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Prof. Giuliana Vinci
Department of Management, Sapienza University of Rome, 00185 Rome, Italy

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Review
Published: 05 August 2021 in Cancers
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Pancreatic cancer (PC) is one of the deadliest malignancies worldwide, since patients rarely display symptoms until an advanced and unresectable stage of the disease. Current chemotherapy options are unsatisfactory and there is an urgent need for more effective and less toxic drugs to improve the dismal PC therapy. Repurposing of non-oncology drugs in PC treatment represents a very promising therapeutic option and different compounds are currently being considered as candidates for repurposing in the treatment of this tumor. In this review, we provide an update on some of the most promising FDA-approved, non-oncology, repurposed drug candidates that show prominent clinical and preclinical data in pancreatic cancer. We also focus on proposed mechanisms of action and known molecular targets that they modulate in PC. Furthermore, we provide an explorative bioinformatic analysis, which suggests that some of the PC repurposed drug candidates have additional, unexplored, oncology-relevant targets. Finally, we discuss recent developments regarding the immunomodulatory role displayed by some of these drugs, which may expand their potential application in synergy with approved anticancer immunomodulatory agents that are mostly ineffective as single agents in PC.

ACS Style

Laura De Lellis; Serena Veschi; Nicola Tinari; Zhirajr Mokini; Simone Carradori; Davide Brocco; Rosalba Florio; Antonino Grassadonia; Alessandro Cama. Drug Repurposing, an Attractive Strategy in Pancreatic Cancer Treatment: Preclinical and Clinical Updates. Cancers 2021, 13, 3946 .

AMA Style

Laura De Lellis, Serena Veschi, Nicola Tinari, Zhirajr Mokini, Simone Carradori, Davide Brocco, Rosalba Florio, Antonino Grassadonia, Alessandro Cama. Drug Repurposing, an Attractive Strategy in Pancreatic Cancer Treatment: Preclinical and Clinical Updates. Cancers. 2021; 13 (16):3946.

Chicago/Turabian Style

Laura De Lellis; Serena Veschi; Nicola Tinari; Zhirajr Mokini; Simone Carradori; Davide Brocco; Rosalba Florio; Antonino Grassadonia; Alessandro Cama. 2021. "Drug Repurposing, an Attractive Strategy in Pancreatic Cancer Treatment: Preclinical and Clinical Updates." Cancers 13, no. 16: 3946.

Journal article
Published: 14 June 2021 in Pharmaceuticals
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This scientific research focused on the production of hydroethanolic extract of the plant species Lycopodium selago L. (L. selago) by the ultrasound-assisted extraction (USAE) and the identification of biocompounds with high antioxidant activity is of interest for possible phytotherapeutic treatment against Alzheimer’s disease (AD). The extract was phytochemically analyzed to investigate polyphenols, flavonoids, and identify the sesquiterpenoid alkaloid huperzine A (HupA), which is known in the literature for its great relevance in AD. Evaluation and comparison of the antioxidant activity of the extract were performed by four complementary spectrophotometric methods (DPPH, FRAP, ABTS, ORAC). In vitro tests of the extract showed an excellent reciprocal link between the concentration of polyphenols and the measurement of the antioxidant activity of the extract with the sesquiterpenoid HupA. To confirm the antioxidant activity, L. selago hydroethanolic extract was administered in vivo to zebrafish (Danio rerio) with a pattern of scopolamine-induced cognitive impairment. Moreover, this study explored a possible correlation between the expression of oxidative stress markers in the brain tissue with the behavior of the scopolamine zebrafish model. In vivo tests showed that this fern could be used as a nutritional supply and as a phytotherapeutic method to prevent or treat various neurodegenerative diseases that call for high-nutritive-value medications.

ACS Style

Mihai-Vlad Valu; Catalin Ducu; Sorin Moga; Denis Negrea; Lucian Hritcu; Razvan Boiangiu; Emanuel Vamanu; Tudor Balseanu; Simone Carradori; Liliana Soare. Effects of the Hydroethanolic Extract of Lycopodium selago L. on Scopolamine-Induced Memory Deficits in Zebrafish. Pharmaceuticals 2021, 14, 568 .

AMA Style

Mihai-Vlad Valu, Catalin Ducu, Sorin Moga, Denis Negrea, Lucian Hritcu, Razvan Boiangiu, Emanuel Vamanu, Tudor Balseanu, Simone Carradori, Liliana Soare. Effects of the Hydroethanolic Extract of Lycopodium selago L. on Scopolamine-Induced Memory Deficits in Zebrafish. Pharmaceuticals. 2021; 14 (6):568.

Chicago/Turabian Style

Mihai-Vlad Valu; Catalin Ducu; Sorin Moga; Denis Negrea; Lucian Hritcu; Razvan Boiangiu; Emanuel Vamanu; Tudor Balseanu; Simone Carradori; Liliana Soare. 2021. "Effects of the Hydroethanolic Extract of Lycopodium selago L. on Scopolamine-Induced Memory Deficits in Zebrafish." Pharmaceuticals 14, no. 6: 568.

Journal article
Published: 12 June 2021 in Journal of Fungi
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Hericium erinaceus (H. erinaceus) is a rare and appreciated fungal species belonging to the division Basidiomycota used for centuries in traditional Chinese medicine for its medicinal value. This species of mushrooms brings the most diverse benefits for the human body, and can have beneficial effects for treating Alzheimer’s disease (AD). This study investigated whether ethanolic extract from the fungal biomass of H. erinaceus enhances cognitive function via the action on cholinergic neurons using the scopolamine (SCOP)-induced zebrafish (Danio rerio) model of memory impairment. The ethanolic extract from the fungal biomass of H. erinaceus was previously obtained using an ultrasonic extraction method (UE). The administration of H. erinaceus extract to zebrafish, with a pattern of AD induced by scopolamine, showed an improvement in memory evaluated by behavioral and biochemical tests on brain tissue. These results suggest that H. erinaceus has preventive and therapeutic potentials in managing memory deficits and brain oxidative stress in zebrafish with AD.

