Diclofenac N-Derivatives as Therapeutic Agents with Anti-Inflammatory and Anti-Cancer Effect

A series of diclofenac N-derivatives (2, 4, 6, 8c, 9c, 10a-c) were synthesized in order to test their anti-cancer and anti-inflammatory effects. The anticarcinogen activity has been assayed against three cancer cell lines: HT29, human colon cancer cells; Hep-G2, human hepatic cells; and B16-F10, murine melanoma cells. First, we determined the cytotoxicity of the different compounds, finding that the most effective compound was compound 8c against all cell lines and both compounds 4 and 6 in human Hep-G2 and HT29 cell lines. Compounds 4 and 8c were selected for the percentage of apoptosis determination, cell cycle distribution, and mitochondrial membrane potential measure because these products presented the lowest IC50 values in two of the three cancer cell lines assayed (B16-F10 and HepG2), and were two of the three products with lowest IC50 in HT29 cell line. Moreover, the percentages of apoptosis induction were determined for compounds 4 and 8c, showing that the highest values were between 30 to 60%. Next, the effects of these two compounds were observed on the cellular cycle, resulting in an increase in the cell population in G2/M cell cycle phase after treatment with product 8c, whereas compound 4 increased the cells in phase G0/G1, by possible differentiation process induction. Finally, to determine the possible apoptosis mechanism triggered by these compounds, mitochondrial potential was evaluated, indicating the possible activation of extrinsic apoptotic mechanism. On the other hand, we studied the anti-inflammatory effects of these diclofenac (DCF) derivatives on lipopolysaccharide (LPS) activated RAW 264.7 macrophagesmonocytes murine cells by inhibition of nitric oxide (NO) production. As a first step, we determined the cytotoxicity of the synthesized compounds, as well as DCF, against these cells. Then, sub-cytotoxic concentrations were used to determine NO release at different incubation times. The greatest antiinflammatory effect was observed for products 2, 4, 8c, 10a, 10b, and 9c at 20 µg·mL−1 concentration after 48 h of treatment, with inhibition of produced NO between 60 to 75%, and a concentration that reduces to the 50% the production of NO (IC50 NO) between 2.5 to 25 times lower than that of DCF. In this work, we synthesized and determined for the first time the anti-cancer and anti-inflammatory potential of eight diclofenac N-derivatives. In agreement with the recent evidences suggesting that inflammation may contribute to all states of tumorigenesis, the development of these new derivatives capable of inducing apoptosis and anti-inflammatory effects at very low concentrations represent new effective therapeutic strategies against these diseases.MINISTERIO DE ECONOMÍA Y COMPETITIVIDAD, PID2019-106222RB-C32/SRA (State Research Agency, 10.13039/501100011033)“Consejería de Economía, Conocimiento, Empresas y Universidad. Junta de Andalucía”, grant number B1-BIO-281-UGR1

