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1.
Bioorg Med Chem Lett ; 111: 129894, 2024 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-39043264

RESUMEN

Drug repurposing and rescuing have been widely explored as cost-effective approaches to expand the portfolio of chemotherapeutic agents. Based on the reported antitumor properties of both trans-cinnamic acids and quinacrine, an antimalarial aminoacridine, we explored the antiproliferative properties of two series of N-cinnamoyl-aminoacridines recently identified as multi-stage antiplasmodial leads. The compounds were evaluated in vitro against three cancer cell lines (MKN-28, Huh-7, and HepG2), and human primary dermal fibroblasts. One of the series displayed highly selective antiproliferative activity in the micromolar range against the three cancer cell lines tested, without any toxicity to non-carcinogenic cells.


Asunto(s)
Antimaláricos , Antineoplásicos , Proliferación Celular , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Proliferación Celular/efectos de los fármacos , Antineoplásicos/farmacología , Antineoplásicos/química , Antineoplásicos/síntesis química , Relación Estructura-Actividad , Antimaláricos/farmacología , Antimaláricos/química , Antimaláricos/síntesis química , Línea Celular Tumoral , Reposicionamiento de Medicamentos , Estructura Molecular , Aminoacridinas/farmacología , Aminoacridinas/química , Aminoacridinas/síntesis química , Relación Dosis-Respuesta a Droga , Cinamatos/farmacología , Cinamatos/química , Cinamatos/síntesis química
2.
Int J Mol Sci ; 22(24)2021 Dec 07.
Artículo en Inglés | MEDLINE | ID: mdl-34947987

RESUMEN

Acridine cell-penetrating peptide conjugates are an extremely important family of compounds in antitumor chemotherapy. These conjugates are not so widely analysed in antimicrobial therapy, although bioactive peptides could be used as nanocarriers to smuggle antimicrobial compounds. An octaarginine conjugate of an imidazoacridinone derivative (Compound 1-R8) synthetized by us exhibited high antifungal activity against reference and fluconazole-resistant clinical strains (MICs ≤ 4 µg mL-1). Our results clearly demonstrate the qualitative difference in accumulation of the mother compound and Compound 1-R8 conjugate into fungal cells. Only the latter was transported and accumulated effectively. Microscopic and flow cytometry analysis provide some evidence that the killing activity of Compound 1-R8 may be associated with a change in the permeability of the fungal cell membrane. The conjugate exhibited low cytotoxicity against human embryonic kidney (HEK-293) and human liver (HEPG2) cancer cell lines. Nevertheless, the selectivity index value of the conjugate for human pathogenic strains remained favourable and no hemolytic activity was observed. The inhibitory effect of the analysed compound on yeast topoisomerase II activity suggested its molecular target. In summary, conjugation with R8 effectively increased imidazoacridinone derivative ability to enter the fungal cell and achieve a concentration inside the cell that resulted in a high antifungal effect.


Asunto(s)
Aminoacridinas/síntesis química , Antifúngicos/síntesis química , Candida albicans/crecimiento & desarrollo , Péptidos de Penetración Celular/síntesis química , Oligopéptidos/química , Aminoacridinas/química , Aminoacridinas/farmacología , Antifúngicos/química , Antifúngicos/farmacología , Candida albicans/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Péptidos de Penetración Celular/química , Péptidos de Penetración Celular/farmacología , Células HEK293 , Células Hep G2 , Humanos , Viabilidad Microbiana/efectos de los fármacos , Estructura Molecular
3.
Bioorg Med Chem ; 40: 116191, 2021 06 15.
Artículo en Inglés | MEDLINE | ID: mdl-33965841