ACS Style

Mihai-Vlad Valu; Liliana Soare; Catalin Ducu; Sorin Moga; Denis Negrea; Emanuel Vamanu; Tudor-Adrian Balseanu; Simone Carradori; Lucian Hritcu; Razvan Boiangiu. Hericium erinaceus (Bull.) Pers. Ethanolic Extract with Antioxidant Properties on Scopolamine-Induced Memory Deficits in a Zebrafish Model of Cognitive Impairment. Journal of Fungi 2021, 7, 477 .

AMA Style

Mihai-Vlad Valu, Liliana Soare, Catalin Ducu, Sorin Moga, Denis Negrea, Emanuel Vamanu, Tudor-Adrian Balseanu, Simone Carradori, Lucian Hritcu, Razvan Boiangiu. Hericium erinaceus (Bull.) Pers. Ethanolic Extract with Antioxidant Properties on Scopolamine-Induced Memory Deficits in a Zebrafish Model of Cognitive Impairment. Journal of Fungi. 2021; 7 (6):477.

Chicago/Turabian Style

Mihai-Vlad Valu; Liliana Soare; Catalin Ducu; Sorin Moga; Denis Negrea; Emanuel Vamanu; Tudor-Adrian Balseanu; Simone Carradori; Lucian Hritcu; Razvan Boiangiu. 2021. "Hericium erinaceus (Bull.) Pers. Ethanolic Extract with Antioxidant Properties on Scopolamine-Induced Memory Deficits in a Zebrafish Model of Cognitive Impairment." Journal of Fungi 7, no. 6: 477.

Journal article
Published: 04 June 2021 in Sustainability
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The collection of the organic fraction in Italy recorded significant growth in the decade 2010–2019 (+74%) due to both the increase in the spread of separate waste collection as well as the increase in the biological treatment plants of municipal waste. However, within the organic fraction there remains a share of non-compostable material (NCM) (plastic, glass, aluminum, etc.), equal to ~5% of the total, which affects the efficiency of composting practices as well as decreasing both the yield and the quality of the final compost, causing a portion of organic material to be subtracted from composting and ending up in landfills. Therefore, the purpose of this work is to evaluate how the sustainability of the organic fraction collection and the amount of compost obtained in the composting plants could improve, following the use of biodegradable and compostable bioplastic bags (shoppers), in replacement for conventional plastic ones. The Life Cycle Assessment (LCA) and Carbon Footprint (CF) methodology was used for the assessment, comparing two different scenarios based on data relating to the collection of the organic fraction in Italy in 2019. Scenario 1 relates to the composting of organic material that also contains plastics, bioplastics, and non-compostable materials, while in scenario 2 the share of plastic material in the first scenario has been entirely replaced by bioplastics. The results show that scenario 2 assumes the lowest values for 15 impact categories out of the 18 analyzed, and, among these, in three categories it assumes negative values: ozone formation-terrestrial ecosystems (−1.64 × 10−2 kg NOX eq), ozone formation-human health (−8.50 × 10−3 kg NOX eq), and fossil resource scarcity (−4.91 × 102 kg oil eq). Furthermore, scenario 2 has a negative carbon footprint (−3.80 kg CO2 eq) compared to scenario 1 (79.71 kg CO2 eq), and in general it is the most sustainable scenario as a direct consequence of the greater amount of compost obtained (307.4 kg vs. 269.2 kg).

ACS Style

Giuliana Vinci; Roberto Ruggieri; Andrea Billi; Carmine Pagnozzi; Maria Di Loreto; Marco Ruggeri. Sustainable Management of Organic Waste and Recycling for Bioplastics: A LCA Approach for the Italian Case Study. Sustainability 2021, 13, 6385 .

AMA Style

Giuliana Vinci, Roberto Ruggieri, Andrea Billi, Carmine Pagnozzi, Maria Di Loreto, Marco Ruggeri. Sustainable Management of Organic Waste and Recycling for Bioplastics: A LCA Approach for the Italian Case Study. Sustainability. 2021; 13 (11):6385.

Chicago/Turabian Style

Giuliana Vinci; Roberto Ruggieri; Andrea Billi; Carmine Pagnozzi; Maria Di Loreto; Marco Ruggeri. 2021. "Sustainable Management of Organic Waste and Recycling for Bioplastics: A LCA Approach for the Italian Case Study." Sustainability 13, no. 11: 6385.

Review
Published: 01 June 2021 in Expert Opinion on Drug Discovery
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Introduction: Among protein and fibers in the extracellular matrix (ECM), collagen is the most copious and widely employed in cosmetic, food, pharmaceutical, and biomedical industries due to its extensive biocompatible and versatile properties. In the last years, the knowledge about functions of collagens increased and expanded dramatically. Once considered only crucial for the ECM scaffolding and mechanotransduction, additional functional roles have now been ascribed to the collagen superfamily which are defined by other recently discovered domains, supramolecular assembly and receptors. Areas covered: Given the importance of each step in the collagen biosynthesis, folding and signaling, medicinal chemists have explored small molecules, peptides, and monoclonal antibodies to modulate enzymes, receptors and interactions with the physiological ligands of collagen. These compounds were also explored toward diseases and pathological conditions. The authors discuss this providing their expert perspectives on the subject area. Expert opinion: Understanding collagen protein properties and its interactome is beneficial for therapeutic drug design. Nevertheless, compounds targeting collagen-based interactome suffered from the presence of different isoforms for each target and the lack of specific 3D crystal structures able to guide properly drug design.

ACS Style

Marialucia Gallorini; Simone Carradori. Understanding collagen interactions and their targeted regulation by novel drugs. Expert Opinion on Drug Discovery 2021, 1 -22.

AMA Style

Marialucia Gallorini, Simone Carradori. Understanding collagen interactions and their targeted regulation by novel drugs. Expert Opinion on Drug Discovery. 2021; ():1-22.

