Diseño y estudio de compuestos de rutenio con actividad biológica

Programa Oficial de Doutoramento en Química Ambiental e Fundamental. 5031V01[Resumo] O presente proxecto aborda a síntese e caracterización estructural de novos compostos de coordinación mono e polinucleares de rutenio derivados de bipiridina e novos compostos organometálicos mononucleares de rutenio derivados de p-cimeno, e un estudo das propiedades citotóxicas y toxicoloxía in vivo co fin de determinar a súa aplicabilidade como axentes antitumorais. A determinación estrutural levouse a cabo mediante análise elemental, espectrometría de masas, RMN de 1H y 31P-{1H}, espectroscopia de IR e UV-Vis, medidas de condutividade, voltametría cíclica e difracción de rayos X de monocristal. Estudiouse a actividade citotóxica dos compostos en distintas liñas de cancro humano, a interacción co ADN como molécula diana e a interacción coa albúmina como sistema de transporte, e o mecanismo de morte celular (apoptose e/ou autofaxia). Levouse a cabo o análise da toxicidade in vivo dos compostos utilizando o modelo de embrión de pez cebra. A finalidade global dos estudos realizados permitiu establecer as relacións entre a estrutura e o comportamento dos compostos como axentes antineoplásicos. Os resultados indican que moitos compostos presentan actividade citotóxica similar e incluso superior ó cis-platino, con una mínima citotoxicidade en células non tumorais e con baixa ou ningunha letalidade no modelo de pez cebra.[Abstract] This project is based on the synthesis and structural characterization of new mono- and polinuclear coordination compounds of ruthenium derived from bipyridine and new mononuclear organometallic compounds of ruthenium derived from pcymene, and the study of their cytotoxic properties and in vivo toxicology in order to determine their applicability as antitumor agents. The structural determination has been carrie out by elemental analysis, mass spectrometry, 1H and 31P-{1H} NMR, UV-Vis and IR spectroscopy, conductivity measurements, cyclic voltammetry and single-crystal X-Ray difraction. It was evaluated the cytotoxic activity in human cancer cell lines, the interaction with DNA as target molecule and the interaction with albumin as transport system were evaluated. The in vivo toxicology was performed using the zebrafish model. The overall goal of the studies allowed to stablish the relationship between the structure and the behaviour of the compounds as antineoplastic agents. The results obtained results indicate that the compounds show a similar or even higher cytotoxic activity compared with cis-platinum, a minimum cytotoxicity against non-tumour cells as well as low or no lethality in the zebrafish model.[Resumen] La temática del presente proyecto aborda la síntesis y caracterización de nuevos compuestos mono- y polinucleares de rutenio derivados de bipiridina y nuevos compuestos de rutenio derivados de p-cimeno, para posteriormente estudiar sus aptitudes corno agentes antitumorales_ Con este fin, se estudió la actividad citotóxica de los compuestos en distintas líneas de cáncer humano, cuyos resultados permitieron determinar que perfil estructural permitía una mejora de la actividad_ Como posible forma de actuación de los compuestos sintetizados se realizó un estudio de interacción con el ADN corno posible diana biológica mediante la espectroscopia de ultravioleta visible y la espectroscopia de dicroísmo circular_ Corno posible vía de transporte se estudió la interacción con la albúmina humana_ Los estudios de interacción de los compuestos con el ADN y con la albúmina humana fueron contrastados mediante modelos teóricos de interacción, empleando las estructuras obtenidas a través de difracción de rayos x de monocristal y mediante el cálculo de la optimización estructural de los compuestos_ Para profundizar en el mecanismo de acción de los compuestos se estudió la muerte celular llegándose a la conclusión de que los compuestos estudiados inducen tanto la apoptosis corno la autofagia_ Como el estudio de la toxicidad en modelos in vivo es clave en el desarrollo de fármacos se estudió la toxicidad de los compuestos in vivo en el modelo pez cebra_ La finalidad global de los estudios realizados permitió relacionar la estructura con la actividad de los compuestos y ratificar que el rutenio se establece como candidato alternativo al platino en el tratamiento del cáncer- Así, se han obtenido estructuras que presentan actividad citotóxica similar e incluso superior al Gis-platino con apenas citotoxicidad en células no tumorales y con baja o ninguna letalidad en el modelo de pez cebra