RESUMEN

Cisplatin analogues with an attached DNA-binding moiety represent a potentially effective class of DNA-damaging anti-tumour agents because they possess higher affinities for DNA and different DNA damage profiles compared with cisplatin. In this study, the interaction of four 9-aminoacridine carboxamide Pt complexes with purified DNA was investigated: firstly, using a fluorescent intercalator displacement (FID) assay with ethidium bromide; and secondly, with a DNA unwinding assay. The relative capacity of these compounds to perturb the fluorescence induced by DNA-bound ethidium bromide at clinically relevant drug concentrations was assessed over a 24-h period using an FID assay. All analogues were found to reduce the level of ethidium bromide-induced fluorescence in a concentration-dependent manner from the earliest time point of 10 min onwards. Cisplatin, however, showed a markedly slower reduction in ethidium bromide-induced fluorescence from 2 h onwards, producing a similar level of fluorescence reduction as that produced by the analogues from 6 h onwards. These results suggest that the altered DNA-binding modes of the DNA-targeted analogues confer a more efficient mechanism for DNA binding compared with cisplatin. Relative DNA binding coefficients were also determined for each of the compounds studied. With the DNA unwinding assay, an unwinding angle can be calculated from the coalescence point of plasmids in an agarose gel. It was found that all 9-aminoacridine carboxamide analogues had a greater unwinding angle compared with cisplatin. The knowledge obtained from these two assays has helped to further characterise the cisplatin analogues and could facilitate the development of more effective anti-tumour agents.


Asunto(s)
Aminoacridinas/farmacología , Antineoplásicos/farmacología , ADN/efectos de los fármacos , Compuestos Organoplatinos/farmacología , Aminoacridinas/química , Antineoplásicos/química , Sitios de Unión/efectos de los fármacos , ADN/química , Relación Dosis-Respuesta a Droga , Estructura Molecular , Compuestos Organoplatinos/química , Plásmidos , Relación Estructura-Actividad
4.
ChemMedChem ; 16(5): 788-792, 2021 03 03.
Artículo en Inglés | MEDLINE | ID: mdl-33217195

RESUMEN

Multi-stage drugs have been prioritized in antimalarial drug discovery, as targeting more than one process in the Plasmodium life cycle is likely to increase efficiency, while decreasing the chances of emergence of resistance by the parasite. Herein, we disclose two novel acridine-based families of compounds that combine the structural features of primaquine and chloroquine. Compounds prepared and studied thus far retained the in vitro activity displayed by the parent drugs against the erythrocytic stages of chloroquine-sensitive and -resistant Plasmodium falciparum strains, and against the hepatic stages of Plasmodium berghei, hence acting as dual-stage antiplasmodial hits.


Asunto(s)
Aminoacridinas/farmacología , Antimaláricos/farmacología , Plasmodium berghei/efectos de los fármacos , Plasmodium falciparum/efectos de los fármacos , Aminoacridinas/química , Antimaláricos/química , Relación Dosis-Respuesta a Droga , Estructura Molecular , Pruebas de Sensibilidad Parasitaria , Relación Estructura-Actividad
5.
Biophys J ; 117(6): 1101-1115, 2019 09 17.
Artículo en Inglés | MEDLINE | ID: mdl-31474304

RESUMEN

Understanding local conformations of DNA at the level of individual nucleic acid bases and base pairs is important for elucidating molecular processes that depend on DNA sequence. Here, we apply linear absorption and circular dichroism measurements to the study of local DNA conformations, using the guanine base analog 6-methyl isoxanthopterin (6-MI) as a structural probe. We show that the spectroscopic properties of this probe can provide detailed information about the average local base and basepair conformations as a function of the surrounding DNA sequence. Based on these results we apply a simple theoretical model to calculate the circular dichroism spectra of 6-MI-substituted DNA constructs and show that our model can be used to extract information about how the local conformations of the 6-MI probe are influenced by the local base or basepair environment. We also use this probe to examine the pathway for the insertion (intercalation) of a tethered acridine ligand (9-amino-6-chloro methoxyacridine) into duplex DNA. We show that this model intercalator interacts with duplex DNA by a "displacement insertion intercalation" mechanism, whereby the acridine moiety is inserted into the DNA structure and displaces the base located opposite its attachment site. These findings suggest that site-specifically positioned base analog probes can be used to characterize the molecular and structural details of binding ligand effects on local base stacking and unstacking reactions in single- and double-stranded DNA and thus may help to define the molecular mechanisms of DNA-protein interactions that involve the site-specific intercalation of aromatic amino acid side chains into genomic DNA.