Chicago/Turabian Style

Marialucia Gallorini; Simone Carradori. 2021. "Understanding collagen interactions and their targeted regulation by novel drugs." Expert Opinion on Drug Discovery , no. : 1-22.

Journal article
Published: 31 May 2021 in Current Medicinal Chemistry
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COVID-19, a type of infection that emerged in Wuhan, has become a pandemic affecting people worldwide and is rapidly spreading and evolving. Day by day, the confirmed cases and deaths are increasing many folds. SARS-CoV-2 is a novel virus; therefore, limited data are available to curb the disease. Epidemiological approaches, such as isolation, quarantine, social distancing, lockdown, and curfew, are being employed to halt the spread of the disease. Individual and joint efforts all over the world are producing a wealth of data and information which are expected to produce therapeutic strategies against COVID-19. Current research focuses on the utilization of antiviral drugs, repurposing strategies, vaccine development, as well as basic to advanced research about the organism and the infection. The review focuses on its life cycle, targets, and possible therapeutic strategies, which can lead to further research and development of COVID-19 therapy.

ACS Style

Rajesh Kumar; Seetha Harilal; Abdullah G. Al-Sehemi; Githa Elizabeth Mathew; Simone Carradori; Bijo Mathew. The Chronicle of COVID-19 and Possible Strategies to Curb the Pandemic. Current Medicinal Chemistry 2021, 28, 2852 -2886.

AMA Style

Rajesh Kumar, Seetha Harilal, Abdullah G. Al-Sehemi, Githa Elizabeth Mathew, Simone Carradori, Bijo Mathew. The Chronicle of COVID-19 and Possible Strategies to Curb the Pandemic. Current Medicinal Chemistry. 2021; 28 (15):2852-2886.

Chicago/Turabian Style

Rajesh Kumar; Seetha Harilal; Abdullah G. Al-Sehemi; Githa Elizabeth Mathew; Simone Carradori; Bijo Mathew. 2021. "The Chronicle of COVID-19 and Possible Strategies to Curb the Pandemic." Current Medicinal Chemistry 28, no. 15: 2852-2886.

Journal article
Published: 28 May 2021 in Current Medicinal Chemistry
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A few decades ago, the incidence of colorectal cancer (CRC) was low and is now the fourth in the list of deadly cancers producing nearly a million deaths annually. A population that is aging along with risk factors such as smoking, obesity, sedentary lifestyle with little or no physical activity, and non-healthy food habits of developed countries can increase the risk of colorectal cancer. The balance in gut microbiota and the metabolites produced during bacterial fermentation within the host plays a significant role in regulating intestinal diseases as well as colorectal cancer development. Recent progress in the understanding of illness resulted in multiple treatment options such as surgery, radiation, and chemotherapy, including targeted therapy and multitherapies. The treatment plan for CRC depends on the location, stage and grade of cancer as well as genomic biomarker tests. Despite all the advancements made in the genetic and molecular aspects of the disease, the knowledge seems inadequate as the drug action as well as the wide variation in drug response did not appear strongly correlated with the individual molecular and genetic characteristics, which suggests the requirement of comprehensive molecular understanding of this complex heterogeneous disease. Furthermore, multitherapies or a broad spectrum approach, which is an amalgamation of the various promising as well as effective therapeutic strategies that can tackle heterogeneity and act on several targets of the disease, need to be validated in clinical studies. The latest treatment options have significantly increased the survival of up to three years in the case of advanced disease. The fact that colorectal cancer is developed from a polypoid precursor, as well as the symptoms of the disease that occur at an advanced stage, underlines how screening programs can help early detection and decrease mortality as well as morbidity from CRC.

ACS Style

Rajesh Kumar; Seetha Harilal; Simone Carradori; Bijo Mathew. A Comprehensive Overview of Colon Cancer- A Grim Reaper of the 21st Century. Current Medicinal Chemistry 2021, 28, 2657 -2696.

AMA Style

Rajesh Kumar, Seetha Harilal, Simone Carradori, Bijo Mathew. A Comprehensive Overview of Colon Cancer- A Grim Reaper of the 21st Century. Current Medicinal Chemistry. 2021; 28 (14):2657-2696.

Chicago/Turabian Style

Rajesh Kumar; Seetha Harilal; Simone Carradori; Bijo Mathew. 2021. "A Comprehensive Overview of Colon Cancer- A Grim Reaper of the 21st Century." Current Medicinal Chemistry 28, no. 14: 2657-2696.

Journal article
Published: 14 May 2021 in Molecules
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The chemical profile of the female inflorescence extracts from seven Cannabis sativa L. dioecious cultivars (Carmagnola, Fibranova, Eletta Campana, Antal, Tiborszallasi, Kompolti, and Tisza) was monitored at three harvesting stages (4, 14, and 30 September), reaching from the beginning of flowering to end of flowering/beginning of seed formation, using untargeted nuclear magnetic resonance (NMR) and targeted (ultra-high-performance liquid chromatography (UHPLC) and spectrophotometry) analyses. The tetrahydrocannabinol content was always below the legal limits (<0.6%) in all the analyzed samples. The NMR metabolite profile (sugars, organic acids, amino acids, and minor compounds) subjected to principal components analysis (PCA) showed a strong variability according to the harvesting stages: samples harvested in stage I were characterized by a high content of sucrose and myo-inositol, whereas the ones harvested in stage II showed high levels of succinic acid, alanine, valine, isoleucine, phenylalanine, and threonine. Samples harvested in stage III were characterized by high levels of glucose, fructose, choline, trigonelline, malic acid, formic acid, and some amino acids. The ratio between chlorophylls and carotenoids content indicated that all plants grew up exposed to the sun, the Eletta Campana cultivar having the highest pigment amount. Tiborszallasi cultivar showed the highest polyphenol content. The highest antioxidant activity was generally observed in stage II. All these results suggested that the Cannabis sativa L. inflorescences of each analyzed dioecious hemp cultivar presented a peculiar chemical profile affected by the harvesting stage. This information could be useful for producers and industries to harvest inflorescences in the appropriate stage to obtain samples with a peculiar chemical profile suitable for proper applications.

