RESUMEN
BACKGROUND: The study of speciation has expanded with the increasing availability and affordability of high-resolution genomic data. How the genome evolves throughout the process of divergence and which regions of the genome are responsible for causing and maintaining that divergence have been central questions in recent work. Here, we use three pairs of species from the recently diverged bee hummingbird clade to investigate differences in the genome at different stages of speciation, using divergence times as a proxy for the speciation continuum. RESULTS: Population measures of relative differentiation between hybridizing species reveal that different chromosome types diverge at different stages of speciation. Using FST as our relative measure of differentiation we found that the sex chromosome shows signs of divergence early in speciation. Next, small autosomes (microchromosomes) accumulate highly diverged genomic regions, while the large autosomes (macrochromosomes) accumulate genomic regions of divergence at a later stage of speciation. CONCLUSIONS: Our finding that genomic windows of elevated FST accumulate on small autosomes earlier in speciation than on larger autosomes is counter to the prediction that FST increases with size of chromosome (i.e. with decreased recombination rate), and is not represented when weighted average FST per chromosome is compared with chromosome size. The results of this study suggest that multiple chromosome characteristics such as recombination rate and gene density combine to influence the genomic locations of signatures of divergence.
Asunto(s)
Evolución Biológica , Aves/clasificación , Especiación Genética , Animales , Cromosomas/genética , Genoma , Genómica , Hibridación Genética , Cromosomas Sexuales/genéticaRESUMEN
Respiratory syncytial virus (RSV) is the most common cause of lower respiratory tract infections worldwide, yet no effective vaccine or antiviral treatment is available. Here we report the discovery and initial development of RSV604, a novel benzodiazepine with submicromolar anti-RSV activity. It proved to be equipotent against all clinical isolates tested of both the A and B subtypes of the virus. The compound has a low rate of in vitro resistance development. Sequencing revealed that the resistant virus had mutations within the nucleocapsid protein. This is a novel mechanism of action for anti-RSV compounds. In a three-dimensional human airway epithelial cell model, RSV604 was able to pass from the basolateral side of the epithelium effectively to inhibit virus replication after mucosal inoculation. RSV604, which is currently in phase II clinical trials, represents the first in a new class of RSV inhibitors and may have significant potential for the effective treatment of RSV disease.
Asunto(s)
Antivirales/farmacología , Benzodiazepinonas/farmacología , Compuestos de Fenilurea/farmacología , Virus Sincitiales Respiratorios/efectos de los fármacos , Secuencia de Aminoácidos , Antivirales/síntesis química , Benzodiazepinonas/síntesis química , Línea Celular , Fenómenos Químicos , Química Física , Efecto Citopatogénico Viral , Relación Dosis-Respuesta a Droga , Farmacorresistencia Viral/genética , Células Epiteliales/efectos de los fármacos , Células Epiteliales/virología , Humanos , Datos de Secuencia Molecular , Mutagénesis Sitio-Dirigida , Mutación , Proteínas de la Nucleocápside/efectos de los fármacos , Compuestos de Fenilurea/síntesis química , Virus Sincitiales Respiratorios/genética , Sales de Tetrazolio , Replicación Viral/efectos de los fármacosRESUMEN
Respiratory syncytial virus (RSV) is the cause of one-fifth of all lower respiratory tract infections worldwide and is increasingly being recognized as representing a serious threat to patient groups with poorly functioning or immature immune systems. Racemic 1,4-benzodiazepines show potent anti-RSV activity in vitro. Anti-RSV evaluation of 3-position R- and S-benzodiazepine enantiomers and subsequent optimization of this series resulted in selection of a clinical candidate. Antiviral activity was found to reside mainly in the S-enantiomer, and the R-enantiomers were consistently less active against RSV. Analogues of 1,4-(S)-benzodiazepine were synthesized as part of the lead optimization program at Arrow and tested in the XTT assay. From this exercise, (S)-1-(2-fluorophenyl)-3-(2-oxo-5-phenyl-2,3-dihydro-1H-benzo[e][1,4]-diazepin-3-yl)-urea, 17b (RSV-604) was identified as a clinical candidate, exhibiting potent anti-RSV activity in the XTT assay, which was confirmed in secondary assays. Compound 17b also possessed a good pharmacokinetic profile and has now progressed into the clinic.
