Asunto(s)
Fallo Renal Crónico/terapia , Diálisis Peritoneal/métodos , Adulto , Anciano , Urgencias Médicas , Femenino , Humanos , Masculino , Persona de Mediana Edad , Diálisis Renal , Estados UnidosRESUMEN
This technical report presents quality control (QC) assays that can be performed in order to qualify clinical biospecimens that have been biobanked for use in research. Some QC assays are specific to a disease area. Some QC assays are specific to a particular downstream analytical platform. When such a qualification is not possible, QC assays are presented that can be performed to stratify clinical biospecimens according to their biomolecular quality.
Asunto(s)
Control de Calidad , Manejo de Especímenes/normas , Bancos de Muestras Biológicas , Investigación Biomédica/normas , Humanos , Especificidad de ÓrganosRESUMEN
The impact of shipping temperatures and preservation media used during transport of either peripheral blood mononuclear cells (PBMCs) or Jurkat cells was assessed, in view of implementing of a proficiency testing scheme on mononuclear cell viability. Samples were analyzed before and after shipment at different temperatures (ambient temperature, dry ice, and liquid nitrogen) and in different preservation media (serum with cryoprotectant, commercial cryopreservation solution, and room temperature transport medium). Sample quality was assessed by viability assays (Trypan Blue dye exclusion, flow cytometry, Cell Analysis System cell counting (CASY)), and by ELISpot functional assay. The liquid nitrogen storage and shipment were found to be the most stable conditions to preserve cell viability and functionality. However, we show that alternative high quality shipment conditions for viable cells are dry ice shipment and commercial cryopreservation solution. These were also cost-efficient shipment conditions, satisfying the requirements of a proficiency testing scheme for viable mononuclear cells. Room temperature transport medium dramatically and adversely affected the integrity of mononuclear cells.
Asunto(s)
Conservación de la Sangre/métodos , Recolección de Muestras de Sangre/métodos , Crioprotectores/farmacología , Nitrógeno/farmacología , Supervivencia Celular , Humanos , Células Jurkat , Leucocitos Mononucleares/citología , Control de Calidad , TemperaturaRESUMEN
The first version of the Standard PREanalytical Code (SPREC) was developed in 2009 by the International Society for Biological and Environmental Repositories (ISBER) Biospecimen Science Working Group to facilitate documentation and communication of the most important preanalytical quality parameters of different types of biospecimens used for research. This same Working Group has now updated the SPREC to version 2.0, presented here, so that it contains more options to allow for recent technological developments. Existing elements have been fine tuned. An interface to the Biospecimen Reporting for Improved Study Quality (BRISQ) has been defined, and informatics solutions for SPREC implementation have been developed. A glossary with SPREC-related definitions has also been added.
Asunto(s)
Bancos de Muestras Biológicas/normas , Bancos de Muestras Biológicas/organización & administración , Control de Calidad , Manejo de Especímenes/normasRESUMEN
A human papillomavirus (HPV) multiplexed competitive Luminex immunoassay first described by Opalka et al. (D. Opalka, C. E. Lachman, S. A. MacMullen, K. U. Jansen, J. F. Smith, N. Chirmule, and M. T. Esser, Clin. Diagn. Lab. Immunol. 10:108--15, 2003) was optimized and validated for use in epidemiology studies and vaccine clinical trials. Optimization increased both the analytical sensitivity and the clinical specificity of the assay to more effectively discriminate the low-titer antibody response of HPV-infected persons from noninfected individuals. The characteristics of the assay that were optimized included monoclonal antibody (MAb) specificity, scaling up the conjugation of virus-like particles (VLPs) to microspheres, VLP concentration, MAb concentration, sample matrix, sample dilution, incubation time, heat inactivation of sample sera, and detergent effects on assay buffer. The assay was automated by use of a TECAN Genesis Workstation, thus improving assay throughput, reproducibility, and operator safety. Following optimization, the assay was validated using several distinct serum panels from individuals determined to be at low and high risk for HPV infection. The validated assay was then used to determine the clinical serostatus cutoff. This high-throughput assay has proven useful for performing epidemiology studies and evaluating the efficacy of prophylactic HPV vaccines.
