied for the treatment of prostate cancer, inflammation, vertigo, and type 1 diabetes.

An innovative spectroscopic approach for qualitative and quantitative evaluation of Mb-CO from myoglobin carbonylation reaction through chemometrics methods.

In this work, an innovative approach using K-means and multivariate curve resolution-purity based algorithm (MCR-Purity) for the evaluation and quantification of carboxymyoglobin (Mb-CO) formation from Deoxy-Myoglobin (Deoxy-Mb) was presented. Through a multilevel multifactor experimental design, samples with different concentrations of Mb-CO were created. The UV-Vis spectra of these samples were submitted to K-means analysis, finding 3 clusters. The mean spectra of the clusters were extracted and it was possible to detect 2 totally differentiable groups through peaks 423 and 434 nm, which are wavelengths related to the Mb-CO and Deoxy-Mb components, respectively. The spectral data were subjected to MCR-Purity analysis. The MCR-Purity result successfully described the analyzed reaction, explaining more than 99.9% of the variance (R2) with a LOF of 1.43%. Then, a predictive model of MbCO was created through the linear relationship between MCR-Purity contributions and known concentrations of MbCO. The performance parameters of the created predictive model were R2CV = 0.98, RMSECV = 0.58 and RPDcv = 7.8 for the training set, and R2P = 0.98, RMSEP = 0.7 and RPDp = 6.8 for the test set. Thus, the predictive model presented an excellent performance considering that the Mb-CO variation is comprised between 0 and 21 µM. Therefore, these results demonstrate that the application of the proposed strategy to the analysis of spectral data presenting overlapping bands is feasible and robust.

Synthesis and Evaluation of Thymol-Based Synthetic Derivatives as Dual-Action Inhibitors against Different Strains of H. pylori and AGS Cell Line.

Following a similar approach on carvacrol-based derivatives, we investigated the synthesis and the microbiological screening against eight strains of H. pylori, and the cytotoxic activity against human gastric adenocarcinoma (AGS) cells of a new series of ether compounds based on the structure of thymol. Structural analysis comprehended elemental analysis and 1H/13C/19F NMR spectra. The analysis of structure-activity relationships within this molecular library of 38 structurally-related compounds reported that some chemical modifications of the OH group of thymol led to broad-spectrum growth inhibition on all isolates. Preferred substitutions were benzyl groups compared to alkyl chains, and the specific presence of functional groups at para position of the benzyl moiety such as 4-CN and 4-Ph endowed the most anti-H. pylori activity toward all the strains with minimum inhibitory concentration (MIC) values up to 4 µg/mL. Poly-substitution on the benzyl ring was not essential. Moreover, several compounds characterized by the lowest minimum inhibitory concentration/minimum bactericidal concentration (MIC/MBC) values against H. pylori were also tested in order to verify a cytotoxic effect against AGS cells with respect to 5-fluorouracil and carvacrol. Three derivatives can be considered as new lead compounds alternative to current therapy to manage H. pylori infection, preventing the occurrence of severe gastric diseases. The present work confirms the possibility to use natural compounds as templates for the medicinal semi-synthesis.

Dual Carbonic Anhydrase IX/XII Inhibitors and Carbon Monoxide Releasing Molecules Modulate LPS-Mediated Inflammation in Mouse Macrophages.

Low concentrations of carbon monoxide (CO) were reported to exhibit anti-inflammatory effects when administered in cells by suitable chemotypes such as CO releasing molecules (CO-RMs). In addition, the pH-modulating abilities of specific carbonic anhydrase isoforms played a crucial role in different models of inflammation and neuropathic pain. Herein, we report a series of chemical hybrids consisting of a Carbonic Anhydrase (CA) inhibitor linked to a CO-RM tail (CAI/CO-RMs). All compounds and their precursors were first tested in vitro for their inhibition activity against the human CA I, II, IX, and XII isoforms as well their CO releasing properties, aiming at corroborating the data by means of molecular modelling techniques. Then, their impact on metabolic activity modulation of RAW 264.7 mouse macrophages for 24 and 48 h was assessed with or without lipopolysaccharide (LPS) stimulation. The compounds were shown to counteract the inflammatory stimulus as also indicated by the reduced tumor necrosis factor alpha (TNF-α) release after treatment. All the biological results were compared to those of N-acetylcysteine (NAC) as a reference antioxidant compound. Within the series, two CAI/CO-RM hybrids (1 and 2), bearing both the well-known scaffold able to inhibit CAs (acesulfame) and the cobalt-based CO releasing portion, induced a higher anti-inflammatory effect up to 48 h at concentrations lower than NAC.