Asunto(s)
Antivirales/síntesis química , Benzodiazepinas/síntesis química , Benzodiazepinonas/síntesis química , Compuestos de Fenilurea/síntesis química , Virus Sincitiales Respiratorios/efectos de los fármacos , Animales , Antivirales/farmacocinética , Antivirales/farmacología , Benzodiazepinas/farmacocinética , Benzodiazepinas/farmacología , Benzodiazepinonas/farmacocinética , Benzodiazepinonas/farmacología , Línea Celular Tumoral , Cristalografía por Rayos X , Perros , Ensayo de Inmunoadsorción Enzimática , Humanos , Técnicas In Vitro , Microsomas/metabolismo , Estructura Molecular , Compuestos de Fenilurea/farmacocinética , Compuestos de Fenilurea/farmacología , Ratas , Estereoisomerismo , Relación Estructura-Actividad , Ensayo de Placa ViralRESUMEN
The alpha-FR has been reported to be overexpressed in many carcinomas, in particular those of the ovary and uterus. The high expression of alpha-FR in some tumours compared with normal tissues has been exploited over the last decade for folate-mediated targeting of macromolecules, anticancer drugs, imaging agents and nucleic acids to cancer cells. CB300638, a cyclopenta[g]quinazoline-based inhibitor of thymidylate synthase (TS), has been reported to have high affinity for the receptor and selectivity for alpha-FR overexpressing tumour cell lines. In this study, the structural features of the molecule, in particular modifications at the 2-position, have been investigated with respect to TS inhibition, affinity for the alpha-FR and reduced folate carrier (RFC) and activity in A431-FBP cells (transfected with human alpha-FR) compared with neo-transfected A431 cells. Compounds 1a,b, 2a,b and 3a,b were synthesised utilising multistep sequences. It was found that the 2-substituent does not affect the affinity for the alpha-FR; however, it greatly affects selectivity for A431-FBP cells, and suggests that there are factors other than TS inhibition and alpha-FR affinity that are important for the activity of these compounds. Compound 2b (2-CH2OH derivative) displayed the highest selectivity for the A431-FBP cells compared with A431 cells.
Asunto(s)
Proteínas Portadoras/antagonistas & inhibidores , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Quinazolinas/química , Quinazolinas/farmacología , Receptores de Superficie Celular/antagonistas & inhibidores , Timidilato Sintasa/antagonistas & inhibidores , Proteínas Portadoras/genética , Proteínas Portadoras/metabolismo , Línea Celular , Ciclopentanos/química , Ciclopentanos/farmacología , Inhibidores Enzimáticos/síntesis química , Receptores de Folato Anclados a GPI , Humanos , Técnicas In Vitro , Quinazolinas/síntesis química , Receptores de Superficie Celular/genética , Receptores de Superficie Celular/metabolismo , Proteínas Recombinantes/antagonistas & inhibidores , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Relación Estructura-Actividad , TransfecciónRESUMEN
Respiratory syncytial virus (RSV) is the cause of one-fifth of all lower respiratory tract infections worldwide and is increasingly being recognized as a serious threat to patient groups with poorly functioning immune systems. Our approach to finding a novel inhibitor of this virus was to screen a 20 000-member diverse library in a whole cell XTT assay. Parallel assays were carried out in the absence of virus in order to quantify any associated cell toxicity. This identified 100 compounds with IC(50)'s less than 50 muM. A-33903 (18), a 1,4-benzodiazepine analogue, was chosen as the starting point for lead optimization. This molecule was moderately active and demonstrated good pharmacokinetic properties. The most potent compounds identified from this work were A-58568 (47), A-58569 (44), and A-62066 (46), where modifications to the aromatic substitution enhanced potency, and A-58175 (42), where the amide linker was modified.