ACS Style

Mattia Spano; Giacomo Di Matteo; Cinzia Ingallina; Bruno Botta; Deborah Quaglio; Francesca Ghirga; Silvia Balducci; Silvia Cammarone; Enio Campiglia; Anna Giusti; Giuliana Vinci; Mattia Rapa; Salvatore Ciano; Luisa Mannina; Anatoly Sobolev. A Multimethodological Characterization of Cannabis sativa L. Inflorescences from Seven Dioecious Cultivars Grown in Italy: The Effect of Different Harvesting Stages. Molecules 2021, 26, 2912 .

AMA Style

Mattia Spano, Giacomo Di Matteo, Cinzia Ingallina, Bruno Botta, Deborah Quaglio, Francesca Ghirga, Silvia Balducci, Silvia Cammarone, Enio Campiglia, Anna Giusti, Giuliana Vinci, Mattia Rapa, Salvatore Ciano, Luisa Mannina, Anatoly Sobolev. A Multimethodological Characterization of Cannabis sativa L. Inflorescences from Seven Dioecious Cultivars Grown in Italy: The Effect of Different Harvesting Stages. Molecules. 2021; 26 (10):2912.

Chicago/Turabian Style

Mattia Spano; Giacomo Di Matteo; Cinzia Ingallina; Bruno Botta; Deborah Quaglio; Francesca Ghirga; Silvia Balducci; Silvia Cammarone; Enio Campiglia; Anna Giusti; Giuliana Vinci; Mattia Rapa; Salvatore Ciano; Luisa Mannina; Anatoly Sobolev. 2021. "A Multimethodological Characterization of Cannabis sativa L. Inflorescences from Seven Dioecious Cultivars Grown in Italy: The Effect of Different Harvesting Stages." Molecules 26, no. 10: 2912.

Editorial
Published: 07 May 2021 in Molecules
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The aim of the present special issue, proposed by two Co-Guest Editors with expertise in Clinical Microbiology and Medicinal Chemistry, is to collect and disseminate some of the most significant and innovative contributions focused on biofilm removal strategies, based on the use of natural or synthetic compounds/molecules/peptides or nanoparticles as well as biofilm formation inhibition aimed at both the control and monitoring of biofilm infections in medicine, food, industry, and natural environments

ACS Style

Rossella Grande; Simone Carradori. Novel Biologically Active Molecules, Biomaterials, and Nanoparticles for Microbial Biofilm Control in Human Medicine. Molecules 2021, 26, 2749 .

AMA Style

Rossella Grande, Simone Carradori. Novel Biologically Active Molecules, Biomaterials, and Nanoparticles for Microbial Biofilm Control in Human Medicine. Molecules. 2021; 26 (9):2749.

Chicago/Turabian Style

Rossella Grande; Simone Carradori. 2021. "Novel Biologically Active Molecules, Biomaterials, and Nanoparticles for Microbial Biofilm Control in Human Medicine." Molecules 26, no. 9: 2749.

Journal article
Published: 17 April 2021 in Pharmaceuticals
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Repurposing of approved non-antitumor drugs represents a promising and affordable strategy that may help to increase the repertoire of effective anticancer drugs. Benzimidazole-based anthelmintics are antiparasitic drugs commonly employed both in human and veterinary medicine. Benzimidazole compounds are being considered for drug repurposing due to antitumor activities displayed by some members of the family. In this study, we explored the effects of a large series of benzimidazole-based anthelmintics (and some enantiomerically pure forms of those containing a stereogenic center) on the viability of different tumor cell lines derived from paraganglioma, pancreatic and colorectal cancer. Flubendazole, parbendazole, oxibendazole, mebendazole, albendazole and fenbendazole showed the most consistent antiproliferative effects, displaying IC50 values in the low micromolar range, or even in the nanomolar range. In silico evaluation of their physicochemical, pharmacokinetics and medicinal chemistry properties also provided useful information related to the chemical structures and potential of these compounds. Furthermore, in view of the potential repurposing of these drugs in cancer therapy and considering that pharmaceutically active compounds may have different mechanisms of action, we performed an in silico target prediction to assess the polypharmacology of these benzimidazoles, which highlighted previously unknown cancer-relevant molecular targets.

ACS Style

Rosalba Florio; Simone Carradori; Serena Veschi; Davide Brocco; Teresa Di Genni; Roberto Cirilli; Adriano Casulli; Alessandro Cama; Laura De Lellis. Screening of Benzimidazole-Based Anthelmintics and Their Enantiomers as Repurposed Drug Candidates in Cancer Therapy. Pharmaceuticals 2021, 14, 372 .

AMA Style

Rosalba Florio, Simone Carradori, Serena Veschi, Davide Brocco, Teresa Di Genni, Roberto Cirilli, Adriano Casulli, Alessandro Cama, Laura De Lellis. Screening of Benzimidazole-Based Anthelmintics and Their Enantiomers as Repurposed Drug Candidates in Cancer Therapy. Pharmaceuticals. 2021; 14 (4):372.

Chicago/Turabian Style

Rosalba Florio; Simone Carradori; Serena Veschi; Davide Brocco; Teresa Di Genni; Roberto Cirilli; Adriano Casulli; Alessandro Cama; Laura De Lellis. 2021. "Screening of Benzimidazole-Based Anthelmintics and Their Enantiomers as Repurposed Drug Candidates in Cancer Therapy." Pharmaceuticals 14, no. 4: 372.

Journal article
Published: 24 March 2021 in Molecules
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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.

ACS Style

Francesca Sisto; Simone Carradori; Paolo Guglielmi; Mattia Spano; Daniela Secci; Arianna Granese; Anatoly Sobolev; Rossella Grande; Cristina Campestre; Maria Di Marcantonio; Gabriella Mincione. Synthesis and Evaluation of Thymol-Based Synthetic Derivatives as Dual-Action Inhibitors against Different Strains of H. pylori and AGS Cell Line. Molecules 2021, 26, 1829 .