Marine Natural Products from the Yucatan Peninsula

[Abstract] Mexico is one of the three areas of the world with the greatest terrestrial and cultural biological diversity. The diversity of Mexican medicinal flora has been studied for a long time and several bioactive compounds have been isolated. The investigation of marine resources, and particularly the potential of Mexican marine resources, has not been intensively investigated, even though the Yucatan Peninsula occupies 17.4% of the total of the Mexican coast, with great biological diversity in its coasts and the ocean. There are very few studies on the chemistry of natural products from marine organisms that were collected along the coasts of the Yucatan Peninsula and most of them are limited to the evaluation of the biological activity of their organic extracts. The investigations carried out on marine species from the Yucatan Peninsula resulted in the identification of a wide structural variety of natural products that include polyketides, terpenoids, nitrogen compounds, and biopolymers with cytotoxic, antibacterial, antifouling, and neurotoxic activities. This review describes the literature of bioprospecting and the exploration of the natural product diversity of marine organisms from the coasts of the Yucatan Peninsula up to mid-2019.This work was supported by grants RTI2018-093634-B-C22 (AEI/FEDER, EU) from the State Agency for Research (AEI) of Spain, co-funded by the FEDER Programme from the European Union, and GRC2018/039 and ED431E 2018/03 of CICA-INIBIC strategic group from Xunta de Galicia. D.P.P. received a fellowship from the program National Council of Science and Technology (CONACYT) of Mexico and the Secretariat of Research, Innovation and Higher Education (SIIES) of Yucatan (Mexico). O.A.L.R. received a financial support from MostMicro unit. Project LISBOA-01-0145-FEDER-007660 of PortugalXunta de Galicia; GRC2018/039Xunta de Galicia; ED431E 2018/03Portugal. Fundação para a Ciência e a Tecnologia; LISBOA-01-0145-FEDER-00766

In Vitro and In Vivo Biological Activity of Ruthenium 1,10-Phenanthroline-5,6-dione Arene Complexes

Funding Information: We are grateful to Fundação para a Ciência e a Tecnologia, I.P., through MOSTMICRO-ITQB R&D Unit (UIDB/04612/2020, UIDP/04612/2020) and LS4FUTURE Associated Laboratory (LA/P/0087/2020). The NMR spectrometers at CERMAX are integrated in the national NMR Network and partially supported through project 022162. Oscar A. Lenis-Rojas acknowledge national funds through FCT, POPH-Programa Operacional Potencial Humano, and FSE (European Social Fund) for the CEEC 2017 Initiative. Additionally, this work is financed by national funds from FCT—Fundação para a Ciência e a Tecnologia, I.P., in the scope of the project UIDP/04378/2020 and UIDB/04378/2020 of the Research Unit on Applied Molecular Biosciences—UCIBIO and the project LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy—i4HB. Publisher Copyright: © 2022 by the authors.Ruthenium(II) arene complexes exhibit promising chemotherapeutic properties. In this study, the effect of the counter anion in Ru(II) complexes was evaluated by analyzing the biological effect of two Ru(II) p-cymene derivatives with the 1,10-phenanthroline-5,6-dione ligand of general-formula [(η6-arene)Ru(L)Cl][X] X = CF3SO3 (JHOR10) and PF6 (JHOR11). The biological activity of JHOR10 and JHOR11 was examined in the ovarian carcinoma cell line A2780, colorectal carcinoma cell line HCT116, doxorubicin-resistant HCT116 (HCT116-Dox) and in normal human dermal fibroblasts. Both complexes JHOR10 and JHOR11 displayed an antiproliferative effect on A2780 and HCT116 cell lines, and low cytotoxicity in fibroblasts. Interestingly, JHOR11 also showed antiproliferative activity in the HCT116-Dox cancer cell line, while JHOR10 was inactive. Studies in A2780 cells showed that JHOR11 induced the production of reactive oxygen species (ROS) that trigger autophagy and cellular senescence, but no apoptosis induction. Further analysis showed that JHOR11 presented no tumorigenicity, with no effect in the cellular mobility, as evaluated by thye wound scratch assay, and no anti- or pro-angiogenic effect, as evaluated by the ex-ovo chorioallantoic membrane (CAM) assay. Importantly, JHOR11 presented no toxicity in chicken and zebrafish embryos and reduced in vivo the proliferation of HCT116 injected into zebrafish embryos. These results show that these are suitable complexes for clinical applications with improved tumor cell cytotoxicity and low toxicity, and that counter-anion alteration might be a viable clinical strategy for improving chemotherapy outcomes in multidrug-resistant (MDR) tumors.publishersversionpublishe

