Leveraging SARS-CoV-2 Main Protease (Mpro) for COVID-19 Mitigation with Selenium-Based Inhibitors.

The implementation of innovative approaches is crucial in an ongoing endeavor to mitigate the impact of COVID-19 pandemic. The present study examines the strategic application of the SARS-CoV-2 Main Protease (Mpro) as a prospective instrument in the repertoire to combat the virus. The cloning, expression, and purification of Mpro, which plays a critical role in the viral life cycle, through heterologous expression in Escherichia coli in a completely soluble form produced an active enzyme. The hydrolysis of a specific substrate peptide comprising a six-amino-acid sequence (TSAVLQ) linked to a p-nitroaniline (pNA) fragment together with the use of a fluorogenic substrate allowed us to determine effective inhibitors incorporating selenium moieties, such as benzoselenoates and carbamoselenoates. The new inhibitors revealed their potential to proficiently inhibit Mpro with IC50-s in the low micromolar range. Our study contributes to the development of a new class of protease inhibitors targeting Mpro, ultimately strengthening the antiviral arsenal against COVID-19 and possibly, related coronaviruses.

Nature as a source and inspiration for human monoamine oxidase B (hMAO-B) inhibition: A review of the recent advances in chemical modification of natural compounds.

INTRODUCTION: Over the past 5 years, we have witnessed intense research activity about the biological potential of natural products (NPs) as human monoamine oxidase B (hMAO-B) inhibitors. Despite the promising inhibitory activity, natural compounds often suffer from pharmacokinetic lissues, such as poor aqueous solubility, extensive metabolism, and low bioavailability. AREAS COVERED: This review provides an overview of the current landscape NPs as selective hMAO-B inhibitors and highlights their use as a starting scaffold to design (semi)synthetic derivatives to overcome the therapeutic (pharmacodynamic and pharmacokinetic) limitations of NPs and to obtain more robust structure-activity relationships (SARs) for each scaffold. EXPERT OPINION: All the natural scaffolds herein presented displayed a broad chemical diversity. The knowledge of their biological activity as inhibitors of hMAO-B enzyme allows the positive correlations associated with the consumption of specific food or the possible herb-drug interactions and suggests to the Medicinal Chemists how to address chemical functionalization to obtain more potent and selective compounds.

Evaluation of a dill (Anethum graveolens L.) gene bank germplasm collection using multivariate analysis of morphological traits, molecular genotyping and chemical composition to identify novel genotypes for plant breeding.

Dill (Anethum graveolens L.) is an aromatic herb widely used in the food industry, with several commercial cultivars available with different qualitative characteristics. Commercial cultivars are usually preferred over landraces due to their higher yield and also the lack of improved landraces than can be commercialized. In Greece, however, traditional dill landraces are cultivated by local communities. Many are conserved in the Greek Gene Bank and the aim here was to investigate and compare the morphological, genetic, and chemical biodiversity of twenty-two Greek landraces and nine modern/commercial cultivars. Multivariate analysis of the morphological descriptors, molecular markers, and essential oil and polyphenol composition revealed that the Greek landraces were clearly distinguished compared with modern cultivars at the level of phenological, molecular and chemical traits. Landraces were typically taller, with larger umbels, denser foliage, and larger leaves. Plant height, density of foliage, density of feathering as well as aroma characteristics were desirable traits observed for some landraces, such as T538/06 and GRC-1348/04, which were similar or superior to those of some commercial cultivars. Polymorphic loci for inter-simple sequence repeat (ISSR) and start codon targeted (SCoT) molecular markers were 76.47% and 72.41% for landraces, and 68.24% and 43.10% for the modern cultivars, respectively. Genetic divergence was shown, but not complete isolation, indicating that some gene flow may have occurred between landraces and cultivars. The major constituent in all dill leaf essential oils was α-phellandrene (54.42-70.25%). Landraces had a higher α-phellandrene and dill ether content than cultivars. Two dill landraces were rich in chlorogenic acid, the main polyphenolic compound determined. The study highlighted for the first-time Greek landraces with desirable characteristics regarding quality, yield, and harvest time suitable for breeding programs to develop new dill cultivars with superior features.

Selenium-analogs based on natural sources as cancer-associated carbonic anhydrase isoforms IX and XII inhibitors.

In the relentless search for new cancer treatments, organoselenium compounds, and carbonic anhydrase (CA) inhibitors have emerged as promising drug candidates. CA isoforms IX and XII are overexpressed in many types of cancer, and their inhibition is associated with potent antitumor/antimetastatic effects. Selenium-containing compounds, particularly selenols, have been shown to inhibit tumour-associated CA isoforms in the nanomolar range since the properties of the selenium atom favour binding to the active site of the enzyme. In this work, two series of selenoesters (1a-19a and 1b-19b), which gathered NSAIDs, carbo/heterocycles, and fragments from natural products, were evaluated against hCA I, II, IX, and XII. Indomethacin (17b) and flufenamic acid (19b) analogs exhibited selectivity for tumour-associated isoform IX in the low micromolar range. In summary, selenoesters that combine NSAIDs with fragments derived from natural sources have been developed as promising nonclassical inhibitors of the tumour-associated CA isoforms.