AMA Style

Francesca Sisto, Simone Carradori, Paolo Guglielmi, Mattia Spano, Daniela Secci, Arianna Granese, Anatoly Sobolev, Rossella Grande, Cristina Campestre, Maria Di Marcantonio, Gabriella Mincione. Synthesis and Evaluation of Thymol-Based Synthetic Derivatives as Dual-Action Inhibitors against Different Strains of H. pylori and AGS Cell Line. Molecules. 2021; 26 (7):1829.

Chicago/Turabian Style

Francesca Sisto; Simone Carradori; Paolo Guglielmi; Mattia Spano; Daniela Secci; Arianna Granese; Anatoly Sobolev; Rossella Grande; Cristina Campestre; Maria Di Marcantonio; Gabriella Mincione. 2021. "Synthesis and Evaluation of Thymol-Based Synthetic Derivatives as Dual-Action Inhibitors against Different Strains of H. pylori and AGS Cell Line." Molecules 26, no. 7: 1829.

Journal article
Published: 03 March 2021 in Biomolecules
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In this paper, the first phytochemical analysis of the ethanolic extract of Daphne sericea Vahl flowering aerial parts collected in Italy and its biological activities were reported. Eleven compounds were identified i.e., α-linolenic acid (1), tri-linoleoyl-sn-glycerol (2), pheophorbide a ethyl ester (3), pilloin (4), sinensetin (5), yuanhuanin (6), rutamontine (7), syringin (8), p-coumaric acid (9), p-anisic acid (10) and caffeic acid (11). To the best of our knowledge, compounds (1-4, 7-8 and 10) were isolated from D. sericea for the first time during this work, whereas sinensetin (5) represents a newly identified component of the entire Thymelaeaceae family. The extract was found to possess radical scavenging against both DPPH• and 2,2′-azino-bis(3-thylbenzothiazoline-6-sulfonic acid (ABTS•+) radicals, with at least a 40-fold higher potency against the latter. Moreover, chelating abilities against both ferrous and ferric ions have been highlighted, thus suggesting a possible indirect antioxidant power of the extract. Although the precise bioactive compounds remain to be discovered, the polyphenolic constituents, including phenolic acids, tannins and flavonoids, seem to contribute to the antioxidant power of the phytocomplex. In addition, the extract produced cytotoxic effects in MDA-MB-231 and U87-MG cancer cell lines, especially at the concentration of 625 μg/mL and after 48–72 h. Further studies are required to clarify the contribution of the identified compounds in the bioactivities of the extract and to support possible future applications.

ACS Style

Claudio Frezza; Alessandro Venditti; Daniela De Vita; Fabio Sciubba; Pierpaolo Tomai; Marco Franceschin; Mirella Di Cecco; Giampiero Ciaschetti; Antonella Di Sotto; Annarita Stringaro; Marisa Colone; Alessandra Gentili; Mauro Serafini; Armandodoriano Bianco. Phytochemical Analysis and Biological Activities of the Ethanolic Extract of Daphne sericea Vahl Flowering Aerial Parts Collected in Central Italy. Biomolecules 2021, 11, 379 .

AMA Style

Claudio Frezza, Alessandro Venditti, Daniela De Vita, Fabio Sciubba, Pierpaolo Tomai, Marco Franceschin, Mirella Di Cecco, Giampiero Ciaschetti, Antonella Di Sotto, Annarita Stringaro, Marisa Colone, Alessandra Gentili, Mauro Serafini, Armandodoriano Bianco. Phytochemical Analysis and Biological Activities of the Ethanolic Extract of Daphne sericea Vahl Flowering Aerial Parts Collected in Central Italy. Biomolecules. 2021; 11 (3):379.

Chicago/Turabian Style

Claudio Frezza; Alessandro Venditti; Daniela De Vita; Fabio Sciubba; Pierpaolo Tomai; Marco Franceschin; Mirella Di Cecco; Giampiero Ciaschetti; Antonella Di Sotto; Annarita Stringaro; Marisa Colone; Alessandra Gentili; Mauro Serafini; Armandodoriano Bianco. 2021. "Phytochemical Analysis and Biological Activities of the Ethanolic Extract of Daphne sericea Vahl Flowering Aerial Parts Collected in Central Italy." Biomolecules 11, no. 3: 379.

Journal article
Published: 25 February 2021 in Pharmaceuticals
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Novel twenty-three 3(2H)-pyridazinone derivatives were designed and synthesized based on the chemical requirements related to the anti-proliferative effects previously demonstrated within this scaffold. The introduction of a piperazinyl linker between the pyridazinone nucleus and the additional (un)substituted phenyl group led to some compounds endowed with a limited cytotoxicity against human gingival fibroblasts (HGFs) and good anti-proliferative effects against gastric adenocarcinoma cells (AGS) as evaluated by MTT and LDH assays, using doxorubicin as a positive control. Successive analyses revealed that the two most promising representative compounds (12 and 22) could exert their effects by inducing oxidative stress as demonstrated by the hydrogen peroxide release and the morphological changes (cell blebbing) revealed by light microscopy analysis after the haematoxylin-eosin staining. Moreover, to further assess the apoptotic process induced by compounds 12 and 22, Bax expression was measured by flow cytometry. These findings enlarged our knowledge of the structural requirements in this scaffold to display valuable biological effects against cancerous cell lines.

ACS Style

Mehmet Alagöz; Zeynep Özdemir; Mehtap Uysal; Simone Carradori; Marialucia Gallorini; Alessia Ricci; Susi Zara; Bijo Mathew. Synthesis, Cytotoxicity and Anti-Proliferative Activity against AGS Cells of New 3(2H)-Pyridazinone Derivatives Endowed with a Piperazinyl Linker. Pharmaceuticals 2021, 14, 183 .