Evaluation of the in vitro and in vivo efficacy of ruthenium polypyridyl compounds against breast cancer

[Abstract] The clinical success of cisplatin, carboplatin, and oxaliplatin has sparked the interest of medicinal inorganic chemistry to synthesize and study compounds with non-platinum metal centers. Despite Ru(II)–polypyridyl complexes being widely studied and well established for their antitumor properties, there are not enough in vivo studies to establish the potentiality of this type of compound. Therefore, we report to the best of our knowledge the first in vivo study of Ru(II)–polypyridyl complexes against breast cancer with promising results. In order to conduct our study, we used MCF7 zebrafish xenografts and ruthenium complexes [Ru(bipy)2(C12H8N6-N,N)][CF3SO3]2Ru1 and [{Ru(bipy)2}2(μ-C12H8N6-N,N)][CF3SO3]4Ru2, which were recently developed by our group. Ru1 and Ru2 reduced the tumor size by an average of 30% without causing significant signs of lethality when administered at low doses of 1.25 mg·L−1. Moreover, the in vitro selectivity results were confirmed in vivo against MCF7 breast cancer cells. Surprisingly, this work suggests that both the mono- and the dinuclear Ru(II)–polypyridyl compounds have in vivo potential against breast cancer, since there were no significant differences between both treatments, highlighting Ru1 and Ru2 as promising chemotherapy agents in breast cancer therapy.Xunta de Galicia; ED431C 2018/39Portugal. Fundação para a Ciência e a Tecnologia; PEst 2015-2020Portugal. Fundação para a Ciência e a Tecnologia; UID/Multi/04349/2013Portugal. Fundação para a Ciência e a Tecnologia; RECI/QEQ-QIN/0189/2012Portugal. Fundação para a Ciência e a Tecnologia; UID/QUI/00100/2020Portugal. Fundação para a Ciência e a Tecnologia; UIDP/04378/2020Portugal. Fundação para a Ciência e a Tecnologia; UIDB/04378/2020Portugal. Fundação para a Ciência e a Tecnologia; LA/P/0140/202

<i>N</i>-Heterocyclic Carbene Iron Complexes as Anticancer Agents: In Vitro and In Vivo Biological Studies

Cisplatin and its derivatives are commonly used in chemotherapeutic treatments of cancer, even though they suffer from many toxic side effects. The problems that emerge from the use of these metal compounds led to the search for new complexes capable to overcome the toxic side effects. Here, we report the evaluation of the antiproliferative activity of Fe(II) cyclopentadienyl complexes bearing n-heterocyclic carbene ligands in tumour cells and their in vivo toxicological profile. The in vitro antiproliferative assays demonstrated that complex Fe1 displays the highest cytotoxic activity both in human colorectal carcinoma cells (HCT116) and ovarian carcinoma cells (A2780) with IC50 values in the low micromolar range. The antiproliferative effect of Fe1 was even higher than cisplatin. Interestingly, Fe1 showed low in vivo toxicity, and in vivo analyses of Fe1 and Fe2 compounds using colorectal HCT116 zebrafish xenograft showed that both reduce the proliferation of human HCT116 colorectal cancer cells in vivo

