RESUMEN
A thorough investigation into the mechanism of the reaction of 3-methylpentanoic acid and Meldrum's acid using online NMR spectroscopy is reported. This study is an expansion of a previous analysis of this chemical transformation in the synthesis of an active pharmaceutical ingredient imagabalin. The 3-methylpentanoic acid analogue reveals similar behavior under the reaction conditions. Online NMR spectroscopy and offline characterization experiments reveal new information about the mechanism, providing conclusive spectroscopic evidence for the previously hypothesized dimer anhydride intermediate species 3-methylpentanoic anhydride as a productive intermediate. The presence of an acyl chloride intermediate species, 3-methylpentanoyl chloride, is also revealed for the first time in this synthesis. Copyright © 2015 John Wiley & Sons, Ltd.
RESUMEN
In this paper we describe the development of a 5 mm NMR flow tube that can be used in a standard 5 mm NMR probe, enabling the user to conduct experiments on flowing samples or, more specifically, on flowing reaction mixtures. This enables reaction monitoring or kinetic experiments to be conducted by flowing reaction mixtures from a reaction vessel to detection in the coil area of the NMR, without the need for a specialized flow NMR probe. One of the key benefits of this flow tube is that it provides flexibility to be used across a range of available spectrometers of varying magnetic field strengths with a standard 5 mm probe setup. The applicability of this flow tube to reaction monitoring is demonstrated using the reaction of p-phenylenediamine and isobutyraldehyde to form the diimine product.
Asunto(s)
Técnicas de Química Analítica/instrumentación , Técnicas de Química Analítica/métodos , Espectroscopía de Resonancia Magnética/instrumentación , Aldehídos/química , Bencidinas/química , Fenilendiaminas/químicaRESUMEN
Detector response is not always equivalent between detectors or instrument types. Factors that impact detector response include molecular structure and detection wavelength. In liquid chromatography (LC), ultraviolet (UV) is often the primary detector; however, without determination of UV response factors for each analyte, chromatographic results are reported on an area percent rather than a weight percent. In extreme cases, response factors can differ by several orders of magnitude for structurally dissimilar compounds, making the uncalibrated data useless for quantitative applications. While impurity reference standards are normally used to calculate UV relative response factors (RRFs), reference standards of reaction mixture components are typically not available during route scouting or in the early stages of process development. Here, we describe an approach to establish RRFs from a single experiment using both online nuclear magnetic resonance (NMR) and LC. NMR is used as a mass detector from which a UV response factor can be determined to correct the high performance liquid chromatography (HPLC) data. Online reaction monitoring using simultaneous NMR and HPLC provides a platform to expedite the development and understanding of pharmaceutical reaction processes. Ultimately, the knowledge provided by a structurally information rich technique such as NMR can be correlated with more prevalent and mobile instrumentation [e.g., LC, mid-infrared spectrometers (MIR)] for additional routine process understanding and optimization.
Asunto(s)
Cromatografía Líquida de Alta Presión/instrumentación , Espectroscopía de Resonancia Magnética/instrumentación , Sistemas en Línea/instrumentación , Preparaciones Farmacéuticas/química , Tecnología Farmacéutica/instrumentación , Estructura MolecularRESUMEN
A contracted ring degradation product, WYE-120318 (compound 2), was discovered during the development phase for methylnaltrexone bromide (compound 1) drug substance. The compound was isolated by high-performance liquid chromatography fractionation, and its structure was determined by spectroscopic data analyses. WYE-120318 is formed from methylnaltrexone through a benzyl-benzilic acid type rearrangement reaction to yield an α-hydroxy-cyclopentanecarboxylic acid substructure. The proposed structure and the formation mechanism are confirmed by the synthesis of WYE-120318 from methylnaltrexone (compound 1). A similar benzyl-benzilic acid type rearrangement reaction can be envisioned as the biological origin of remisporine A (compound 3), a naturally occurring cyclopentadienyl compound that autocatalytically dimerizes to remisporine B (compound 4). The structure of remisporine A was deduced from its dimer 4. Coniothyione (compound 5) can be considered as the first example of a stable natural product bearing the remisporine A skeleton. However, the regiochemistry of the chlorosubstitution in the coniothyrione structure needs to be revised to compound 6 on the basis of the nuclear magnetic resonance data and biogenesis analysis.