AMA Style

Mehmet Alagöz, Zeynep Özdemir, Mehtap Uysal, Simone Carradori, Marialucia Gallorini, Alessia Ricci, Susi Zara, Bijo Mathew. Synthesis, Cytotoxicity and Anti-Proliferative Activity against AGS Cells of New 3(2H)-Pyridazinone Derivatives Endowed with a Piperazinyl Linker. Pharmaceuticals. 2021; 14 (3):183.

Chicago/Turabian Style

Mehmet Alagöz; Zeynep Özdemir; Mehtap Uysal; Simone Carradori; Marialucia Gallorini; Alessia Ricci; Susi Zara; Bijo Mathew. 2021. "Synthesis, Cytotoxicity and Anti-Proliferative Activity against AGS Cells of New 3(2H)-Pyridazinone Derivatives Endowed with a Piperazinyl Linker." Pharmaceuticals 14, no. 3: 183.

Journal article
Published: 12 February 2021 in Biomolecules
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Industrial hemp is a multiuse crop that has been widely cultivated to produce fibers and nutrients. The capability of the essential oil (EO) from inflorescences as antimicrobial agent has been reported. However, literature data are still lacking about the hemp EO antiprotozoal efficacy in vivo. The present study aims to unravel this concern through the evaluation of the efficacy of hemp EOs (2.5 mL/kg, intraperitoneally) of three different cultivars, namely Futura 75, Carmagnola selezionata and Eletta campana, in mice intraperitoneally infected with Leishmania tropica. A detailed description of EO composition and targets-components analysis is reported. Myrcene, α-pinene and E-caryophyllene were the main components of the EOs, as indicated by the gas-chromatographic analysis. However, a prominent position in the scenario of the theoretical interactions underlying the bio-pharmacological activity was also occupied by selina-3,7(11)-diene, which displayed affinities in the micromolar range (5.4–28.9) towards proliferator-activated receptor α, cannabinoid CB2 receptor and acetylcholinesterase. The content of this compound was higher in Futura 75 and Eletta campana, in accordance with their higher scavenging/reducing properties and efficacy against the tissue wound, induced by L. tropica. Overall, the present study recommends hemp female inflorescences, as sources of biomolecules with potential pharmacological applications, especially towards infective diseases.

ACS Style

Luigi Menghini; Claudio Ferrante; Simone Carradori; Marianna D’Antonio; Giustino Orlando; Francesco Cairone; Stefania Cesa; Antonello Filippi; Caterina Fraschetti; Gokhan Zengin; Gunes Ak; Massimo Tacchini; Kashif Iqbal. Chemical and Bioinformatics Analyses of the Anti-Leishmanial and Anti-Oxidant Activities of Hemp Essential Oil. Biomolecules 2021, 11, 272 .

AMA Style

Luigi Menghini, Claudio Ferrante, Simone Carradori, Marianna D’Antonio, Giustino Orlando, Francesco Cairone, Stefania Cesa, Antonello Filippi, Caterina Fraschetti, Gokhan Zengin, Gunes Ak, Massimo Tacchini, Kashif Iqbal. Chemical and Bioinformatics Analyses of the Anti-Leishmanial and Anti-Oxidant Activities of Hemp Essential Oil. Biomolecules. 2021; 11 (2):272.

Chicago/Turabian Style

Luigi Menghini; Claudio Ferrante; Simone Carradori; Marianna D’Antonio; Giustino Orlando; Francesco Cairone; Stefania Cesa; Antonello Filippi; Caterina Fraschetti; Gokhan Zengin; Gunes Ak; Massimo Tacchini; Kashif Iqbal. 2021. "Chemical and Bioinformatics Analyses of the Anti-Leishmanial and Anti-Oxidant Activities of Hemp Essential Oil." Biomolecules 11, no. 2: 272.

Journal article
Published: 01 February 2021 in Biomedicines
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Sustained oxidative stress and inflammation have been reported as the major factors responsible for the failure of tendon healing during rotator cuff tears (RCTs) and rotator cuff disease (RCD). Although, their therapeutic management remains still challenging. Carbonic anhydrases (CAs) are involved in many pathological conditions, and the overexpression of both CA9 and 12 in inflamed joints has been recently reported. Consequently, a selective CA9/12 inhibition could be a feasible strategy for improving tendon recovery after injury. In addition, since carbon monoxide (CO) has been proven to have an important role in modulating inflammation, CO releasing molecules (CORMs) can be also potentially suitable compounds. The present study aims at evaluating five newly synthesized dual-mode acting CA inhibitors (CAIs)-CORMs compounds, belonging to two chemical scaffolds, on tendon-derived human primary cells under H2O2 stimulation in comparison with Meloxicam. Our results show that compounds 2 and 7 are the most promising of the series in counteracting oxidative stress-induced cytotoxicity and display a better profile in terms of enhanced viability, decreased LDH release, and augmented tenocyte proliferation compared to Meloxicam. Moreover, compound 7, as a potent superoxide scavenger, exerts its action inhibiting NF-ĸB translocation and downregulating iNOS, whereas compound 2 is more effective in increasing collagen I deposition. Taken together, our data highlight a potential role of CA in RCTs and RCD and the prospective effectiveness of compounds acting as CAI-CORM during inflammation.

ACS Style

Marialucia Gallorini; Anna C. Berardi; Alessia Ricci; Cristina Antonetti Lamorgese Passeri; Susi Zara; Francesco Oliva; Amelia Cataldi; Fabrizio Carta; Simone Carradori. Dual Acting Carbon Monoxide Releasing Molecules and Carbonic Anhydrase Inhibitors Differentially Modulate Inflammation in Human Tenocytes. Biomedicines 2021, 9, 141 .

AMA Style

Marialucia Gallorini, Anna C. Berardi, Alessia Ricci, Cristina Antonetti Lamorgese Passeri, Susi Zara, Francesco Oliva, Amelia Cataldi, Fabrizio Carta, Simone Carradori. Dual Acting Carbon Monoxide Releasing Molecules and Carbonic Anhydrase Inhibitors Differentially Modulate Inflammation in Human Tenocytes. Biomedicines. 2021; 9 (2):141.