Ru-II(p‑cymene) compounds as effective and selective anticancer candidates with no toxicity in vivo

Ruthenium(II) complexes are currently considered a viable alternative to the widely used platinum complexes as efficient anticancer agents. We herein present the synthesis and characterization of half-sandwich ruthenium compounds with the general formula [Ru(p-cymene)(L-N,N)Cl][CF3SO3] (L = 3,6-di-2-pyridyl-1,2,4,5-tetrazine (1) 6,7-dimethyl-2,3-bis(pyridin-2-yl)quinoxaline (2)), which have been synthesized by substitution reactions from the precursor dimer [Ru(p-cymene)(Cl)(μ-Cl)]2 and were characterized by elemental analysis, mass spectrometry, 1H NMR, UV–vis, and IR spectroscopy, conductivity measurements, and cyclic voltammetry. The molecular structure for complex 2 was determined by single-crystal X-ray diffraction. The cytotoxic activity of these compounds was evaluated against human tumor cells, namely ovarian carcinoma A2780 and breast MCF7 and MDAMB231 adenocarcinoma cells, and against normal primary fibroblasts. Whereas the cytotoxic activity of 1 is moderate, IC50 values found for 2 are among the lowest previously reported for Ru(p-cymene) complexes. Both compounds present no cytotoxic effect in normal human primary fibroblasts when they are used at the IC50 concentration in A2780 and MCF7 cancer cells. Their antiproliferative capacity is associated with a combined mechanism of apoptosis and autophagy. A strong interaction with DNA was observed for both with a binding constant value of the same magnitude as that of the classical intercalator [Ru(phen)2(dppz)]2+. Both complexes bind to human serum albumin with moderate to strong affinity, with conditional binding constants (log Kb) of 4.88 for complex 2 and 5.18 for complex 1 in 2% DMSO/10 mM Hepes pH7.0 medium. The acute toxicity was evaluated in zebrafish embryo model using the fish embryo acute toxicity test (FET). Remarkably, our results show that compounds 1 and 2 are not toxic/lethal even at extremely high concentrations. The novel compounds reported herein are highly relevant antitumor metallodrug candidates, given their in vitro cytotoxicity toward cancer cells and the lack of in vivo toxicity.info:eu-repo/semantics/publishedVersio

Antifungal Potential of Marine Organisms of the Yucatan Peninsula (Mexico) against Medically Important <i>Candida</i> spp.

Invasive fungal infections represent a global health threat. They are associated with high mortality and morbidity rates, partly due to the ineffectiveness of the available antifungal agents. The rampant increase in infections recalcitrant to the current antifungals has worsened this scenario and made the discovery of new and more effective antifungals a pressing health issue. In this study, 65 extracts from marine organisms of the Yucatan Peninsula, Mexico, were screened for antifungal activity against Candida albicans and Candida glabrata, two of the most prevalent fungal species that cause nosocomial invasive fungal infections worldwide. A total of 51 sponges, 13 ascidians and 1 gorgonian were collected from the coral reef and mangrove forest in the Yucatan Peninsula (Mexico) and extracted with organic solvents. Nine crude extracts showed potent antifungal activity, of which four extracts from the sponge species Aiolochroia crassa, Amphimedon compressa, Monanchora arbuscula and Agelas citrina had promising activity against Candida spp. Bioassay-guided fractionation of the M. arbuscula extract revealed the remarkable fungicidal activity of some fractions. Analysis of the chemical composition of one of the most active fractions by UHPLC-HRMS and NMR indicated the presence of mirabilin B and penaresidin B, and their contribution to the observed antifungal activity is discussed. Overall, this work highlights marine organisms of the Yucatan Peninsula as important reservoirs of natural products with promising fungicidal activity, which may greatly advance the treatment of invasive fungal infections, especially those afflicting immunosuppressed patients

Half-sandwich Ru(p-cymene) compounds with diphosphanes: In Vitro and In Vivo evaluation as potential anticancer metallodrugs