Asunto(s)
Anticuerpos Monoclonales/sangre , Anticuerpos Antivirales/sangre , Inmunoensayo/métodos , Papillomaviridae/inmunología , Infecciones por Papillomavirus/inmunología , Infecciones Tumorales por Virus/inmunología , Adolescente , Adulto , Niño , Epítopos/inmunología , Femenino , Humanos , Masculino , Microesferas , Papillomaviridae/clasificación , Infecciones por Papillomavirus/diagnóstico , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Factores de Tiempo , Infecciones Tumorales por Virus/diagnósticoRESUMEN
A series of 1,3,4-trisubstituted pyrrolidine CCR5 receptor antagonists containing a variety of fused heterocycles at the 4-position of the piperidine side chain has been discovered, which are orally bioavailable with potent anti-HIV activity.
Asunto(s)
Fármacos Anti-VIH/química , Antagonistas de los Receptores CCR5 , Compuestos Heterocíclicos/química , Pirrolidinas/química , Administración Oral , Animales , Fármacos Anti-VIH/farmacocinética , Células HeLa , Compuestos Heterocíclicos/farmacocinética , Humanos , Pirrolidinas/farmacocinética , Ratas , Receptores CCR5/metabolismoRESUMEN
Synthesis of analogs containing more rigid bicyclic piperidine replacements for the 4-benzyloxycarbonyl-(ethyl)amino-piperidine moiety of the CCR5 antagonist structure, 1, is described. Although similar binding affinity to the lead was achieved with some analogs they were overall less potent anti-HIV agents suggesting that other features besides CCR5 binding are required for good anti-viral activity.
Asunto(s)
Fármacos Anti-VIH/síntesis química , Antagonistas de los Receptores CCR5 , Sulfonas/síntesis química , Fármacos Anti-VIH/farmacología , Butanos/síntesis química , Butanos/farmacología , Relación Dosis-Respuesta a Droga , Concentración 50 Inhibidora , Piperidinas/síntesis química , Piperidinas/farmacología , Relación Estructura-Actividad , Sulfonas/farmacología , Virus/efectos de los fármacosRESUMEN
Efforts toward the exploration of the title compounds as CCR5 antagonists are disclosed. The basis for such work stems from the fact that cellular proliferation of HIV-1 requires the cooperative assistance of both CCR5 and CD4 receptors. The synthesis and SAR of pyrrolidineacetic acid derivatives as CCR5 antagonists displaying potent binding and antiviral properties in a HeLa cell-based HIV-1 infectivity assay are discussed.
Asunto(s)
Fármacos Anti-VIH/síntesis química , Antagonistas de los Receptores CCR5 , VIH-1/efectos de los fármacos , Pirrolidinas/síntesis química , Acetatos/química , Fármacos Anti-VIH/farmacología , Sitios de Unión , División Celular/efectos de los fármacos , Células HeLa , Humanos , Piperidinas/síntesis química , Piperidinas/farmacología , Pirrolidinas/química , Relación Estructura-ActividadRESUMEN
Cellular proliferation of HIV-1 requires the cooperative assistance of both the CCR5 and CD4 receptors. Our medicinal chemistry efforts in this area have resulted in the identification of N-alkyl piperidine sulfones as CCR5 antagonists. These compounds display potent binding and show antiviral properties in HIV-1 spread cell-based assays.
Asunto(s)
Fármacos Anti-VIH/síntesis química , Antagonistas de los Receptores CCR5 , Sulfonas/síntesis química , Fármacos Anti-VIH/farmacología , Sitios de Unión , Antígenos CD4/metabolismo , Línea Celular , Diseño de Fármacos , Humanos , Concentración 50 Inhibidora , Piperidinas/química , Receptores CCR5/metabolismo , Estereoisomerismo , Relación Estructura-Actividad , Sulfonas/farmacologíaRESUMEN
Replacement of the flexible connecting chains between the piperidine moiety and an aromatic group in previous CCR5 antagonists with heterocycles, such as pyrazole and isoxazole, provided potent CCR5 antagonists with excellent anti-HIV-1 activity in vitro. SAR studies revealed optimal placement of an unsubstituted nitrogen atom in the heterocycle to be meta to the bond connected to the 4-position of piperidine. Truncation of a benzyl group to a phenyl group afforded compounds with dramatically improved oral bioavailability, albeit with reduced activity.