Chicago/Turabian Style

Marialucia Gallorini; Anna C. Berardi; Alessia Ricci; Cristina Antonetti Lamorgese Passeri; Susi Zara; Francesco Oliva; Amelia Cataldi; Fabrizio Carta; Simone Carradori. 2021. "Dual Acting Carbon Monoxide Releasing Molecules and Carbonic Anhydrase Inhibitors Differentially Modulate Inflammation in Human Tenocytes." Biomedicines 9, no. 2: 141.

Review
Published: 08 January 2021 in Energies
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According to the United Nations (UN), although cities occupy only 3% of Earth’s surface, they host more than half of the global population, are responsible for 70% of energy consumption, and 75% of carbon emissions. All this is a consequence of the massive urbanization verified since the 1950s and which is expected to continue in the coming decades. A crucial issue will therefore concern the management of existing cities and the planning of future ones, and this was also emphasized by the UN Sustainable Development Goals (SDGs), especially in Goal 11 (Sustainable Cities and communities). Smart Cities are often seen as ideal urban environments in which the different dimensions of a city (economy, education, energy, environment, etc.) are managed successfully and proactively. So, one of the most important challenges cities will have to face, is to guide citizens towards a form of “clean” energy consumption, and the dimension on which decision-makers will be able to work is the decarbonization of transport. To achieve this, electric mobility could help reduce polluting emissions on the road. Within this research, the strategies that six Smart Cities (London, Hamburg, Oslo, Milan, Florence, and Bologna) have implemented to encourage the transition to this form of mobility have been studied. Through a systematic review of the literature (Scopus, Google Scholar, and Web of Science) and through the study of the main political/energy documents of the cities, their policies on electric mobility have been evaluated. Then, for each city, SDG 11.6.2 was analyzed to assess the air quality in the last four years (2016–2019) and, therefore, the effectiveness of the policies. The analysis showed, in general, that the policies have worked, inducing reductions in the pollutants of PM2.5, PM10, NO2. In particular, the cities showed the most significant reduction in pollutant (above 20%) were Hamburg (−28% PM2.5 and −2%6 NO2), Milan (−25% PM2.5 and −52% NO2), and London (−26% NO2).

ACS Style

Roberto Ruggieri; Marco Ruggeri; Giuliana Vinci; Stefano Poponi. Electric Mobility in a Smart City: European Overview. Energies 2021, 14, 315 .

AMA Style

Roberto Ruggieri, Marco Ruggeri, Giuliana Vinci, Stefano Poponi. Electric Mobility in a Smart City: European Overview. Energies. 2021; 14 (2):315.

Chicago/Turabian Style

Roberto Ruggieri; Marco Ruggeri; Giuliana Vinci; Stefano Poponi. 2021. "Electric Mobility in a Smart City: European Overview." Energies 14, no. 2: 315.

Journal article
Published: 05 January 2021 in Antioxidants
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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.

ACS Style

Emanuela Berrino; Simone Carradori; Andrea Angeli; Fabrizio Carta; Claudiu Supuran; Paolo Guglielmi; Cecilia Coletti; Roberto Paciotti; Helmut Schweikl; Francesca Maestrelli; Elisabetta Cerbai; Marialucia Gallorini. Dual Carbonic Anhydrase IX/XII Inhibitors and Carbon Monoxide Releasing Molecules Modulate LPS-Mediated Inflammation in Mouse Macrophages. Antioxidants 2021, 10, 56 .

AMA Style

Emanuela Berrino, Simone Carradori, Andrea Angeli, Fabrizio Carta, Claudiu Supuran, Paolo Guglielmi, Cecilia Coletti, Roberto Paciotti, Helmut Schweikl, Francesca Maestrelli, Elisabetta Cerbai, Marialucia Gallorini. Dual Carbonic Anhydrase IX/XII Inhibitors and Carbon Monoxide Releasing Molecules Modulate LPS-Mediated Inflammation in Mouse Macrophages. Antioxidants. 2021; 10 (1):56.

Chicago/Turabian Style

Emanuela Berrino; Simone Carradori; Andrea Angeli; Fabrizio Carta; Claudiu Supuran; Paolo Guglielmi; Cecilia Coletti; Roberto Paciotti; Helmut Schweikl; Francesca Maestrelli; Elisabetta Cerbai; Marialucia Gallorini. 2021. "Dual Carbonic Anhydrase IX/XII Inhibitors and Carbon Monoxide Releasing Molecules Modulate LPS-Mediated Inflammation in Mouse Macrophages." Antioxidants 10, no. 1: 56.

Journal article
Published: 01 January 2021 in Antioxidants
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Industrial hemp is a multiuse crop whose phytocomplex includes terpenophenolics and flavonoids. In the present study, the phenolic and terpenophenolic compounds were assayed in the water extract of the hemp variety Futura 75. Protective effects were also investigated in human fibroblast and keratinocytes and isolate mouse skin specimens, which were exposed to hydrogen peroxide and/or to the extract (1–500 µg/mL). The results of phytochemical analysis suggested the cannabidiol, cannabidiolic acid and rutin as the prominent phytocompounds. In the in vitro system represented by human keratinocytes and fibroblasts, the hemp extract was found to be able to protect cells from cytotoxicity and apoptosis induced by oxidative stress. Moreover, modulatory effects on IL-6, a key mediator in skin proliferation, were found. In isolated rat skin, the extract reduced hydrogen peroxide-induced l-dopa turnover, prostaglandin-E2 production and the ratio kynurenine/tryptpophan, thus corroborating anti-inflammatory/antioxidant effects. The in silico docking studies also highlighted the putative interactions between cannabidiol, cannabidiolic acid and rutin with tyrosinase and indoleamine-2,3-dioxygenase, involved in l-dopa turnover and tryptophan conversion in kynurenine, respectively. In conclusion, the present findings showed the efficacy of hemp water extract as a skin protective agent. This could be partly related to the extract content in cannabidiol, cannabidiolic acid and rutin.