Asunto(s)
Cromonas/química , Naltrexona/análogos & derivados , Catálisis , Cromatografía Líquida de Alta Presión , Dimerización , Espectroscopía de Resonancia Magnética , Naltrexona/síntesis química , Naltrexona/química , Compuestos de Amonio Cuaternario/química , EstereoisomerismoRESUMEN
A screening program for bioactive compounds from marine cyanobacteria led to the isolation of jamaicamides A-C. Jamaicamide A is a novel and highly functionalized lipopeptide containing an alkynyl bromide, vinyl chloride, beta-methoxy eneone system, and pyrrolinone ring. The jamaicamides show sodium channelblocking activity and fish toxicity. Precursor feeding to jamaicamide-producing cultures mapped out the series of acetate and amino acid residues and helped develop an effective cloning strategy for the biosynthetic gene cluster. The 58 kbp gene cluster is composed of 17 open reading frames that show an exact colinearity with their expected utilization. A novel cassette of genes appears to form a pendent carbon atom possessing the vinyl chloride functionality; at its core this contains an HMG-CoA synthase-like motif, giving insight into the mechanism by which this functional group is created.
Asunto(s)
Amidas/química , Cianobacterias/química , Toxinas Marinas/química , Neurotoxinas/química , Péptidos/química , Pirrolidinonas/química , Amidas/aislamiento & purificación , Amidas/farmacología , Animales , Secuencia de Bases , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Cianobacterias/genética , Cianobacterias/metabolismo , Humanos , Lipopéptidos , Lipoproteínas/química , Lipoproteínas/aislamiento & purificación , Lipoproteínas/farmacología , Toxinas Marinas/aislamiento & purificación , Toxinas Marinas/farmacología , Ratones , Datos de Secuencia Molecular , Estructura Molecular , Familia de Multigenes , Neurotoxinas/aislamiento & purificación , Neurotoxinas/farmacología , Péptidos/aislamiento & purificación , Péptidos/farmacología , Estructura Terciaria de Proteína , Pirrolidinonas/aislamiento & purificación , Pirrolidinonas/farmacología , Canales de Sodio/efectos de los fármacos , Canales de Sodio/fisiologíaRESUMEN
Phytochemical examination of a Papua New Guinea collection of Lyngbya majuscula resulted in the discovery of wewakazole (1), a novel cyclic dodecapeptide containing an unprecedented six five-membered heterocycles. Multiple NMR experiments and MS/MS data were required to assemble its planar structure because of its extensively signal-overlapped NMR spectra. In particular, a 1D HMBC was utilized to orient a three amino acid fragment that could not be placed by standard spectroscopic methods. [structure--see text]
Asunto(s)
Péptidos Cíclicos/química , Péptidos Cíclicos/aislamiento & purificación , Algas Marinas/química , Espectroscopía de Resonancia Magnética , Espectrometría de Masas , Estructura Molecular , Papúa Nueva GuineaRESUMEN
Here we report the detailed measurement of long-range heteronuclear spin-spin coupling constants, especially 2, 3JCH spin-spin couplings for eight different cyclopentane derivatives. These 2, 3JCH constants were shown to be a useful tool in the determination of the relative stereochemistry in these rings. The coupling constant measurements reported here are based on two different experiments: a 2D heteronuclear correlation experiment named G-BIRDR, X-CPMG-HSQMBC and the 2D-coupled gHSQC {1H-13C} experiment
Asunto(s)
Ciclopentanos/química , Espectroscopía de Resonancia Magnética , Espectroscopía de Resonancia Magnética/normas , Estructura Molecular , Estándares de ReferenciaRESUMEN
The application of Chiral Technology, or the (extensive) use of techniques or tools for the determination of absolute stereochemistry and the enantiomeric or chiral separation of racemic small molecule potential lead compounds, has been critical to successfully discovering and developing chiral drugs in the pharmaceutical industry. This has been due to the rapid increase over the past 10-15 years in potential drug candidates containing one or more asymmetric centers. Based on the experiences of one pharmaceutical company, a summary of the establishment of a Chiral Technology toolbox, including the implementation of known tools as well as the design, development, and implementation of new Chiral Technology tools, is provided.
