Generación, análisis y visualización de estructuras tridimensionales de complejos fármaco-diana para la docencia teórica y práctica del diseño de fármacos

Memoria ID-127. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2019-2020

JADOPPT: java based AutoDock preparing and processing tool

[EN]Motivation: AutoDock is a very popular software package for docking and virtual screening. However, currently it is hard work to visualize more than one result from the virtual screening at a time. To overcome this limitation we have designed JADOPPT, a tool for automatically preparing and processing multiple ligand-protein docked poses obtained from AutoDock. It allows the simultaneous visual assessment and comparison of multiple poses through clustering methods. Moreover, it permits the representation of reference ligands with known binding modes, binding site residues, highly scoring regions for the ligand, and the calculated binding energy of the best ranked results. Availability and Implementation: JADOPPT, supplementary material (Case Studies 1 and 2) and video tutorials are available at http://visualanalytics.land/cgarcia/JADOPPT.html Contacts: [email protected] or [email protected] Supplementary information: Supplementary data are available at Bioinformatics online

Substitution at the indole 3 position yields highly potent indolecombretastatins with reduced susceptibility to MDR resistance

[EN]Resistance to combretastatin A-4 is mediated by metabolic modification of the phenolic hydroxyl and ether groups of the 3-hydroxy-4-methoxyphenyl (B ring). Replacement of the B ring of combretastatin A-4 by a N-methyl-5-indolyl reduces tubulin polymerization inhibition (TPI) and cytotoxicity against human cancer cell lines but cyano, methoxycarbonyl, formyl, and hydroxyiminomethyl substitutions at the indole 3-position restores potent TPI and cytotoxicity against sensitive human cancer cell lines. These highly potent substituted derivatives displayed low nanomolar cytotoxicity against several human cancer cell lines due to tubulin inhibition, as shown by cell cycle analysis, confocal microscopy, and tubulin polymerization inhibitory activity studies and promoted cell killing mediated by caspase-3 activation. Binding at the colchicine site was suggested by molecular modeling studies. Substituted combretastatins displayed higher potencies than the isomeric isocombretastatins and the highest potencies were achieved for the hydroxyiminomethyl (21) and cyano (23) groups, with TPI values in the submicromolar range and cytotoxicities in the subnanomolar range. Dose-response and time-course studies showed that drug concentrations as low as 1 nM (23) or 10 nM (21) led to a complete G2/M cell cycle arrest after 15 h treatment followed by a high apoptosis-like cell § These authors contributed equally to this work. 3 response after 48-72 h treatment. The P-glycoprotein and calcium antagonist verapamil increased 21 and 23 cytotoxicity to IC50 values of 10-10 M, and highly potentiated the cytotoxic activity in 100-fold of the CHO derivative (17), in A549 human non-small cell lung cancer cells. The differences in cytotoxic potency observed between the highly potent cyano (23) and hydroxyiminomethyl (21) groups and other substituents with similar TPI values (17) were very much reduced upon co-treatment with verapamil. A 3,4,5-trimethoxyphenyl ring always afforded more potent derivatives than a 2,3,4-trimethoxyphenyl ring

Seguridad en el laboratorio. Revisión de la enseñanza práctica de la Química Orgánica en la Facultad de Farmacia

Memoria ID-136. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2018-2019

Bioactive Heterometallic CuII–ZnII Complexes with Potential Biomedical Applications