Asunto(s)
ADN/química , Sustancias Intercalantes/química , Sondas Moleculares/química , Conformación de Ácido Nucleico , Aminoacridinas/química , Secuencia de Bases , Simulación por Computador , Electricidad , Ligandos , Modelos Moleculares , Xantopterina/química
6.
Molecules ; 23(3)2018 Mar 11.
Artículo en Inglés | MEDLINE | ID: mdl-29534488

RESUMEN

Symptomatic treatment of myasthenia gravis is based on the use of peripherally-acting acetylcholinesterase (AChE) inhibitors that, in some cases, must be discontinued due to the occurrence of a number of side-effects. Thus, new AChE inhibitors are being developed and investigated for their potential use against this disease. Here, we have explored two alternative approaches to get access to peripherally-acting AChE inhibitors as new agents against myasthenia gravis, by structural modification of the brain permeable anti-Alzheimer AChE inhibitors tacrine, 6-chlorotacrine, and huprine Y. Both quaternization upon methylation of the quinoline nitrogen atom, and tethering of a triazole ring, with, in some cases, the additional incorporation of a polyphenol-like moiety, result in more polar compounds with higher inhibitory activity against human AChE (up to 190-fold) and butyrylcholinesterase (up to 40-fold) than pyridostigmine, the standard drug for symptomatic treatment of myasthenia gravis. The novel compounds are furthermore devoid of brain permeability, thereby emerging as interesting leads against myasthenia gravis.


Asunto(s)
Acetilcolinesterasa/metabolismo , Aminoacridinas/síntesis química , Aminoquinolinas/síntesis química , Inhibidores de la Colinesterasa/síntesis química , Acetilcolinesterasa/química , Aminoacridinas/química , Aminoacridinas/farmacología , Aminoquinolinas/química , Aminoquinolinas/farmacología , Butirilcolinesterasa/química , Butirilcolinesterasa/metabolismo , Inhibidores de la Colinesterasa/química , Inhibidores de la Colinesterasa/farmacología , Regulación hacia Abajo , Proteínas Ligadas a GPI/química , Proteínas Ligadas a GPI/metabolismo , Compuestos Heterocíclicos de 4 o más Anillos/química , Humanos , Modelos Moleculares , Estructura Molecular , Miastenia Gravis/tratamiento farmacológico , Miastenia Gravis/enzimología , Relación Estructura-Actividad , Tacrina/química
7.
Chembiochem ; 18(16): 1599-1603, 2017 08 17.
Artículo en Inglés | MEDLINE | ID: mdl-28681411

RESUMEN

For decades the detection of nucleic acids and their interactions at low abundances has been a challenging task that has thus far been solved by enzymatic target amplification. In this work we aimed at developing efficient tools for amplification-free nucleic acid detection, which resulted in the synthesis of new fluorescent nanoparticles. Here, the fluorescent nanoparticles were made by simple and inexpensive radical emulsion polymerization of butyl acrylate in the presence of fluorescent dyes and additional functionalization reagents. This provided ultra-bright macrofluorophores of 9-84 nm mean diameter, modified with additional alkyne and amino groups for bioconjugation. By using click and NHS chemistries, the new nanoparticles were attached to target-specific DNA probes that were used in fluorimetry and fluorescence microscopy. Overall, these fluorescent nanoparticles and their oligonucleotide derivatives have higher photostability, brighter fluorescence and hence dramatically lower limits of target detection than the individual organic dyes. These properties make them useful in approaches directed towards ultrasensitive detection of nucleic acids, in particular for imaging and in vitro diagnostics of DNA.


Asunto(s)
Sondas de ADN/química , Colorantes Fluorescentes/química , MicroARNs/análisis , Nanopartículas/química , Acrilatos/química , Aminoacridinas/química , Aminoacridinas/efectos de la radiación , Azidas/química , Benzotiazoles/química , Benzotiazoles/efectos de la radiación , Carbocianinas/química , Carbocianinas/efectos de la radiación , Química Clic , Dispersión Dinámica de Luz , Colorantes Fluorescentes/efectos de la radiación , Fluorometría , Límite de Detección , Microscopía Electrónica de Transmisión , Microscopía Fluorescente , Nanopartículas/efectos de la radiación , Tamaño de la Partícula , Perileno/química , Perileno/efectos de la radiación , Polimerizacion , Propilaminas/química , Quinolinas/química , Quinolinas/efectos de la radiación , Rayos Ultravioleta
8.
Anal Biochem ; 530: 17-30, 2017 08 01.
Artículo en Inglés | MEDLINE | ID: mdl-28465034