Exosome: A New Player in Translational Nanomedicine

This research was funded by the Spanish ISCIII Health Research Fund and the European Regional Development Fund (FEDER) through research grants PI12/01097, PI15/02015, PI18/00337 (F.M.), PIE16-00045 (J.A.M.), DTS19/00145 (J.A.M.) and PI18/00330 (K.B.), as well as by the Spanish Ministry of Science, Innovation and Universities (MICIU) through FEDER research grant RTI2018-101309-B-C2 (J.A.M.). The CECEyU and CSyF Councils of the Junta de Andalucia FEDER/European Cohesion Fund (FSE) provided the following research grants: 2016000073391-TRA, 2016000073332-TRA, PI-57069 and PAIDI-Bio326 (F.M.) and PI-0014-2016 (K.B). K.B. was also on a Nicolas Monardes Regional Ministry of Health contract (0006/2018). H.A. is supported by Research Excellence PhD Fellowship (2UAE2020) from the National Center for Scientific and Technical Research (CNRST). M.T.-M. is funded by the Spanish Ministry of Science and Innovation (SMSI) through an FPU16/05467 fellowship. M.C.-G. is funded by SMSI through a GJ fellowship (PEJ-2018-001760-A).We wish to thank Michael O’Shea for proofreading the review. Figures were created using Biorender.com.Summary: Exosomes are extracellular vesicles released by the vast majority of cell types both in vivo and ex vivo, upon the fusion of multivesicular bodies (MVBs) with the cellular plasma membrane. Two main functions have been attributed to exosomes: their capacity to transport proteins, lipids and nucleic acids between cells and organs, as well as their potential to act as natural intercellular communicators in normal biological processes and in pathologies. From a clinical perspective, the majority of applications use exosomes as biomarkers of disease. A new approach uses exosomes as biologically active carriers to provide a platform for the enhanced delivery of cargo in vivo. One of the major limitations in developing exosome-based therapies is the difficulty of producing sufficient amounts of safe and efficient exosomes. The identification of potential proteins involved in exosome biogenesis is expected to directly cause a deliberate increase in exosome production. In this review, we summarize the current state of knowledge regarding exosomes, with particular emphasis on their structural features, biosynthesis pathways, production techniques and potential clinical applications.Spanish ISCIII Health Research Fund PI12/01097 PI15/02015 PI18/00337 PIE16-00045 DTS19/00145 PI18/00330European Union (EU) PI12/01097 PI15/02015 PI18/00337 PIE16-00045 DTS19/00145 PI18/00330Spanish Ministry of Science, Innovation and Universities (MICIU) through FEDER research grant RTI2018-101309-B-C2CECEyU Council of the Junta de Andalucia FEDER/European Cohesion Fund (FSE) 2016000073391-TRA 2016000073332-TRA PI-57069 PAIDI-Bio326 PI-0014-2016CSyF Council of the Junta de Andalucia FEDER/European Cohesion Fund (FSE) 2016000073391-TRA 2016000073332-TRA PI-57069 PAIDI-Bio326 PI-0014-2016Nicolas Monardes Regional Ministry of Health contract 0006/2018Research Excellence PhD Fellowship from the National Center for Scientific and Technical Research (CNRST) 2UAE2020Spanish Government FPU16/05467SMSI through a GJ fellowship PEJ-2018-001760-