Vagar e ocupar: dez anos de narrativas no TOCCA - saberes e práticas transdisciplinares entre as artes e a saúde / Wander and occupy: 10 years of narratives in the TOCCA program - knowledge and practices in arts and health / Desocupar y ocupar: 10 años de narrativas en el TOCCA - saberes y prácticas transdisciplinarios entre las artes y la salud

Este texto tem como objetivo apresentar as experiências do "Programa TOCCA (Terapia Ocupacional, Corpo, Cultura e Artes): saberes e práticas transdisciplinares entre as artes e a saúde" na Universidade Federal de Santa Maria, que procura conjugar arte e clínica em proposições poéticas que promovam a produção da vida. Trata-se de uma narrativa reflexiva do programa em seus dez anos de atividades de ensino, pesquisa e extensão, buscando problematizar os saberes e práticas compartilhados nos trajetos realizados. Acredita-se ser esse um importante caminho para rever como prosseguir, cuidando para ativar a cooperação e a partilha das diferenças desejadas e vividas. (AU)

This paper presents the experiences of the "TOCCA Program: Transdisciplinary Knowledge and Practices in Arts and Health" at Santa Maria Federal University. The program brings together arts and clinical practice in poetic propositions that promote the production of life. This reflective narrative of the program and its 10 years of teaching, research and extension activities aims to problematize the knowledge and practices shared over the course of the program. We believe this exercise is an important way of reviewing how to proceed, taking care to activate cooperation and share desired and lived differences. (AU)

Este texto tiene el objetivo de presentar las experiencias del "Programa TOCCA: Saberes y prácticas transdisciplinarios entre las artes y la salud" en la Universidad Federal de Santa Maria que busca conjugar arte y clínica en propuestas poéticas que promuevan la producción de la vida. Se trata de una narrativa reflexiva del programa en sus diez años de actividades de enseñanza, investigación y extensión, buscando problematizar los saberes y prácticas compartidos en los trayectos realizados. Se cree que este es un importante camino para revisar cómo proseguir, cuidando para activar la cooperación y compartición de las diferencias deseadas y vividas. (AU)

Unconventional amino acids in medicinal chemistry: First report on taurine merged within carbonic anhydrase inhibitors.

This study reports the design, synthesis of a series of taurine containing benzenesulfonamide derivatives which were all screened in vitro against the physiological relevant human (h) expressed Carbonic Anhydrase (CA; EC 4.2.1.1) I, II, IX, XII isozymes. Compound 2, 5, 11-16 displayed superior inhibitory activities against the tumor associated hCA IX over the reference drug Acetazolamide (AAZ). Both hCA IX and XII isoforms were selectively inhibited only by compound 3, whereas the chloro-containing compound 12 was showed as the most selective and effective inhibitor profile for the CA IX isoforms. To the best of our knowledge the data reported herein are the first of this kind and introduce in the literature new compounds worth for future development within the Medicinal Chemistry field.

Screening of benzenesulfonamide in combination with chemically diverse fragments against carbonic anhydrase by differential scanning fluorimetry.

The differential scanning fluorimetry (DSF) screening of 5.692 fragments in combination with benzenesulfonamide (BSA) against bovine carbonic anhydrase (bCA) delivered >100 hits that either caused, on their own, a significant thermal shift (ΔTm, °C) in the protein melting temperature or significantly influenced the thermal shift observed for BSA alone. Three hits based on 1,2,3-triazole moiety represent the periphery of the recently reported potent inhibitors of hCA II, IX and XII which were efficacious in vivo. Such a re-discovery of suitable BSA periphery essentially validates the new fragment-based approach to the discovery of future CAIs. Structures of other validated fragment hits are reported.

A structure-based approach towards the identification of novel antichagasic compounds: Trypanosoma cruzi carbonic anhydrase inhibitors.

Trypanosoma cruzi carbonic anhydrase (TcCA) has recently emerged as an interesting target for the design of new compounds to treat Chagas disease. In this study we report the results of a structure-based virtual screening campaign to identify novel and selective TcCA inhibitors. The combination of properly validated computational methodologies such as comparative modelling, molecular dynamics and docking simulations allowed us to find high potency hits, with KI values in the nanomolar range. The compounds also showed trypanocidal effects against T. cruzi epimastigotes and trypomastigotes. All the candidates are selective for inhibiting TcCA over the human isoform CA II, which is encouraging in terms of possible therapeutic safety and efficacy.