RESUMEN

Glycosaminoglycans are a heterogeneous family of linear polysaccharides comprised of repeating disaccharide subunits that mediate many effects at the cellular level. There is increasing evidence that the nature of these effects is determined by differences in disaccharide composition. However, the determination of GAG disaccharide composition in biological samples remains challenging and time-consuming. We have developed a method that uses derivatization and selected ion recording and RP-UPLCMS resulting in rapid separation and quantification of twelve heparin/heparin sulfate disaccharides from 5 µg GAG. Limits of detection and quantitation were 0.02-0.15 and 0.07-0.31 µg/ml respectively. We have applied this method to the novel analysis of disaccharide levels extracted from heparan sulfate and human cancer cell lines. Heparan sulfate disaccharides extracted from biological samples following actinase and heparinase incubation and derivatized using reductive amination with 2-aminoacridone. Derivatized disaccharides were analyzed used UPLC-MS with single ion monitoring. Eight HS disaccharide subunits were separated and quantified from HS and cell lines in eleven minutes per sample. In all samples the most abundant subunits present were the unsulfated ΔUA-GlcNAc, ΔUA-GlcNAc,6S and ΔUA,2S-GlcNS,6S. There was considerable variation in the proportions and concentrations of disaccharides between different cell lines. Further studies are needed to examine the significance of these differences.


Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Disacáridos/análisis , Heparina/análogos & derivados , Heparitina Sulfato/análisis , Espectrometría de Masas/métodos , Neoplasias/metabolismo , Aminoacridinas/química , Disacáridos/química , Disacáridos/aislamiento & purificación , Heparina/análisis , Heparina/química , Heparina/aislamiento & purificación , Liasa de Heparina/metabolismo , Heparitina Sulfato/química , Heparitina Sulfato/aislamiento & purificación , Humanos , Células Tumorales Cultivadas
9.
Oncotarget ; 8(19): 31187-31198, 2017 May 09.
Artículo en Inglés | MEDLINE | ID: mdl-28415717

RESUMEN

C-1311 is a small molecule, which has shown promise in a number of pre-clinical and clinical studies. However, the biological response to C-1311 exposure is complicated and has been reported to involve a number of cell fates. Here, we investigated the molecular signaling which determines the response to C-1311 in both cancer and non-cancer cell lines. For the first time we demonstrate that the tumor suppressor, p53 plays a key role in cell fate determination after C-1311 treatment. In the presence of wild-type p53, cells exposed to C-1311 entered senescence. In contrast, cells lines without functional p53 underwent mitotic catastrophe and apoptosis. C-1311 also induced autophagy in a non-p53-dependent manner. Cells in hypoxic conditions also responded to C-1311 in a p53-dependent manner, suggesting that our observations are physiologically relevant. Most importantly, we show that C-1311 can be effectively combined with radiation to improve the radiosensitivity of a panel of cancer cell lines. Together, our data suggest that C-1311 warrants further clinical testing in combination with radiotherapy for the treatment of solid tumors.


Asunto(s)
Aminoacridinas/farmacología , Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Apoptosis/genética , Tolerancia a Radiación/efectos de los fármacos , Tolerancia a Radiación/genética , Fármacos Sensibilizantes a Radiaciones/farmacología , Proteína p53 Supresora de Tumor/genética , Aminoacridinas/química , Antineoplásicos/química , Ciclo Celular/efectos de los fármacos , Ciclo Celular/genética , Línea Celular Tumoral , Senescencia Celular/efectos de los fármacos , Senescencia Celular/genética , Relación Dosis-Respuesta en la Radiación , Técnicas de Inactivación de Genes , Humanos , Mitosis/efectos de los fármacos , Mitosis/genética , Fármacos Sensibilizantes a Radiaciones/química
10.
J Chromatogr A ; 1445: 68-79, 2016 May 06.
Artículo en Inglés | MEDLINE | ID: mdl-27062721