Exosomes: Their Role in Pathogenesis, Diagnosis and Treatment of Diseases

This study was funded by the Spanish ISCIII Health Research Fund and the European Regional Development Fund (FEDER) through research grants PI12/01097, PI15/02015, PI18/00337 (F.M.), PIE16-00045 (J.A.M.), DTS19/00145 (J.A.M.) and PI18/00330 (K.B.). The study was also supported by the Ministry of Science, Innovation and Universities (MICIU, RTI2018-101309-B-C22, FEDER funds), by the Consejeria de Economia, Conocimiento, Empresas y Universidad de la Junta de Andalucia (SOMM17/6109/UGR, FEDER Funds), the Chair of "Doctors Galera-Requena in cancer stem cell research" (CMC-CTS963) and the Junta de Andalucia Health and Families Department (CARTPI-0001-201). The CECEyU and CSyF of the Junta de Andalucia FEDER/European Cohesion Fund (FSE) provided the following research grants: 2016000073391-TRA, 2016000073332-TRA, PI57069 and PAIDI-Bio326 (F.M.) and PI-0014-2016 (K.B.). K.B. was also on a Nicolas Monardes Regional Ministry of Health contract (0006/2018). H.A. held a Research Excellence PhD Fellowship (2UAE2020) from the National Center for Scientific and Technical Research (CNRST). L.M. was supported by the Mohammed VI Polytechnic University of Morocco.Data Availability Statement: The data presented in this study are available on request from the corresponding authors.Exosomes are lipid bilayer particles released from cells into their surrounding environment. These vesicles are mediators of near and long-distance intercellular communication and affect various aspects of cell biology. In addition to their biological function, they play an increasingly important role both in diagnosis and as therapeutic agents. In this paper, we review recent literature related to the molecular composition of exosomes, paying special attention to their role in pathogenesis, along with their application as biomarkers and as therapeutic tools. In this context, we analyze the potential use of exosomes in biomedicine, as well as the limitations that preclude their wider application. Simple Summary: The aim of this review is to provide an overview of the current scientific evidence concerning the role played by exosomes in the pathogenesis, diagnosis and treatment of diseases. The potential use of exosomes as delivery vectors for small-molecule therapeutic agents will be discussed. In addition, a special emphasis will be placed on the involvement of exosomes in oncological diseases, as well as to their potential therapeutic application as liquid biopsy tools mainly in cancer diagnosis. A better understanding of exosome biology could improve the results of clinical interventions using exosomes as therapeutic agents.Spanish ISCIII Health Research FundEuropean Union (EU) PI12/01097 PI15/02015 PI18/00337 PIE16-00045 DTS19/00145 PI18/00330Ministry of Science, Innovation and Universities (MICIU) RTI2018-101309-B-C22Junta de Andalucia SOMM17/6109/UGRChair of "Doctors Galera-Requena in cancer stem cell research" CMC-CTS963Junta de Andalucia CARTPI-0001-201CECEyU of the Junta de Andalucia FEDER/European Cohesion Fund (FSE) 2016000073391-TRA 2016000073332-TRA PI57069 PAIDI-Bio326 PI-0014-2016CSyF of the Junta de Andalucia FEDER/European Cohesion Fund (FSE) 2016000073391-TRA 2016000073332-TRA PI57069 PAIDI-Bio326 PI-0014-2016Nicolas Monardes Regional Ministry of Health 0006/2018National Center for Scientific and Technical Research (CNRST) 2UAE2020Mohammed VI Polytechnic University of Morocc

Exosome: A New Player in Translational Nanomedicine

Exosomes are extracellular vesicles released by the vast majority of cell types both in vivo and ex vivo, upon the fusion of multivesicular bodies (MVBs) with the cellular plasma membrane. Two main functions have been attributed to exosomes: their capacity to transport proteins, lipids and nucleic acids between cells and organs, as well as their potential to act as natural intercellular communicators in normal biological processes and in pathologies. From a clinical perspective, the majority of applications use exosomes as biomarkers of disease. A new approach uses exosomes as biologically active carriers to provide a platform for the enhanced delivery of cargo in vivo. One of the major limitations in developing exosome-based therapies is the difficulty of producing sufficient amounts of safe and efficient exosomes. The identification of potential proteins involved in exosome biogenesis is expected to directly cause a deliberate increase in exosome production. In this review, we summarize the current state of knowledge regarding exosomes, with particular emphasis on their structural features, biosynthesis pathways, production techniques and potential clinical applications.This research was funded by the Spanish ISCIII Health Research Fund and the European Regional Development Fund (FEDER) through research grants PI12/01097, PI15/02015, PI18/00337 (F.M.), PIE16-00045 (J.A.M.), DTS19/00145 (J.A.M.) and PI18/00330 (K.B.), as well as by the Spanish Ministry of Science, Innovation and Universities (MICIU) through FEDER research grant RTI2018-101309-B-C2 (J.A.M.). The CECEyU and CSyF Councils of the Junta de Andalucía FEDER/European Cohesion Fund (FSE) provided the following research grants: 2016000073391-TRA, 2016000073332-TRA, PI-57069 and PAIDI-Bio326 (F.M.) and PI-0014-2016 (K.B). K.B. was also on a Nicolas Monardes Regional Ministry of Health contract (0006/2018). H.A. is supported by Research Excellence PhD Fellowship (2UAE2020) from the National Center for Scientific and Technical Research (CNRST). M.T.-M. is funded by the Spanish Ministry of Science and Innovation (SMSI) through an FPU16/05467 fellowship. M.C.-G. is funded by SMSI through a GJ fellowship (PEJ-2018-001760-A).Ye