New Aspects of Monoamine Oxidase B Inhibitors: The Key Role of Halogens to Open the Golden Door.

A large plethora of drugs and promising lead compounds contain halogens in their structures. The introduction of such moieties strongly modulates their physical-chemical features as well as pharmacokinetic and pharmacodynamic profile. The most important outcome was shown to be the ability of these halogens to favourably influence the drug-target interaction and energetic stability within the active site by the establishment of halogen bonds. This review attempted to demonstrate the key role exerted by these versatile moieties when correctly located in an organic scaffold to display Monoamine Oxidase (MAO) inhibition and selectivity towards the B isoform of this important enzyme. Human MAOs are well-recognized as therapeutic targets for mood disorders and neurodegenerative diseases and medicinal chemists were prompted to discover the structural requirements crucial to discriminate the slight differences between the active sits of the two isoforms (MAO-A and MAOB). The analysis of the structure-activity relationships of the most important scaffolds (hydrazothiazoles, coumarins, chromones, chalcones, pyrazolines) and the impact of halogen (F, Cl, Br and I) insertion on this biological activity and isozyme selectivity have been reported being a source of inspiration for the medicinal chemists.

Synthesis and Biological Evaluation of Carvacrol-Based Derivatives as Dual Inhibitors of H. pylori Strains and AGS Cell Proliferation.

This study reports on the synthesis, structural assessment, microbiological screening against several strains of H. pylori and antiproliferative activity against human gastric adenocarcinoma (AGS) cells of a large series of carvacrol-based compounds. Structural analyses consisted of elemental analysis, 1H/13C/19F NMR spectra and crystallographic studies. The structure-activity relationships evidenced that among ether derivatives the substitution with specific electron-withdrawing groups (CF3 and NO2) especially in the para position of the benzyl ring led to an improvement of the antimicrobial activity, whereas electron-donating groups on the benzyl ring and ethereal alkyl chains were not tolerated with respect to the parent compound (MIC/MBC = 64/64 µg/mL). Ester derivatives (coumarin-carvacrol hybrids) displayed a slight enhancement of the inhibitory activity up to MIC values of 8-16 µg/mL. The most interesting compounds exhibiting the lowest MIC/MBC activity against H. pylori (among others, compounds 16 and 39 endowed with MIC/MBC values ranging between 2/2 to 32/32 µg/mL against all the evaluated strains) were also assayed for their ability to reduce AGS cell growth with respect to 5-Fluorouracil. Some derivatives can be regarded as new lead compounds able to reduce H. pylori growth and to counteract the proliferation of AGS cells, both contributing to the occurrence of gastric cancer.

Natural compounds and extracts as novel antimicrobial agents.

Introduction: Antimicrobial resistance is a worldwide problem accounting for the reduction or in some cases absence of drugs effectiveness normally used in infections treatment. In the light of the even more spread ability of microbials to develop resistance, there is an urgent necessity to find novel and alternative routes to fight infections. Natural compounds or extracts can be a valid alternative either as monotherapy or as adjuvant in order to improve the effectiveness of the failing drugs. Areas covered: This review provides a comprehensive update (2018-2020) on the development state of innovative antimicrobial agents based on natural compounds and extracts, also describing their compositions, methods of production and use, mechanism of action, along with anti-microbial data when available. Expert opinion: Owing to the pivotal role that natural compounds often cover in the finding of novel drugs, their in-depth analysis could pave the way to the discovery of new antimicrobial agents. Most of the alternative approaches reported in this short review were validated through in vitro and in vivo (animal as well as human) models. The employment of natural derived compounds and extracts, alone or in combination with classical antimicrobial drugs, as antimicrobial agents could represent an important achievement to challenge pathogens resistant mechanisms.

Novel insights on saccharin- and acesulfame-based carbonic anhydrase inhibitors: design, synthesis, modelling investigations and biological activity evaluation.

A large library of saccharin and acesulfame derivatives has been synthesised and evaluated against four isoforms of human carbonic anhydrase, the two off-targets hCA I/II and the tumour related isoforms hCA IX/XII. Different strategies of scaffold modification have been attempted on both saccharin as well as acesulfame core leading to the obtainment of 60 compounds. Some of them exhibited inhibitory activity in the nanomolar range, albeit some of the performed changes led to either micromolar activity or to its absence, against hCA IX/XII. Molecular modelling studies focused the attention on the binding mode of these compounds to the enzyme. The proposed inhibition mechanism is the anchoring to zinc-bound water molecule. Docking studies along with molecular dynamics also underlined the importance of the compounds flexibility (e.g. achieved through the insertion of methylene group) which favoured potent and selective hCA inhibition.