Further validation of strecker-type α-aminonitriles as a new class of potent human carbonic anhydrase II inhibitors: hit expansion within the public domain using differential scanning fluorimetry leads to chemotype refinement.

Testing of an expanded, 800-compound set of analogues of the earlier described Strecker-type α-aminonitriles (selected from publicly available Enamine Ltd. Screening Collection) in thermal shift assay against bovine carbonic anhydrase (bCA) led to further validation of this new class of inhibitors and identification a new, refined chemotype represented by inhibitors with 10-improved potency. [Formula: see text].

Improvement of sea fennel (Crithmum maritimum L.) nutritional value through iodine biofortification in a hydroponic floating system.

Sea fennel is an herbaceous aromatic and edible halophyte, naturally occurring in coastal areas of the Mediterranean basin. Besides its scientific interest as a salt-tolerant species it exhibits considerable nutritional value and economical potential. As sea fennel is distributed in maritime areas, where natural iodine is available in high concentrations, the aim of this study was to evaluate whether sea fennel has the potential to accumulate elevated iodine concentrations under cultivation. A biofortification experiment in a hydroponic system applying two forms of iodine, KI and KIO3 in different concentrations was set up and monitored using sophisticated targeted elementary and metabolite analysis. The biofortification potential and possible effects on nutritional value were investigated. This study revealed that both iodine forms increased the iodine content of sea fennel tissues. Nutritional and health promoting components, biomass production but also antioxidant potential were stable or even improved under the iodine treatments.

Syntesis of thio- and seleno-acetamides bearing benzenesulfonamide as potent inhibitors of human carbonic anhydrase II and XII.

A novel series of thio- and seleno-acetamides bearing benzenesulfonamide were synthetized and tested as human carbonic anhydrase inhibitors. These compounds were tested for the inhibition of four human (h) isoforms, hCA I, II, IX, and XII, involved in pathologies such as glaucoma (CA II and XII) or cancer (CA IX/XII). Several derivatives showed potent inhibition activity in low nanomolar range such as 3a, 4a, 7a and 8a. Furthermore, based on the tail approach we explain the interesting and selective inhibition profile of compound such as 5a and 9a, which were more selective for hCA I, 9b which was selective for hCA II, 3f selective for hCA IX and finally, 3e and 4b selective for hCA XII, over the other three isoforms. They are interesting leads for the development of more effective and isoform-selective inhibitors.

Sulfur, selenium and tellurium containing amines act as effective carbonic anhydrase activators.

A new series of ß-aminochalcogenides were designed and synthesized to identify new carbonic anhydrase activator (CAA) agents as novel tools for the management of several neurodegenerative and metabolic disorders which represent a clinical challenge without effective therapies available. Some ß-aminoselenides and ß-aminotellurides showed effective CA activating effects and a potent antioxidant activity. CAAs may have applications for memory therapy and CA deficiency syndromes.

Bioactive isoflavones from Pueraria lobata root and starch: Different extraction techniques and carbonic anhydrase inhibition.

Kudzu, the dried root of an important edible plant (Pueraria lobata), is used in Traditional Chinese Medicine for the important nutritional value strictly related to its isoflavone derivatives. These compounds characterize the quality of kudzu contained in different preparations, as pharmaceutical ingredient as well as dietary/food supplement (e.g. starch). The optimization of the isoflavones recovery, monitored by HPLC-PDA, through different innovative and conventional extraction techniques, e.g. microwave-assisted, ultrasound-assisted and conventional extraction, represented a suitable challenge in food industry and natural products evaluation. The impact on the isoflavone extraction by using an ionic liquid-assisted procedure was also considered. Furthermore, the inhibitory activity of the most representative isoflavones, isolated from kudzu, was evaluated using four isoforms (I, II, IX and XII) of human carbonic anhydrase (hCA) due to their role in several physiopathological processes.

Acyl selenoureido benzensulfonamides show potent inhibitory activity against carbonic anhydrases from the pathogenic bacterium Vibrio cholerae.

A series of acyl selenoureido benzensulfonamides was evaluated as carbonic anhydrase (CA, EC 4.2.1.1) inhibitors against two Vibrio cholerae such enzymes (VchCAα over VchCAß) belonging to the α- and ß-classes, potential novel targets for anti-infective drugs development. These compounds showed strong inhibitory action against VchCAα over VchCAß and excellent selectivity over the human (h) off-target isoforms hCA I and II. Identification of potent and possibly selective inhibitors of VchCAα and/or VchCAß over the human counterparts may lead to pharmacological tools useful for understanding the physiological role(s) of these under-investigated proteins, possibly involved in the virulence of the bacterium and colonization of the host in bicarbonate rich regions of the gastro-intestinal tract.