RESUMEN

Oligosaccharide mapping based on enzyme cleavage provides a useful molecular fingerprint of the heparin structure revealing detailed structural information regarding its sequence and the content of part of the ATIII-binding region. This approach is performed by strong-anion exchange (SAX)-HPLC separation which is incompatible with MS requiring purification of oligosaccharides for their conclusive identification. We report a novel oligosaccharide mapping strategy based on the HILIC separation of the main heparin disaccharides/oligosaccharides released by heparinase I, fluorotagged with 2-aminoacridone and on-line detected by a fluorescence detector and characterized by ESI-MS. The application of a polar solvent having a high pH with acetonitrile avoided desulfation enabling a simple and accurate structural oligosaccharide assignment. Oligosaccharide mapping, or merely complete disaccharide composition, may be performed on nanogram-scale by the fluorescence detector vs micrograms useful for classical SAX-HPLC. Additionally, only widely commercially available heparin lyase I is necessary, without the use of expensive heparinases II and III. Contrary to SAX-HPLC, this novel HILIC approach is able to separate and identify the saturated trisulfated disaccharide belonging to the non-reducing end of heparin chains. Finally, the content of 3-O-sulfo groups of the ATIII-binding region is determined.


Asunto(s)
Técnicas de Química Analítica/métodos , Cromatografía Líquida de Alta Presión , Heparina/química , Oligosacáridos/análisis , Espectrometría de Masa por Ionización de Electrospray , Aminoacridinas/química , Disacáridos/química , Fluorescencia , Heparina/análogos & derivados , Heparina/análisis , Liasa de Heparina/metabolismo , Interacciones Hidrofóbicas e Hidrofílicas , Oligosacáridos/química
11.
Klin Lab Diagn ; 61(4): 238-41, 2016 Apr.
Artículo en Ruso | MEDLINE | ID: mdl-30586245

RESUMEN

Nowadays, necessity is shown up concerning modeling of certain ecological processes actually emerging in various biotops and hence step-by-step study of development of biofilms on solid surfaces (glass and plastic). To implement such kind of studies the light and luminescent microscopy are the most available and rather informative tools. The study used strains V.cholerae O1 and O139 serogroups with different genetic characteristics. The biofilms were developed on cover glass and/or plastic plates which were placed in bottles with tap autoclave water in vertical position in special device. The lifetime visualization of stages of development of biofilm was implemented using luminescent microscopy by coloring biofilm with solution of acridine yellow. The biofilms colored with Congo read and fuchsine were analyzed using light microscopy. The light and luminescent microscopy permit investigating biofilms in dynamics at various stages of development, to evaluate condition of its physiological functioning in quantitative and qualitative dimensions and to trace synthesis of exopolysaccharides in different serogroups of vibrios that has significance in prognosis of velocity of occurrence of biofilms on abiotic objects.


Asunto(s)
Biopelículas/crecimiento & desarrollo , Cólera/microbiología , Vibrio cholerae/crecimiento & desarrollo , Aminoacridinas/química , Vidrio , Humanos , Luminiscencia , Serogrupo , Vibrio cholerae/patogenicidad , Vibrio cholerae/ultraestructura , Agua/química
12.
J Biomol Struct Dyn ; 34(3): 653-63, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-26211888

RESUMEN

Imidazoacridinone C-1311 (Symadex®) is a powerful antitumor agent, which successfully made its way through the Phase I clinical trials and has been recommended for Phase II few a years ago. It has been shown experimentally that during the initial stage of its action C-1311 forms a relatively stable intercalation complex with DNA, yet it has shown no base-sequence specificity while binding to DNA. In this paper, the d(CGATCG)2:C-1311 intercalation complex has been studied by means of two-dimensional NMR spectroscopy, yielding a full assignment of the resonance lines observed in (1)H NMR spectra. The observation of the intermolecular NOE contacts between C-1311 and DNA allowed locating the ligand between the guanine and adenine moieties. Formation of a symmetric complex was pointed out on the basis of the lack of a second set of the (1)H resonances. The resulting stereostructure of the complex was then improved by means of molecular dynamics, using the CHARMM force field and GROMACS software. To this end, distance restraints derived from the NOESY cross-peak volumes were applied to the atomistic model of the d(CGATCG)2:C-1311 complex. Obtained results are in full agreement with biochemical data on the mechanism of action of C-1311, in particular with the previously postulated post-intercalation enzymatic activation of the studied drug.