Chalcones: Unearthing their therapeutic possibility as monoamine oxidase B inhibitors.

In the last years the continuous efforts in the development of novel and effective inhibitors of human monoamine oxidases (hMAOs) promoted the discovery of new agents able to effectively and selectively bound one of the two isoforms (hMAO-A and hMAO-B). However, the parent chalcone scaffold still covers an important role in hMAOs inhibition. In the present work, we focused our attention on the researches performed in the last five years, involving chalcones or compounds that can be correlated to them. We classified the chalcones into different groups depending on their structural characteristics or common molecular properties. In this regard, we also considered chalcones based on heterocycles and compounds endowed with scaffolds containing a masked chalcone motif. When structural attributes could not be used, we took advantage of enzymatic activity to arrange compounds in a group. We followed this approach for the multitarget agents. Finally, we also analysed the naturally occurring chalcones. All the sections were discussed exhaustively and the structure-activity relationship (SAR) analyses were sustained by means of detailed images describing the effects related to the substituents or structural changes.

Synthesis and evaluation of a large library of nitroxoline derivatives as pancreatic cancer antiproliferative agents.

Pancreatic cancer (PC) is one of the deadliest carcinomas and in most cases, which are diagnosed with locally advanced or metastatic disease, current therapeutic options are highly unsatisfactory. Based on the anti-proliferative effects shown by nitroxoline, an old urinary antibacterial agent, we explored a large library of newly synthesised derivatives to unravel the importance of the OH moiety and pyridine ring of the parent compound. The new derivatives showed a valuable anti-proliferative effect and some displayed a greater effect as compared to nitroxoline against three pancreatic cancer cell lines with different genetic profiles. In particular, in silico pharmacokinetic data, clonogenicity assays and selectivity indexes of the most promising compounds showed several advantages for such derivatives, as compared to nitroxoline. Moreover, some of these novel compounds had stronger effects on cell viability and/or clonogenic capacity in PC cells as compared to erlotinib, a targeted agent approved for PC treatment.

1,3-Dipolar Cycloaddition, HPLC Enantioseparation, and Docking Studies of Saccharin/Isoxazole and Saccharin/Isoxazoline Derivatives as Selective Carbonic Anhydrase IX and XII Inhibitors.

Two series of saccharin/isoxazole and saccharin/isoxazoline hybrids were synthesized by 1,3-dipolar cycloaddition. The new compounds showed to be endowed with potent and selective inhibitory activity against the cancer-related human carbonic anhydrase (hCA) IX and XII isoforms in the nanomolar range, while no affinity was encountered for off-targets, such as hCA I and II. Successive enantioseparation on a milligram scale of the most representative compounds led to the discovery that (S)-isomers were more potent than their corresponding (R)-enantiomers. Lastly, molecular modeling studies were conducted to define those structural requirements that were responsible for the discrimination among selected human isoforms of carbonic anhydrases. Two nanomolar hCA IX and XII inhibitors were also screened for their selective toxicity against non tumoral primary cells (fibroblasts) and against a breast adenocarcinoma cell line (MCF7) in hypoxic environment. The efficacious combination of these compounds with doxorubicin on MCF7 cells was demonstrated after 72 h of treatment.

High-performance liquid chromatography enantioseparation of chiral 2-(benzylsulfinyl)benzamide derivatives on cellulose tris(3,5-dichlorophenylcarbamate) chiral stationary phase.

Recently it has been reported that immobilized chlorinated-type chiral stationary phases based on cellulose tris(3,5-dichlorophenylcarbamate) are able to express an outstanding enantioselectivity towards the structure of 2-(benzylsulfinyl)benzamide. We now introduce two homologue series of chiral sulfoxides based on the same 2-(sulfinyl)benzoyl core as the prototype of new selectands for HPLC, whose enantioselectivity could be modulable through the replacement of the benzyl group with an unbranched alkyl chain varying in length from 1 to 5 carbon atoms. HPLC parameters such as mobile phase composition and column temperature have been carefully evaluated in order to get pertinent structure-enantioselectivity relationships. The enantiomer elution order was unambiguously determined by a combined strategy involving theoretical and experimental procedures. Two cases of temperature-dependent inversion of the elution order of enantiomers in the operative temperature range of chiral chromatographic support were observed.