Asunto(s)
Antivirales/síntesis química , Benzodiazepinas/síntesis química , Virus Sincitiales Respiratorios/efectos de los fármacos , Animales , Antivirales/farmacocinética , Antivirales/farmacología , Benzodiazepinas/química , Benzodiazepinas/farmacología , Línea Celular Tumoral , Técnicas Químicas Combinatorias , Ensayo de Inmunoadsorción Enzimática , Ratas , Ratas Sprague-Dawley , Relación Estructura-Actividad , Ensayo de Placa ViralRESUMEN
BGC 945 is a cyclopenta[g]quinazoline-based, thymidylate synthase inhibitor specifically transported into alpha-folate receptor (alpha-FR)-overexpressing tumors. Affinity of BGC 945 for the alpha-FR is 70% of the high-affinity ligand folic acid. In contrast to conventional antifolates, BGC 945 has low affinity for the widely expressed reduced-folate carrier (RFC). The K(i) for isolated thymidylate synthase is 1.2 nmol/L and the IC(50) for inhibition of the growth of alpha-FR-negative mouse L1210 or human A431 cells is approximately 7 micromol/L. In contrast, BGC 945 is highly potent in a range of alpha-FR-overexpressing human tumor cell lines (IC(50) approximately 1-300 nmol/L). Pharmacokinetic variables measured following i.v. injection of 100 mg/kg BGC 945 to KB tumor-bearing mice showed rapid plasma clearance (0.021 L/h) and tissue distribution. The terminal half-lives in plasma, liver, kidney, spleen, and tumor were 2, 0.6, 5, 21, and 28 hours, respectively. Tumor BGC 945 concentration at 24 hours was approximately 1 nmol/g tissue, at least 10-fold higher than that in plasma or normal tissues. Inhibition of thymidylate synthase in tissues leads to increased incorporation of 5-[(125)I]-iodo-2'-deoxyuridine ([(125)I]dUrd) into DNA. Forty-eight hours after injection of 100 mg/kg 6RS-BGC 945 ([(125)I]dUrd injected at 24 hours), tumor was the only tissue with incorporation above control level (6-fold). The RFC-mediated thymidylate synthase inhibitor plevitrexed also increased uptake of [(125)I]dUrd in tumor (10-fold) but, in contrast, also caused increased incorporation in other normal tissues such as spleen and small bowel (4.5- and 4.6-fold, respectively). These data suggest that BGC 945 selectively inhibits thymidylate synthase in alpha-FR-overexpressing tumors and should cause minimal toxicity to humans at therapeutic doses.
Asunto(s)
Proteínas Portadoras/metabolismo , Inhibidores Enzimáticos/farmacología , Quinazolinas/farmacología , Receptores de Superficie Celular/metabolismo , Timidilato Sintasa/antagonistas & inhibidores , Animales , Apoptosis/efectos de los fármacos , Transporte Biológico , Proliferación Celular/efectos de los fármacos , Coriocarcinoma/tratamiento farmacológico , Coriocarcinoma/enzimología , Inhibidores Enzimáticos/farmacocinética , Femenino , Receptores de Folato Anclados a GPI , Ácido Fólico/metabolismo , Humanos , Idoxuridina/metabolismo , Radioisótopos de Yodo , Leucemia L1210/tratamiento farmacológico , Leucemia L1210/enzimología , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/enzimología , Proteínas de Transporte de Membrana , Ratones , Ratones Desnudos , Neoplasias Ováricas/tratamiento farmacológico , Neoplasias Ováricas/enzimología , Quinazolinas/farmacocinética , Proteína Portadora de Folato Reducido , Distribución Tisular , Trasplante Heterólogo , Células Tumorales CultivadasRESUMEN
A new route to compound 3 (4-[N-(6RS)-2-methyl-4-oxo-3,4,7,8-tetrahydro-6H-cyclopenta[g]quinazolin-6-yl]-N-(prop-2-ynyl)amino]benzoic acid), a crucial intermediate for the synthesis of potent inhibitors of thymidylate synthase (TS), is described. In this sequence the C6-N10 bond was constructed first, by the reductive amination of 5-acetamido-6-bromoindan-1-one 6 with tert-butyl 4-aminobenzoate, then the cyclopenta[g]quinazolinone ring was formed and the propargyl group was introduced on the N10-position using the (propargyl)Co2(CO)6+ complex as the electrophilic propargyl reagent.