Asunto(s)
Química Farmacéutica/métodos , Industria Farmacéutica/métodos , Estereoisomerismo , Cromatografía/métodos , Dicroismo Circular , Diseño de Fármacos , Electrones , Humanos , Espectroscopía de Resonancia Magnética/métodos , Modelos Químicos , Conformación Molecular , Sales (Química)/farmacología , Tecnología Farmacéutica/métodosRESUMEN
In vitro drug metabolism studies during the early drug discovery stage are becoming increasingly important. With the increasing demand for high throughput and quick turnaround time for in vitro metabolism studies, however, careful examination of the results and proper design of the experiments are still crucial. In this communication, we report the identification and mechanism of formation of a novel metabonate from incubations of a diamine-containing compound with liver microsomes. The metabonate appeared to be the major product, and its formation was NADPH- and microsomal protein-dependent. Liquid chromatography/mass spectrometry and NMR analysis of the metabonate indicated an extra carbon and unusual formation of an imidazolidine ring. Further studies revealed that this metabonate was not a true biotransformation product from the diamine compound itself in the microsomal incubation, but rather a product resulting from a condensation reaction between the compound and a metabolite of the solvent (alcohol) used in the incubation. When the microsomal incubations contained a small amount of methanol or ethanol as solvent, the alcohols were metabolized to formaldehyde or acetaldehyde, which then condensed with the diamine compound through an imine intermediate to form the metabonate. The compound itself was metabolically stable in vitro when acetonitrile or dimethyl sulfoxide was used as solvent. During the study of in vitro microsomal stability and metabolite identification of amine-containing compounds, the use of alcohol as solvent should be avoided.
Asunto(s)
Artefactos , Evaluación Preclínica de Medicamentos , Etilenodiaminas/metabolismo , Microsomas Hepáticos/metabolismo , Animales , Etanol/química , Etilenodiaminas/química , Humanos , Técnicas In Vitro , Espectroscopía de Resonancia Magnética , Espectrometría de Masas , Metanol/química , Estructura Molecular , Ratas , Solventes/químicaRESUMEN
Detailed measurements of long-range heteronuclear spin-spin coupling constants, especially (2, 3)J(CH) spin-spin couplings for various cyclopentane derivatives, are reported. The measurements are based on a 2D heteronuclear correlation experiment named G-BIRD(R, X)-CPMG-HSQMBC.
Asunto(s)
Ciclopentanos/química , Espectroscopía de Resonancia Magnética , Ciclopentanos/clasificación , Ciclopentanos/normas , Estructura Molecular , Estándares de ReferenciaRESUMEN
Aqueous phosphoric acid (85 wt %) is an effective, environmentally benign reagent for the deprotection of tert-butyl carbamates, tert-butyl esters, and tert-butyl ethers. The reaction conditions are mild and offer good selectivity in the presence of other acid-sensitive groups, including CBZ carbamates, azetidine, benzyl and methyl esters, TBDMS, and methyl phenyl ethers. The mildness of the reaction is further demonstrated in the synthesis of clarithromycin derivative, in which a tert-butyl ester is removed in the presence of cyclic carbamate, lactone, ketal, acetate ester, and epimerizable methyl ketone functionalities. The reaction preserves the stereochemical integrity of the substrates. The reactions are high yielding, and the workup is convenient.
Asunto(s)
Carbamatos/química , Ésteres/química , Éteres/química , Indicadores y Reactivos/química , Ácidos Fosfóricos/química , Espectroscopía de Resonancia Magnética , AguaRESUMEN
A glycosidic macrolide, lyngbouilloside (1), was characterized from the marine cyanobacterium Lyngbya bouillonii collected from Papua New Guinea. Its structure was elucidated by spectroscopic analysis and chemical derivatization. Relative stereochemistry was deduced from homonuclear and heteronuclear coupling constants as well as NOE information. Lyngbouilloside was modestly cytotoxic to neuroblastoma cells (IC(50) value of 17 microM).
Asunto(s)
Anabaena/química , Antineoplásicos/aislamiento & purificación , Glicósidos/aislamiento & purificación , Macrólidos/aislamiento & purificación , Animales , Antineoplásicos/química , Antineoplásicos/farmacología , Cromatografía Liquida , Ensayos de Selección de Medicamentos Antitumorales , Glicósidos/química , Glicósidos/farmacología , Concentración 50 Inhibidora , Macrólidos/química , Macrólidos/farmacología , Ratones , Estructura Molecular , Neuroblastoma , Nueva Guinea , Resonancia Magnética Nuclear Biomolecular , Estereoisomerismo , Células Tumorales Cultivadas/efectos de los fármacosRESUMEN
Bruchins are 3-hydroxypropanoate esters of long-chain alpha,omega-diols from pea weevils and cowpea weevils that have been shown to initiate callus formation on pea pods at extremely low application rates. Synthetic analogs have been prepared and examined to evaluate structural requirements for inducing this unusual neoplastic response. Chain length (optimum length C22-C24) is important, whereas unsaturation within the chain is relatively unimportant. Difunctionality is required for maximum activity, but the alpha,omega-diols themselves are inactive. Most critical is the ester portion(s) of the molecules; 3-hydroxypropanoate esters are far more active than any analogs examined.