Microwave-assisted extraction, HPLC analysis, and inhibitory effects on carbonic anhydrase I, II, VA, and VII isoforms of 14 blueberry Italian cultivars.

The multi-component fingerprint and the biological evaluation of plant-derived material are indispensable for the pharmaceutical field, in food quality control procedures, and in all plant-based products. We investigated the quantitative content of biologically active compounds (anthocyanins and chlorogenic acid) of microwave-assisted blueberry extracts from 14 different Italian cultivars, using validated high-performance liquid chromatography-photodiode array detector (HPLC-PDA) method and routinely instrument configuration. The carbonic anhydrase (CA, EC 4.2.1.1) inhibition profiles against several pharmacologically relevant CA isoforms of blueberry extracts and some bioactive compounds were also investigated. The various cultivars showed a highly variable content in anthocyanins and chlorogenic acid, and their CA inhibitory effects were also highly variable. Overall these data prove that antioxidant natural products found in blueberries may be useful for designing pharmacological agents in which various CAs are involved, e.g., antiobesity, antitumor, or anticonvulsants agents.

Formação em terapia ocupacional na interface das artes e da saúde: a experiência do PACTO / Education in occupational therapy in the art & health interface: the PACTO experience

Este artigo apresenta o trabalho de formação oferecido na Graduação em Terapia Ocupacional desenvolvido pelo Laboratório de Estudos e Pesquisa Arte, Corpo e Terapia Ocupacional da FMUSP. O Laboratório realiza atividades de ensino e pesquisa, no campo didático-assistencial, nos atendimentos grupais e individuais para populações heterogêneas através do Programa Permanente Composições Artísticas e Terapia Ocupacional (PACTO) e em parcerias com projetos que se constituem na interface arte-saúde. As experiências dos estudantes são acolhidas e elaboradas em discussões de equipe, supervisões semanais, grupo de estudos e seminários de aprofundamento temático. A formação oferecida habilita os estudantes para: a compreensão e a construção de práticas na interface arte/saúde; a problematização das abordagens na interface arte/saúde nos espaços de assistência em saúde e nos espaços culturais; a compreensão das relações institucionais e sociais e dos processos de transformação das instituições; a reflexão sobre a participação do terapeuta ocupacional nas políticas culturais e nas políticas de humanização em saúde. Elegemos relatar e pontuar aspectos do trabalho efetuado nos últimos anos (2003-2008) no qual algumas estratégias de ensino foram consolidadas e evidenciaram seus efeitos na formação dos estudantes e no exercício profissional. No campo da Terapia Ocupacional verifica-se uma ampliação das possibilidades de atuação profissional vinculadas ao campo da produção cultural, em projetos transdisciplinares voltados para construção de redes sociais inclusivas, com a participação coletiva e a produção de subjetividade.

This paper presents the education work offered in the Occupational Therapy Program developed by the Laboratory of Studies and Research in Art, Body and Occupational Therapy of FMUSP. The Laboratory conducts teaching and research activities, in the didactic-assistancial field, with groups and individuals accompaniment for heterogeneous population through the Artistic Compositions and Occupational Therapy Permanent Program (PACTO) and through partnerships made with projects in the art/health interface. Students experiences are sheltered and elaborated in team discussion, weekly supervisions, study groups and theme-deepening seminars. The education qualifies students to: understand and build up practices in the art/health interface under an occupational therapy's perspective; questioning the approaches in the art/health interface in healthcare spaces and in cultural spaces; comprehend social and institutional relations and institutional transformation processes; understanding the occupational therapists' participation in cultural policies and health humanizing policies. We chose to report and scoring aspects of the work done in recent years (2003-2008) in which some teaching strategies were consolidated and showed its effects on students' education and professional exercise. In the Occupational Therapy field there is an increase in the possibilities for professional performance linked to the issues of cultural production, in trans-disciplinary projects dealing with constructing inclusive social networks, with collective participation and production of subjectivity.

A criação de um espaço para a existência: o Espaço Lúdico Terapêutico / The creation of space to existence: therapeutic playful space

O artigo apresenta o percurso realizado pelo Espaço Lúdico Terapêutico, serviço de assistência à criança e adolescente com deficiência mental e distúrbios globais do desenvolvimento do Laboratório de Estudos sobre Deficiência e Cotidiano do Curso de Terapia Ocupacional da USP, desde a sua criação em 1996 até os dias de hoje / This article gives an account of the path carried out by the 'Therapeutic Playful Space' an assistance service to children and adolescents mentally handicapped and with and with global disorders in their development. In its Laboratory of Studies on Deficiency and Daily Routine in the Occupational Therapy Course much has been...