Ruthenium(II) complexes are currently considered attractive alternatives to the widely used platinum-based drugs. We present herein the synthesis and characterization of half-sandwich ruthenium compounds formulated as [Ru(p-cymene)(L)Cl]-[CF3SO3] (L = 1,1-bis(methylenediphenylphosphano)ethylene, 1; L = 1,1-bis(diphenylphosphano)ethylene, 2), which were characterized by elemental analysis, mass spectrometry, H-1 and P-31{H-1} NMR, UV-vis and IR spectroscopy, conductivity measurements and cyclic voltammetry. The molecular structures for both complexes were determined by single-crystal X-ray diffraction. Their cytotoxic activity was evaluated using the MTT assay against human tumor cells, namely ovarian (A2780) and breast (MCF7 and MDA-MB-231). Both complexes were active against breast adenocarcinoma cells, with complex 1 exhibiting a quite remarkable cytotoxicity in the submicromolar range. Interestingly, at concentrations equivalent to the IC50 values in the MCF7 cancer cells, complexes 1 and 2 presented lower cytotoxicity in normal human primary fibroblasts. The antiproliferative effects of 1 and 2 in MCF7 cells might be associated with the induction of reactive oxygen species (ROS), leading to a combined cell death mechanism via apoptosis and autophagy. Despite the fact that in vitro a partial intercalation between complexes and DNA was observed, no MCF7 cell cycle delay or arrest was observed, indicating that DNA might not be a direct target. Complexes 1 and 2 both exhibited a moderate to strong interaction with human serum albumin, suggesting that protein targets may be involved in their mode of action. Their acute toxicity was evaluated in the zebrafish model. Complex 1 (the most toxic of the two) exhibited a lethal toxicity LC50 value about 1 order of magnitude higher than any IC50 concentrations found for the cancer cell models used, highlighting its therapeutic relevance as a drug candidate in cancer chemotherapy.info:eu-repo/semantics/publishedVersio

Dinuclear Ru^II(bipy)₂ Derivatives: Structural, Biological, and in Vivo Zebrafish Toxicity Evaluation

Ruthenium-based drugs exhibit interesting properties as potential anticancer pharmaceuticals. We herein present the synthesis and characterization of a new family of ruthenium complexes with formulas [{Ru­(bipy)₂}₂(<i>μ-</i>L)]­[CF₃SO₃]₄ (L = bptz, <b>1a</b>) and [{Ru­(bipy)₂}₂(<i>μ-</i>L)]­[CF₃SO₃]₂ (L = arphos, <b>2a</b>; dppb, <b>3a</b>; dppf, <b>4a</b>), which were synthesized from the Ru­(II) precursor compound <i>cis</i>-Ru­(bipy)₂Cl₂. The complexes were characterized by elemental analysis, mass spectrometry, ¹H and ³¹P­{¹H} NMR, IR spectroscopy, and conductivity measurements. The molecular structures for three Ru­(II) compounds were determined by single-crystal X-ray diffraction. The newly developed compounds interact with CT-DNA by intercalation, in particular, <b>2a</b>, <b>3a</b>, and <b>4a</b>, which also seemed to induce some extent of DNA degradation. This effect seemed to be related with the formation of reactive oxygen species. The cytotoxic activity was evaluated against A2780, MCF7, and MDAMB231 human tumor cells. Compounds <b>2a</b> and <b>4a</b> were the most cytotoxic with activity compared to cisplatin (∼2 μM, 72 h) in the A2780 cisplatin sensitive cells. All the compounds induced A2780 cell death by apoptosis, however, to a lesser extent for compounds <b>4a</b> and <b>2a</b>. For these compounds, the mechanism of cell death in addition to apoptosis seemed to involve autophagy. In vivo toxicity was evaluated using the zebrafish embryo model. LC₅₀ estimates varied from 5.397 (<b>3a</b>) to 39.404 (<b>1a</b>) mg/L. Considering the in vivo toxicity in zebrafish embryos and the in vitro cytotoxicity in cancer cells, compound <b>1a</b> seems to be the safest having no effect on dechirionation and presenting a good antiproliferative activity against ovarian carcinoma cells