Asunto(s)
Escarabajos/química , Ácido Láctico/análogos & derivados , Ácido Láctico/farmacología , Animales , Ésteres , Ácido Láctico/agonistas , Ácido Láctico/química , Mitógenos , Estructura Molecular , Pisum sativum , Plantas Comestibles , Relación Estructura-ActividadRESUMEN
Hectochlorin (1) was isolated from marine isolates of Lyngbya majuscula collected from Hector Bay, Jamaica, and Boca del Drago Beach, Bocas del Toro, Panama. The planar structure was deduced by one- and two-dimensional NMR spectroscopy. X-ray crystallography was used to determine the absolute stereochemistry of hectochlorin as 2S,3S,14S,22S. Hectochlorin is equipotent to jasplakinolide (5) in its ability to promote actin polymerization, but unlike jasplakinolide, is unable to displace a fluorescent phalloidin analogue from polymerized actin. In addition, hectochlorin shows both a unique profile of cytotoxicity by the COMPARE algorithm and potent inhibitory activity toward the fungus Candida albicans. Structurally, hectochlorin resembles dolabellin and the recently reported lyngbyabellin class of compounds.
Asunto(s)
Actinas/efectos de los fármacos , Actinas/metabolismo , Antibacterianos/aislamiento & purificación , Antifúngicos/aislamiento & purificación , Cianobacterias/química , Depsipéptidos , Lactonas/aislamiento & purificación , Tiazoles/aislamiento & purificación , Antibacterianos/química , Antibacterianos/farmacología , Antifúngicos/química , Antifúngicos/farmacología , Candida albicans/efectos de los fármacos , Cristalografía por Rayos X , Fluoresceína-5-Isotiocianato , Concentración 50 Inhibidora , Jamaica , Lactonas/química , Lactonas/farmacología , Toxinas de Lyngbya/química , Toxinas de Lyngbya/aislamiento & purificación , Toxinas de Lyngbya/farmacología , Conformación Molecular , Estructura Molecular , Resonancia Magnética Nuclear Biomolecular , Panamá , Péptidos Cíclicos/química , Péptidos Cíclicos/aislamiento & purificación , Péptidos Cíclicos/farmacología , Tiazoles/química , Tiazoles/farmacologíaRESUMEN
Marine natural products provide a rich source of chemical diversity that can be used to design and develop new, potentially useful therapeutic agents. We report here that scytonemin, a pigment isolated from cyanobacteria, is the first described small molecule inhibitor of human polo-like kinase, a serine/threonine kinase that plays an integral role in regulating the G(2)/M transition in the cell cycle. Scytonemin inhibited polo-like kinase 1 activity in a concentration-dependent manner with an IC(50) of 2 microM against the recombinant enzyme. Biochemical analysis showed that scytonemin reduced GST-polo-like kinase 1 activity in a time-independent fashion, suggesting reversibility, and with a mixed-competition mechanism with respect to ATP. Although scytonemin was less potent against protein kinase A and Tie2, a tyrosine kinase, it did inhibit other cell cycle-regulatory kinases like Myt1, checkpoint kinase 1, cyclin-dependent kinase 1/cyclin B, and protein kinase Cbeta2 with IC(50) values similar to that seen for polo-like kinase 1. Consistent with these effects, scytonemin effectively attenuated, without chemical toxicity, the growth factor- or mitogen-induced proliferation of three cell types commonly implicated in inflammatory hyperproliferation. Similarly, scytonemin (up to 10 microM) was not cytotoxic to nonproliferating endotoxin-stimulated human monocytes. In addition, Jurkat T cells treated with scytonemin were induced to undergo apoptosis in a non-cell cycle-dependent manner consistent with its activities on multiple kinases. Here we propose that scytonemin's dimeric structure, unique among natural products, may be a valuable template for the development of more potent and selective kinase inhibitors used for the treatment of hyperproliferative disorders.