Choline kinase inhibitors EB-3D and EB-3P interferes with lipid homeostasis in HepG2 cells

A full understanding of the molecular mechanism of action of choline kinase α (ChoKα) inhibitors at the cell level is essential for developing therapeutic and preventive approaches for cancer. The aim of the present study was to evaluate the effects of the ChoKα inhibitors EB-3D and EB-3P on lipid metabolism in HepG2 cells. We used [methyl-14C]choline, [1,2-14C]acetic acid and [2-3H]glycerol as exogenous precursors of the corresponding phospholipids and neutral lipids. [Methyl-14C]choline was also used to determine choline uptake. Protein levels were determined by Western blot. Ultrastructural alterations were investigated by transmission electron microscopy. In this work, we demonstrate that EB-3D and EB-3P interfere with phosphatidylcholine biosynthesis via both CDP-choline pathway and choline uptake by the cell. Moreover, the synthesis of both diacylglycerols and triacylglycerols was affected by cell exposure to both inhibitors. These effects were accompanied by a substantial decrease in cholesterol biosynthesis, as well as alterations in the expression of proteins related to cholesterol homeostasis. We also found that EB-3D and EB-3P lowered ChoKα protein levels. All these effects could be explained by the modulation of the AMP-activated protein kinase signalling pathway. We show that both inhibitors cause mitochondrial alteration and an endoplasmic reticulum stress response. EB-3D and EB-3P exert effects on ChoKα expression, AMPK activation, apoptosis, endoplasmic reticulum stress and lipid metabolism. Taken together, results show that EB-3D and EB-3P have potential anticancer activity through the deregulation of lipid metabolism.This work was aided by the Andalusian Regional Government (P11-CVI-7859). The inhibitors were developed under the Cei-Biotic Project CEI2013-MP-1 (University of Granada)

Design, synthesis, crystallization and biological evaluation of new symmetrical biscationic compounds as selective inhibitors of human Choline Kinase α1 (ChoKα1)

The authors gratefully acknowledge the ‘Consejería de Innovación, Ciencia y Empresa, Junta de Andalucía’ (Excellence Research Project: P07-CTS-03210), the ‘Diputación General de Aragón (B89)’ and the ‘Ministerio de Ciencia e Innovación’ (SAF2009-11955, BFU2010-19504 and CTQ2013-44367-C2-2-P) for the financial support, the award of grants from ‘Ministerio de Educación’ to P.R.-M. and S.S.-E. is gratefully acknowledged, and the ‘Centro de Servicios de Informática of the University of Granada (Spain) for the use of their computers and scientific software. G.V.,R.B., R.M. and G.B. We thanks also the Fondazione Cariparo by the “Progetto Ricerca Pediatrica”. We thank synchrotron radiation sources ALBA (Barcelona), and in particular the beamline XALOC. The research leading to these results has also received funding from the FP7 (2007–2013) under BIOSTRUCTX-7687.A novel family of compounds derivative of 1,1′-(((ethane-1,2-diylbis(oxy))bis(4,1-phenylene))bis(methylene))-bispyridinium or –bisquinolinium bromide (10a-l) containing a pair of oxygen atoms in the spacer of the linker between the biscationic moieties, were synthesized and evaluated as inhibitors of choline kinase against a panel of cancer-cell lines. The most promising compounds in this series were 1,1′-(((ethane-1,2-diylbis(oxy))bis(4,1-phenylene))bis(methylene))bis(4-(dimethylamino)pyridinium) bromide (10a) and 1,1′-(((ethane-1,2-diylbis(oxy))bis(4,1-phenylene))bis(methylene))-bis(7-chloro-4-(pyrrolidin-1-yl)quinolinium) bromide (10l), which inhibit human choline kinase (ChoKα1) with IC50 of 1.0 and 0.92 μM, respectively, in a range similar to that of the previously reported biscationic compounds MN58b and RSM932A. Our compounds show greater antiproliferative activities than do the reference compounds, with unprecedented values of GI50 in the nanomolar range for several of the cancer-cell lines assayed, and more importantly they present low toxicity in non-tumoral cell lines, suggesting a cancer-cell-selective antiproliferative activity. Docking studies predict that the compounds interact with the choline-binding site in agreement with the binding mode of most previously reported biscationic compounds. Moreover, the crystal structure of ChoKα1 with compound 10a reveals that this compound binds to the choline-binding site and mimics HC-3 binding mode as never before.Junta de Andalucía P07-CTS-03210Diputación General de Aragón (B89)Ministerio de Ciencia e Innovación (SAF2009-11955, BFU2010-19504, CTQ2013-44367-C2-2-P)Ministerio de EducaciónUniversity of Granada (Spain)Fondazione CariparoBIOSTRUCTX-7687 FP7 (2007–2013

Estudio y relaciones de la heterocromática asociada al nucleodo en nucleolo en neuronas de ratón