Asunto(s)
Fármacos Anti-VIH/síntesis química , Fármacos Anti-VIH/farmacocinética , Antagonistas de los Receptores CCR5 , Piperidinas/síntesis química , Piperidinas/farmacocinética , Animales , Fármacos Anti-VIH/química , División Celular/efectos de los fármacos , Células HeLa , Humanos , Estructura Molecular , Piperidinas/química , Pirazoles/química , Pirazoles/farmacocinética , Ratas , Relación Estructura-ActividadRESUMEN
Modifications of the alkyl acetic acid portion and the phenyl on pyrrolidine in our lead pyrazole compound 1 afforded the isopropyl compound 9. This compound is a potent CCR5 antagonist showing good in vitro antiviral activity against HIV-1, an excellent selectivity profile, and good oral bioavailability in three animal species. During this investigation, a new method for the preparation of alpha-(pyrrolidin-1-yl)-alpha,alpha-dialkyl acetic acid from a pyrrolidine and alpha-bromo-alpha,alpha-dialkyl acetic acid using silver triflate was discovered. This allowed us to prepare compounds such as 24 and 25 for the first time. A novel Pd-mediated N-dealkylation of alpha-(pyrrolidin-1-yl)acetic acid was also uncovered.
Asunto(s)
Fármacos Anti-VIH/síntesis química , Fármacos Anti-VIH/farmacocinética , Antagonistas de los Receptores CCR5 , Piperidinas/síntesis química , Piperidinas/farmacocinética , Acetatos/química , Acetatos/farmacocinética , Administración Oral , Animales , Fármacos Anti-VIH/química , Disponibilidad Biológica , Perros , Células HeLa , Humanos , Macaca mulatta , Estructura Molecular , Monocitos/efectos de los fármacos , Piperidinas/química , Pirazoles/química , Pirazoles/farmacocinética , Ratas , Relación Estructura-ActividadRESUMEN
Extensive SAR studies in our benzylpyrazole series of CCR5 antagonists have shown that both lipophilic and hydrophilic substituents on the phenyl of the benzyl group increase antiviral potency. However, improvements in pharmacokinetic profiles were generally only observed with more lipophilic substitutions. 4-Biphenyl (51) performed the best in this regard. Highly lipophilic substituents impart undesirable ion channel activity to these CCR5 antagonists. Alkoxy substituents provide a good balance of antiviral activity, pharmacokinetic parameters, and selectivity. Compounds 42b and 42d, containing a 3,4-dimethoxy substituent, are considered the most promising improvements over parent compounds 9. They demonstrate improved antiviral activity while retaining good pharmacokinetic profile and selectivity.
Asunto(s)
Fármacos Anti-VIH/química , Fármacos Anti-VIH/farmacocinética , Antagonistas de los Receptores CCR5 , Piperidinas/química , Piperidinas/farmacocinética , Pirazoles/química , Animales , Fármacos Anti-VIH/síntesis química , Disponibilidad Biológica , Perros , Células HeLa , Humanos , Estructura Molecular , Monocitos/efectos de los fármacos , Piperidinas/síntesis química , Pirazoles/farmacocinética , Ratas , Relación Estructura-ActividadRESUMEN
[reaction: see text] A novel approach to alpha,alpha-disubstituted-beta-amino acids (beta(2,2)-amino acids) was employed in the synthesis of a series of 3-(pyrrolidin-1-yl)propionic acids possessing high affinity for the CCR5 receptor and potent anti-HIV activity. The rat pharmacokinetics for these new analogues featured higher bioavailabilities and lower rates of clearance as compared to cyclopentane 1.