Asunto(s)
Aminoacridinas/química , Antineoplásicos/química , ADN/química , Sustancias Intercalantes/química , Espectroscopía de Resonancia Magnética , Modelos Moleculares , Estructura Molecular , Conformación de Ácido Nucleico , Estereoisomerismo
13.
Methods Mol Biol ; 1377: 171-80, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-26695032

RESUMEN

The activity of enzymes involved in active transport of matter across lipid bilayers can conveniently be assayed by measuring their consumption of energy, such as ATP hydrolysis, while it is more challenging to directly measure their transport activities as the transported substrate is not converted into a product and only moves a few nanometers in space. Here, we describe two methods for the measurement of active proton pumping across lipid bilayers and the concomitant formation of a membrane potential, applying the dyes 9-amino-6-chloro-2-methoxyacridine (ACMA) and oxonol VI. The methods are exemplified by assaying transport of the Arabidopsis thaliana plasma membrane H(+)-ATPase (proton pump), which after heterologous expression in Saccharomyces cerevisiae and subsequent purification has been reconstituted in proteoliposomes.


Asunto(s)
Adenosina Trifosfato/química , Membrana Dobles de Lípidos/química , ATPasas de Translocación de Protón/biosíntesis , Adenosina Trifosfato/biosíntesis , Aminoacridinas/química , Arabidopsis/enzimología , Transporte Biológico Activo , Regulación Enzimológica de la Expresión Génica , Hidrólisis , Isoxazoles/química , Membrana Dobles de Lípidos/metabolismo , Potenciales de la Membrana , ATPasas de Translocación de Protón/química , ATPasas de Translocación de Protón/aislamiento & purificación , Saccharomyces cerevisiae/genética
14.
J Chromatogr A ; 1397: 43-51, 2015 Jun 05.
Artículo en Inglés | MEDLINE | ID: mdl-25913329

RESUMEN

Pneumococcal type-3 polysaccharide (Pn3P) is considered a major target for the development of a human vaccine to protect against Streptococcus pneumoniae infection. Thus, it is critical to develop methods for the preparation and analysis of Pn3P-derived oligosaccharides to better understand its immunological properties. In this paper, we profile oligosaccharides, generated by the free radical depolymerization of Pn3P, using liquid chromatography (LC)-tandem mass spectrometry (MS/MS). Hydrophilic liquid interaction chromatography (HILIC)-mass spectrometry (MS) revealed a series of oligosaccharides with an even- and odd-number of saccharide residues, ranging from monosaccharide, degree of polymerization (dp1) to large oligosaccharides up to dp 20, generated by free radical depolymerization. Isomers of oligosaccharides with an even number of sugar residues were easily separated on a HILIC column, and their sequences could be distinguished by comparing MS/MS of these oligosaccharides and their reduced alditols. Fluorescent labeling with 2-aminoacridone (AMAC) followed by reversed phase (RP)-LC-MS/MS was applied to analyze and sequence poorly separated product mixtures, as RP-LC affords higher resolution of AMAC-labeled oligosaccharides than does HILIC-based separation. The present methodology can be potentially applied to profiling other capsular polysaccharides.


Asunto(s)
Técnicas de Química Analítica , Cromatografía Liquida , Oligosacáridos/química , Streptococcus pneumoniae/química , Espectrometría de Masas en Tándem , Aminoacridinas/química , Interacciones Hidrofóbicas e Hidrofílicas , Isomerismo
15.
Methods Mol Biol ; 1229: 143-59, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-25325951

RESUMEN

Hyaluronan (HA), chondroitin sulfate (CS), and heparan sulfate (HS) are glycosaminoglycans (GAGs) with a great importance in biological processes as they participate in functional cell properties, such as migration, adhesion, and proliferation. A perturbation of the quantity and/or the sulfation of GAGs is often associated with pathological conditions. In this chapter, we present valuable and validated protocols for the analysis of HA-, CS-, and HS-derived disaccharides after derivatization with 2-aminoacridone and by using the fluorophore-assisted carbohydrate electrophoresis (FACE). FACE is a well-known technique and a reliable tool for a fast screening of GAGs, as it is possible to analyze 16 samples at the same time with one electrophoretic apparatus. The protocols for the gel preparation are based on the variations of the acrylamide/bisacrylamide and buffer concentrations. Different approaches for the extraction and purification of the disaccharides of various biologic samples and pharmaceutical preparations are also stressed.