Univ. de Granada, Facultad de Ciencias. Leída el 3-4-197

Anticancer Activity of the Choline Kinase Inhibitor PL48 Is Due to Selective Disruption of Choline Metabolism and Transport Systems in Cancer Cell Lines

This research work was supported by Ministerio de Ciencia e Innovacion (PID2019-109294RB-100) Project. This research was also aided by the Andalusian regional government (CTS-236), B-CTS-216-UGR20 project and by the University of Perugia. Alberto Sola-Leyva holds a Formacion de Doctores 2018 grant (ref. PRE2018-085440) from the Ministerio de Ciencia, Innovacion y Universidades (Spain). Pilar M. Luque-Navarro holds a grant from Ministero dellIstruzione (Italy). Emilio Parisini acknowledges the European Regional Development Fund (ERDF) project BioDrug (No. 1.1.1.5/19/A/004) and the Latvian Council of Science (grant No. lzp-2020/2-0013) for financial support.A large number of different types of cancer have been shown to be associated with an abnormal metabolism of phosphatidylcholine (PC), the main component of eukaryotic cell membranes. Indeed, the overexpression of choline kinase alpha 1 (ChoK alpha 1), the enzyme that catalyses the bioconversion of choline to phosphocholine (PCho), has been found to associate with cell proliferation, oncogenic transformation and carcinogenesis. Hence, ChoK alpha 1 has been described as a possible cancer therapeutic target. Moreover, the choline transporter CTL1 has been shown to be highly expressed in several tumour cell lines. In the present work, we evaluate the antiproliferative effect of PL48, a rationally designed inhibitor of ChoK alpha 1, in MCF7 and HepG2 cell lines. In addition, we illustrate that the predominant mechanism of cellular choline uptake in these cells is mediated by the CTL1 choline transporter. A possible correlation between the inhibition of both choline uptake and ChoK alpha 1 activity and cell proliferation in cancer cell lines is also highlighted. We conclude that the efficacy of this inhibitor on cell proliferation in both cell lines is closely correlated with its capability to block choline uptake and ChoK alpha 1 activity, making both proteins potential targets in cancer therapy.Instituto de Salud Carlos IIISpanish GovernmentEuropean Commission PID2019-109294RB-100Andalusian regional government CTS-236 B-CTS-216-UGR20Spanish Government PRE2018-085440European Regional Development Fund (ERDF) project BioDrug 1.1.1.5/19/A/004Latvian Ministry of Education and Science lzp-2020/2-001

Inhibidores polares simétricos de colina cinassa con actividad antitumoral

Número de publicación: ES2482290 B1. Número de solicitud: 201400466.La invención se refiere a compuestos polares simétricas de bispyridinium, bisquinuclidinium y bisquinolinium con un enlazador derivado de 1,2-bis (p-toliloxi) etano, que son inhibidores de colina quinasa. La invención también se refiere a la síntesis de los mismos y al uso de los mismos como medicamentos antitumorales.Universidad de Granad

New more polar symmetrical bipyridinic compounds: new strategy for the inhibition of choline kinase \u3b11

Aim: Research of the antitumor properties of biscationic compounds has received significant attention over the last few years. Results: A novel family of 1,1′-([2,2′-bipyridine]-5,5′-diylbis(methylene))bis-substituted bromide (9a-k), containing two nitrogen atoms in the linker, considered as hypothetical hydrogen bond acceptors, were synthesized and evaluated as ChoK inhibitors and their antiproliferative activity against six cancer cell lines. Conclusion: The most promising compounds in this series are 1,1′-([2,2′-bipyridine]-5,5′-diylbis(methylene))bis(4-(methyl(phenyl)amino)-quinolinium bromide derivatives 9g-i (analogs to RSM932A), that significantly inhibit cancer cell growth at even submicromolar concentrations, especially against leukemia cells. Compounds 9g-i also inhibit the ChoKα1 with good or moderate values, as predicted by initial docking studies. In addition, the most active compound 9h remarkably induces apoptosis in two cell lines following the mitochondrial pathway. </jats:p

BIOrienta2: orientación a futuros farmacéuticos. La Investigación en biomedicina como carrera profesional

El Grado de Farmacia ofrece una formación idónea para personas que enfoquen su carrera profesional como investigadores en el área de Biomedicina. Ninguna otra formación de Pregrado aúna conocimientos tan amplios en Microbiología, Parasitología, Química, Biología Molecular y Biotecnología así como en aspectos relacionados con el mecanismo de acción de fármacos y con alteracions fisio/patológicas a nivel molecular. Por ello, nos hemos planteado realizar un Plan de Acción Tutorial enfocado a formar alumnos desde sus etapas iniciales en la investigación en Biomedicina: elección de su trayectoria curricular, requisitos para realizar una carrera científica y oportunidades de formación en esta áre