ACS Style

Viviana Di Giacomo; Lucia Recinella; Annalisa Chiavaroli; Giustino Orlando; Amelia Cataldi; Monica Rapino; Valentina Di Valerio; Matteo Politi; Marco Daniel Antolini; Alessandra Acquaviva; Francesco Bacchin; Massimo Di Mascio; Sheila Leone; Luigi Brunetti; Luigi Menghini; Simone Carradori; Gokhan Zengin; Gunes Ak; Claudio Ferrante. Metabolomic Profile and Antioxidant/Anti-Inflammatory Effects of Industrial Hemp Water Extract in Fibroblasts, Keratinocytes and Isolated Mouse Skin Specimens. Antioxidants 2021, 10, 44 .

AMA Style

Viviana Di Giacomo, Lucia Recinella, Annalisa Chiavaroli, Giustino Orlando, Amelia Cataldi, Monica Rapino, Valentina Di Valerio, Matteo Politi, Marco Daniel Antolini, Alessandra Acquaviva, Francesco Bacchin, Massimo Di Mascio, Sheila Leone, Luigi Brunetti, Luigi Menghini, Simone Carradori, Gokhan Zengin, Gunes Ak, Claudio Ferrante. Metabolomic Profile and Antioxidant/Anti-Inflammatory Effects of Industrial Hemp Water Extract in Fibroblasts, Keratinocytes and Isolated Mouse Skin Specimens. Antioxidants. 2021; 10 (1):44.

Chicago/Turabian Style

Viviana Di Giacomo; Lucia Recinella; Annalisa Chiavaroli; Giustino Orlando; Amelia Cataldi; Monica Rapino; Valentina Di Valerio; Matteo Politi; Marco Daniel Antolini; Alessandra Acquaviva; Francesco Bacchin; Massimo Di Mascio; Sheila Leone; Luigi Brunetti; Luigi Menghini; Simone Carradori; Gokhan Zengin; Gunes Ak; Claudio Ferrante. 2021. "Metabolomic Profile and Antioxidant/Anti-Inflammatory Effects of Industrial Hemp Water Extract in Fibroblasts, Keratinocytes and Isolated Mouse Skin Specimens." Antioxidants 10, no. 1: 44.

Review article
Published: 01 January 2021 in Journal of Enzyme Inhibition and Medicinal Chemistry
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Inorganic anions inhibit the metalloenzyme carbonic anhydrase (CA, EC 4.2.1.1) generally by coordinating to the active site metal ion. Cyanate was reported as a non-coordinating CA inhibitor but those erroneous results were subsequently corrected by another group. We review the anion CA inhibitors (CAIs) in the more general context of drug design studies and the discovery of a large number of inhibitor classes and inhibition mechanisms, including zinc binders (sulphonamides and isosteres, dithiocabamates and isosteres, thiols, selenols, benzoxaboroles, ninhydrins, etc.); inhibitors anchoring to the zinc-coordinated water molecule (phenols, polyamines, sulfocoumarins, thioxocoumarins, catechols); CAIs occluding the entrance to the active site (coumarins and derivatives, lacosamide), as well as compounds that bind outside the active site. All these new chemotypes integrated with a general procedure for obtaining isoform-selective compounds (the tail approach) has resulted, through the guidance of rigorous X-ray crystallography experiments, in the development of highly selective CAIs for all human CA isoforms with many pharmacological applications.

ACS Style

Alessio Nocentini; Andrea Angeli; Fabrizio Carta; Jean-Yves Winum; Raivis Zalubovskis; Simone Carradori; Clemente Capasso; William A. Donald; Claudiu T. Supuran. Reconsidering anion inhibitors in the general context of drug design studies of modulators of activity of the classical enzyme carbonic anhydrase. Journal of Enzyme Inhibition and Medicinal Chemistry 2021, 36, 561 -580.

AMA Style

Alessio Nocentini, Andrea Angeli, Fabrizio Carta, Jean-Yves Winum, Raivis Zalubovskis, Simone Carradori, Clemente Capasso, William A. Donald, Claudiu T. Supuran. Reconsidering anion inhibitors in the general context of drug design studies of modulators of activity of the classical enzyme carbonic anhydrase. Journal of Enzyme Inhibition and Medicinal Chemistry. 2021; 36 (1):561-580.

Chicago/Turabian Style

Alessio Nocentini; Andrea Angeli; Fabrizio Carta; Jean-Yves Winum; Raivis Zalubovskis; Simone Carradori; Clemente Capasso; William A. Donald; Claudiu T. Supuran. 2021. "Reconsidering anion inhibitors in the general context of drug design studies of modulators of activity of the classical enzyme carbonic anhydrase." Journal of Enzyme Inhibition and Medicinal Chemistry 36, no. 1: 561-580.

Journal article
Published: 30 December 2020 in Current Medicinal Chemistry
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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.

ACS Style

Bijo Mathew; Simone Carradori; Paolo Guglielmi; Sahab Uddin; Hoon Kim. New Aspects of Monoamine Oxidase B Inhibitors: The Key Role of Halogens to Open the Golden Door. Current Medicinal Chemistry 2020, 28, 266 -283.

AMA Style

Bijo Mathew, Simone Carradori, Paolo Guglielmi, Sahab Uddin, Hoon Kim. New Aspects of Monoamine Oxidase B Inhibitors: The Key Role of Halogens to Open the Golden Door. Current Medicinal Chemistry. 2020; 28 (2):266-283.

Chicago/Turabian Style

Bijo Mathew; Simone Carradori; Paolo Guglielmi; Sahab Uddin; Hoon Kim. 2020. "New Aspects of Monoamine Oxidase B Inhibitors: The Key Role of Halogens to Open the Golden Door." Current Medicinal Chemistry 28, no. 2: 266-283.