Asunto(s)
Disacáridos/sangre , Disacáridos/orina , Electroforesis/métodos , Colorantes Fluorescentes/química , Glicosaminoglicanos/sangre , Glicosaminoglicanos/orina , Preparaciones Farmacéuticas/química , Aminoacridinas/química , Animales , Tampones (Química) , Química Farmacéutica , Sulfatos de Condroitina/sangre , Sulfatos de Condroitina/orina , Cromatografía Líquida de Alta Presión , Disacáridos/análisis , Electroforesis en Gel de Poliacrilamida , Glicosaminoglicanos/análisis , Glicosaminoglicanos/aislamiento & purificación , Heparina/aislamiento & purificación , Heparitina Sulfato/aislamiento & purificación , Humanos , Ácido Hialurónico/análisis , Cápsula del Cristalino/metabolismo , Ratones , Ratas
16.
Biochem J ; 463(2): 225-37, 2014 Oct 15.
Artículo en Inglés | MEDLINE | ID: mdl-25072268

RESUMEN

Hydroxyl radicals (•OH) cause non-enzymic scission of polysaccharides in diverse biological systems. Such reactions can be detrimental (e.g. causing rheumatic and arthritic diseases in mammals) or beneficial (e.g. promoting the softening of ripening fruit, and biomass saccharification). Here we present a method for documenting •OH action, based on fluorescent labelling of the oxo groups that are introduced as glycosulose residues when •OH attacks polysaccharides. The method was tested on several polysaccharides, especially pectin, after treatment with Fenton reagents. 2-Aminoacridone plus cyanoborohydride reductively aminated the oxo groups in treated polysaccharides; the product was then reacted with acetone plus cyanoborohydride, forming a stable tertiary amine with the carbohydrate linked to N-isopropyl-2-aminoacridone (pAMAC). Digestion of labelled pectin with 'Driselase' yielded several fluorescent products which on electrophoresis and HPLC provided a useful 'fingerprint' indicating •OH attack. The most diagnostic product was a disaccharide conjugate of the type pAMAC·UA-GalA (UA=unspecified uronic acid), whose UA-GalA bond was Driselase-resistant (product 2A). 2A was clearly distinguishable from GalA-GalA-pAMAC (disaccharide labelled at its reducing end), which was digestible to GalA-pAMAC. The methodology is applicable, with appropriate enzymes in place of Driselase, for detecting natural and artificial •OH attack in diverse plant, animal and microbial polysaccharides.


Asunto(s)
Aminoacridinas/química , Colorantes Fluorescentes/química , Radical Hidroxilo/química , Polisacáridos/química , Coloración y Etiquetado/métodos , Estructura Molecular
17.
Bioorg Med Chem Lett ; 24(14): 3014-7, 2014 Jul 15.
Artículo en Inglés | MEDLINE | ID: mdl-24908610

RESUMEN

A series of 9-alkylaminoacridines were synthesized and evaluated for activity against two strains of methicillin-resistant and one strain of methicillin-sensitive Staphylococcus aureus. Results are presented that show a clear structure activity relationship between the N-alkyl chain length and antibacterial activity with peak MIC99 values of 2-3 µM for alkyl chains ranging from 10 to 14 carbons in length. Although prior work has linked the function of acridine-based compounds to intercalation and topoisomerase inhibition, the present results show that 9-alkylaminoacridines likely function as amphiphilic membrane-active disruptors potentially in a similar manner as quaternary ammonium antimicrobials.