[EN]A series of multinuclear heterometallic Cu-Zn complexes of molecular formula [(CuL)2Zn(dca)2] (1), [(CuL)2Zn(NO3)2] (2), [(CuL)2Zn2(Cl)4] (3), and [(CuL)2Zn2(NO2)4] (4) have been synthesized by reacting [CuL] as a "metalloligand (ML)" (where HL = N,N′-bis(5-chloro-2-hydroxybenzylidene)-2,2-dimethylpropane-1,3-diamine) and by varying the anions or coligands using the same molar ratios of the reactants. All of the four products including the ML have been characterized by infrared and UV-vis spectroscopies and elemental and single-crystal X-ray diffraction analyses. By varying the anions, different structures and topologies are obtained which we have tried to rationalize by means of thorough density functional theory calculations. All of the complexes (1-4) have now been applied for several biological investigations to verify their therapeutic worth. First, their cytotoxicity properties were assessed against HeLa human cervical carcinoma along with the determination of IC50 values. The study was extended with extensive DNA and protein binding experiments followed by detailed fluorescence quenching study with suitable reagents to comprehend the mechanistic pathway. From all of these biological studies, it has been found that all of these heterometallic complexes show more than a few fold improvement of their therapeutic values as compared to the similar homometallic ones probably because of the simultaneous synergic effect of copper and zinc. Among all of the four heterometallic complexes, complex 3 exhibits highest binding constants and IC50 values suggest for their better interaction toward the biological targets and hence have better clinical importance

The Masked Polar Group Incorporation (MPGI) Strategy in Drug Design: Effects of Nitrogen Substitutions on Combretastatin and Isocombretastatin Tubulin Inhibitors

[EN] Colchicine site ligands suffer from low aqueous solubility due to the highly hydrophobic nature of the binding site. A new strategy for increasing molecular polarity without exposing polar groups—termed masked polar group incorporation (MPGI)—was devised and applied to nitrogenated combretastatin analogues. Bulky ortho substituents to the pyridine nitrogen hinder it from the hydrophobic pocket while increasing molecular polarity. The resulting analogues show improved aqueous solubilities and highly potent antiproliferative activity against several cancer cell lines of different origin. The more potent compounds showed moderate tubulin polymerization inhibitory activity, arrested the cell cycle of treated cells at the G2/M phase, and subsequently caused apoptotic cell death represented by the cells gathered at the subG0/G1 population after 48 h of treatment. Annexin V/Propidium Iodide (PI) double-positive cells observed after 72 h confirmed the induction of apoptosis. Docking studies suggest binding at the colchicine site of tubulin in a similar way as combretastatin A4, with the polar groups masked by the vicinal substituents. These results validate the proposed strategy for the design of colchicine site ligands and open a new road to increasing the aqueous solubility of ligands binding in apolar environments

Adaptación de las clases prácticas de asignaturas de química a un modelo de semipresencialidad y optimización del tiempo de trabajo en laboratorio

Memoria ID-145 Ayudas de la Universidad de Salamanca para la innovación docente, curso 2020-2021

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El objetivo general de la asignatura es completar los conocimientos generales de Química Orgánica con los referentes a determinación estructural de los compuestos orgánicos y la química de los compuestos heterocíclicos. En primer lugar se estudiará la determinación estructural basada fundamentalmente en la aplicación de técnicas espectroscópicas. Posteriormente se realizará el estudio de los compuestos heterocíclicos aromáticos agrupados por su tamaño y número de heteroátomos. Este grupo de compuestos orgánicos forma parte de la estructura de muchos fármacos y moléculas de importancia biológica, por lo que su estudio servirá para completar la formación en Química Orgánica con una familia de compuestos de interés. Asignatura impartida en el Segundo curso de la Licenciatura en Farmacia.I. Materiales de clase: Tema 1.Determinación de la fórmula molecular y de los grupos funcionales.; Tema 2.Resonancia magnética nuclear RMN-1H y 13C; Tema 3.Aplicación de los métodos físicos en la determinación de la estructura;Tema 4.Clasificación y nomenclatura de los compuestos heterocíclicos; Tema 5. Estructura y propiedades generales; Tema 6. Síntesis de heterociclos; Tema 7. Heterociclos pentagonales con un sólo heteroátomo; Tema 8. Benzofuranos, benzotiofenos e indoles; Tema 9. Sistemas pentagonales con varios heteroátomos; Tema 10. Sistemas hexagonales nitrogenados; Tema 11. Quinolinas e Isoquinolinas; Tema 12. Sistemas hexagonales oxigenados y sus benzoderivados; Tema 13. Heterociclos de seis miembros con dos heteroátomos; Tema 14. Derivados de importancia biológica de la pirimidina. II. Bibliografí

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Asignatura optativa impartida en el Cuarto curso del Grado en Farmacia de la Universidad de Salamanca