Asunto(s)
Fármacos Anti-VIH/farmacología , Antagonistas de los Receptores CCR5 , Propionatos/farmacología , Pirrolidinas/farmacología , Fármacos Anti-VIH/farmacocinética , Disponibilidad Biológica , Propionatos/farmacocinética , Pirrolidinas/farmacocinéticaRESUMEN
A new class of 4-(aminoheterocycle)piperidine derived 1,3,4 trisubstituted pyrrolidine CCR5 antagonists is reported. Compound 4a is shown to have good binding affinity (1.8 nM) and antiviral activity in PBMC's (IC(95)=50 nM). Compound 4a also has improved PK properties relative to 1.
Asunto(s)
Antagonistas de los Receptores CCR5 , Piperidinas/síntesis química , Piperidinas/farmacología , Pirrolidinas/síntesis química , Pirrolidinas/farmacología , Animales , Fármacos Anti-VIH/síntesis química , Células CHO , Quimiocina CCL4 , Cricetinae , Semivida , Células HeLa , Humanos , Enlace de Hidrógeno , Proteínas Inflamatorias de Macrófagos/metabolismo , Piperidinas/farmacocinética , Pirrolidinas/farmacocinética , RatasRESUMEN
The 4-(3-phenylprop-1-yl)piperidine moiety of the 1,3,4-trisubstituted pyrrolidine CCR5 antagonist 1 was modified with electron deficient aromatics as well as replacement of the benzylic methylene with sulfones, gem-difluoromethylenes and alcohols in an effort to balance the antiviral potency with reasonable pharmacokinetics.
Asunto(s)
Fármacos Anti-VIH/síntesis química , Antagonistas de los Receptores CCR5 , Pirrolidinas/farmacocinética , Animales , Fármacos Anti-VIH/farmacocinética , Fármacos Anti-VIH/farmacología , Perros , Semivida , Humanos , Leucocitos Mononucleares , Macaca mulatta , Tasa de Depuración Metabólica , Piperidinas/química , Pirrolidinas/síntesis química , Pirrolidinas/farmacología , Ensayo de Unión Radioligante , Ratas , Relación Estructura-Actividad , Células Tumorales CultivadasRESUMEN
Incorporation of acidic functional groups into a lead CCR5 antagonist identified from a targeted combinatorial library resulted in compounds with enhanced anti-HIV-1 activity and attenuated L-type calcium channel affinity.
Asunto(s)
Fármacos Anti-VIH/síntesis química , Fármacos Anti-VIH/farmacología , Antagonistas de los Receptores CCR5 , VIH-1/efectos de los fármacos , Animales , Fármacos Anti-VIH/farmacocinética , Células CHO , Canales de Calcio Tipo L/efectos de los fármacos , Fenómenos Químicos , Química Física , Quimiocina CCL4 , Cricetinae , Semivida , Células HeLa , Humanos , Proteínas Inflamatorias de Macrófagos/antagonistas & inhibidores , Ratas , Receptores CCR5/química , Relación Estructura-ActividadRESUMEN
A series of alpha-(pyrrolidin-1-yl)acetic acids is presented as selective and potent antivirals against HIV. Several of the pyrrolidine zwitterions demonstrated reasonable in vitro properties, enhanced antiviral activities and improved pharmacokinetic profiles over pyrrolidine 1.
Asunto(s)
Fármacos Anti-VIH/síntesis química , Fármacos Anti-VIH/farmacología , Antagonistas de los Receptores CCR5 , VIH-1/efectos de los fármacos , Pirrolidinas/síntesis química , Pirrolidinas/farmacología , Animales , Fármacos Anti-VIH/farmacocinética , Células CHO , Permeabilidad de la Membrana Celular , Fenómenos Químicos , Química Física , Quimiocina CCL4 , Cricetinae , Células HeLa , Humanos , Indicadores y Reactivos , Proteínas Inflamatorias de Macrófagos/antagonistas & inhibidores , Pirrolidinas/farmacocinética , Ratas , Relación Estructura-ActividadRESUMEN
A series of CCR5 antagonists containing bicyclic isoxazolidines was generated through a nitrone mediated cycloaddition with olefins bearing the preferred pharmacophores previously described. Potent antagonists (3 and 16) were generated with enhanced affinity for the CCR5 receptor while maintaining antiviral activity against HIV.