Asunto(s)
Aminoacridinas/síntesis química , Aminoacridinas/farmacología , Antibacterianos/síntesis química , Antibacterianos/farmacología , Aminoacridinas/química , Antibacterianos/química , Relación Dosis-Respuesta a Droga , Meticilina/farmacología , Pruebas de Sensibilidad Microbiana , Estructura Molecular , Staphylococcus aureus/efectos de los fármacos , Relación Estructura-Actividad
18.
Anal Bioanal Chem ; 406(19): 4617-26, 2014 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-24817364

RESUMEN

A capillary zone electrophoresis-laser-induced fluorescence detection (CZE-LIF) method was developed for the simultaneous analysis of disaccharides derived from heparan sulfate, chondroitin sulfate/dermatan sulfate, hyaluronan, and keratan sulfate. Glycosaminoglycans (GAGs) were first depolymerized with the mixture of GAG lyases (heparinase I, II, III and chondroitinase ABC and chondroitinase AC II) and GAG endohydrolase (keratinase II) and the resulting disaccharides were derivatized by reductive amination with 2-aminoacridone. Nineteen fluorescently labeled disaccharides were separated using 50 mM phosphate buffer (pH 3.3) under reversed polarity at 25 kV. Using these conditions, all the disaccharides examined were baseline separated in less then 25 min. This CZE-LIF method gave good reproducibility for both migration time (≤1.03% for intraday and ≤4.4% for interday) and the peak area values (≤5.6% for intra- and ≤8.69% for interday). This CZE-LIF method was used for profiling and quantification of GAG derivative disaccharides in bovine cornea. The results show that the current CZE-LIF method offers fast, simple, sensitive, reproducible determination of disaccharides derived from total GAGs in a single run.


Asunto(s)
Aminoacridinas/química , Córnea/química , Electroforesis Capilar/métodos , Glicosaminoglicanos/análisis , Espectrometría de Masa por Ionización de Electrospray/métodos , Glicosaminoglicanos/química
19.
J Biol Inorg Chem ; 19(6): 997-1007, 2014 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-24827388

RESUMEN

In this study, the DNA sequence specificity of four DNA-targeted 9-aminoacridine carboxamide Pt complexes was compared with cisplatin, using two specially constructed plasmid templates. One plasmid contained 5'-CG and 5'-GA insert sequences while the other plasmid contained a G-rich transferrin receptor gene promoter insert sequence. The damage profiles of each compound on the different DNA templates were quantified via a polymerase stop assay with fluorescently labelled primers and capillary electrophoresis. With the plasmid that contained 5'-CG and 5'-GA dinucleotides, the four 9-aminoacridine carboxamide Pt complexes produced distinctly different damage profiles as compared with cisplatin. These 9-aminoacridine complexes had greatly increased levels of DNA damage at CG and GA dinucleotides as compared with cisplatin. It was shown that the presence of a CG or GA dinucleotide was sufficient to reveal the altered DNA sequence selectivity of the 9-aminoacridine carboxamide Pt analogues. The DNA sequence specificity of the Pt complexes was also found to be similarly altered utilising the transferrin receptor DNA sequence.


Asunto(s)
Aminoacridinas/química , ADN/efectos de los fármacos , ADN/genética , Compuestos Organoplatinos/farmacología , Secuencia de Bases , Cisplatino/farmacología , Daño del ADN , Estructura Molecular , Compuestos Organoplatinos/química , Especificidad por Sustrato
20.
Luminescence ; 29(8): 1053-8, 2014 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-24753178

RESUMEN

Based on the oxidation of acridine yellow by permanganate in basic medium, a new chemiluminescence system was developed for the sensitive determination of some important drugs. The remarkable inhibiting effect of fluvoxamine, ceftriaxone and isoniazid on this reaction was applied to their detection. A possible mechanism was proposed for this system based on chemiluminescence emission wavelengths and experimental observations. Under optimum conditions, calibration graphs were obtained for 1 × 10(-9) to 1 × 10(-6) mol/L of fluvoxamine; 2 × 10(-8) to 8 × 10(-6) mol/L of ceftriaxone and 5 × 10(-8) to 4 × 10(-5) mol/L of isoniazid. This proposed method was satisfactorily used in the determination of these drugs in pharmaceutical samples and human urine and serum.


Asunto(s)
Aminoacridinas/química , Ceftriaxona/análisis , Fluvoxamina/análisis , Isoniazida/análisis , Mediciones Luminiscentes/métodos , Permanganato de Potasio/química , Ceftriaxona/sangre , Ceftriaxona/orina , Fluvoxamina/sangre , Fluvoxamina/orina , Humanos , Isoniazida/sangre , Isoniazida/orina , Reproducibilidad de los Resultados , Comprimidos/análisis
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