RESUMEN
The previously accepted structure of the marine toxin azaspiracid-3 is revised based upon an original convergent and stereoselective total synthesis of the natural product. The development of a structural revision hypothesis, its testing, and corroboration are reported. Synthetic (6R,10R,13R,14R,16R,17R,19S,20S,21R,24S,25S,28S,30S,32R, 33R,34R,36S,37S,39R)-azaspiracid-3 chromatographically and spectroscopically matched naturally occurring azaspiracid-3, whereas the previously assigned 20R epimer did not.
Asunto(s)
Productos Biológicos/química , Productos Biológicos/síntesis química , Furanos/química , Furanos/síntesis química , Piranos/química , Piranos/síntesis química , Espectroscopía de Resonancia Magnética con Carbono-13 , Cromatografía Liquida , Espectrometría de Masas , Estructura Molecular , Oxidación-Reducción , Espectroscopía de Protones por Resonancia Magnética , EstereoisomerismoRESUMEN
A convergent and stereoselective total synthesis of the previously assigned structure of azaspiracid-3 has been achieved by a late-stage Nozaki-Hiyama-Kishi coupling to form the C21-C22 bond with the C20 configuration unambiguously established from l-(+)-tartaric acid. Postcoupling steps involved oxidation to an ynone, modified Stryker reduction of the alkyne, global deprotection, and oxidation of the resulting C1 primary alcohol to the carboxylic acid. The synthetic product matched naturally occurring azaspiracid-3 by mass spectrometry, but differed both chromatographically and spectroscopically.
Asunto(s)
Productos Biológicos/química , Furanos/síntesis química , Piranos/síntesis química , Espectroscopía de Resonancia Magnética con Carbono-13 , Cromatografía Liquida , Furanos/química , Estructura Molecular , Oxidación-Reducción , Espectroscopía de Protones por Resonancia Magnética , Piranos/química , Estereoisomerismo , Espectrometría de Masas en TándemRESUMEN
Identifying an enzyme's substrates is essential to understand its function, yet it remains challenging. A fundamental impediment is the transient interactions between an enzyme and its substrates. In contrast, tight binding is often observed for multisubstrate-adduct inhibitors due to synergistic interactions. Extending this venerable concept to enzyme-catalyzed in situ adduct formation, unknown substrates were affinity-captured by an S-adenosyl-methionine (AdoMet, SAM)-dependent methyltransferase (MTase). Specifically, the electrophilic methyl sulfonium (alkyl donor) in AdoMet is replaced with a vinyl sulfonium (Michael acceptor) in S-adenosyl-vinthionine (AdoVin). Via an addition reaction, AdoVin and the nucleophilic substrate form a covalent bisubstrate-adduct tightly complexed with thiopurine MTase (2.1.1.67). As such, an unknown substrate was readily identified from crude cell lysates. Moreover, this approach is applicable to other systems, even if the enzyme is unknown.
Asunto(s)
Etionina/análogos & derivados , Metiltransferasas/química , Metiltransferasas/metabolismo , S-Adenosilmetionina/química , S-Adenosilmetionina/metabolismo , Cromatografía Líquida de Alta Presión , Química Clic , Etionina/química , Etionina/metabolismo , Humanos , Espectrofotometría Ultravioleta , Especificidad por SustratoRESUMEN
Here, we describe the synthesis of the single amino acid chelator DOTAlaP and four of its derivatives. The corresponding gadolinium(III) complexes were investigated for their kinetic inertness, relaxometric properties at a range of fields and temperatures, water exchange rate, and interaction with human serum albumin (HSA). Derivatives with one inner-sphere water (q = 1) were determined to have a mean water residency time between 8 and 6 ns in phoshate-buffered saline at 37 °C. The corresponding europium complexes were also formed and used to obtain information on the hydration number of the corresponding coordination complexes. Two complexes capable of binding HSA were also synthesized, of which one, Gd(5b), contains no inner-sphere water, while the other derivative, Gd(4b), is a mixture of ca. 15% q =1 and 85% q = 0. In the presence of HSA, the latter displayed a very short mean water residency time (τM(310) = 2.4 ns) and enhanced relaxivity at intermediate and high fields. The kinetic inertness of Gd(4b) with respect to complex dissociation was decreased compared to its DOTAla analogue but still 100-fold more inert than [Gd(BOPTA)(H2O)](2-). Magnetic resonance imaging in mice showed that Gd(4b) was able to provide 38% better vessel to muscle contrast compared to the clinically used HSA binding agent MS-325.
Asunto(s)
Medios de Contraste/química , Gadolinio/química , Compuestos Heterocíclicos con 1 Anillo/química , Agua/química , Albúminas/química , Animales , Quelantes/química , Europio/química , Ibuprofeno/análogos & derivados , Ibuprofeno/química , Cinética , Imagen por Resonancia Magnética , Ratones , Unión ProteicaRESUMEN
Four novel chelators (L1-L4) and their 89zirconium complexes were prepared and compared with the 89zirconium desferrioxamine B (DFO) complex. The new chelates are based on 1,4,7,10-tetraazacyclododecane (cyclen) and 1,4,8,11-tetraazacyclotetradecane (cyclam) scaffolds and present either three or four hydroxamate arms for coordination with Zr4+ ions with coordination numbers between six and eight. The 89Zr-L4 complex showed similar stability to that of 89Zr-DFO when incubated in either rat blood plasma or ethylenediaminetetraacetic acid challenge experiments. Positron imaging and biodistribution studies in mice showed that 89Zr-L4 had similar pharmacokinetic behavior to that of 89Zr-DFO, with rapid renal elimination and low residual activity in background tissues. A bifunctional version of L4 (L5) was synthesized and conjugated to trastuzumab; an anti-HER2/neu antibody. Immunopositron emission tomography imaging and biodistribution with 89Zr-L5-trastuzumab revealed high tumor to background ratios (tumor/blood ratio: 14.2 ± 2.25) and a high tumor specificity that was comparable to the performance of 89Zr-DFO-trastuzumab.
RESUMEN
An efficient synthesis of the C22-C40 domain of the azaspiracids is described. The synthetic route features a Nozaki-Hiyama-Kishi (NHK) coupling and chelation controlled Mukaiyama aldol reaction to access an acyclic intermediate and a double-intramolecular-hetero-Michael addition (DIHMA) to provide the FG-ring system bridged ketal.
Asunto(s)
Toxinas Marinas/síntesis química , Compuestos de Espiro/síntesis química , Toxinas Marinas/química , Estructura Molecular , Compuestos de Espiro/química , EstereoisomerismoRESUMEN
We report the synthesis and MR relevant properties of CyPic3A, a heptadentate chelator that forms ternary Gd(III) complexes of hydration state q = 2. [Gd(CyPic3A)(H2O)2](-) affords an r1 value of 5.70 mM(-1) s(-1) at 1.41 T and 310 K and displays thermodynamic stability and kinetic inertness comparable to FDA approved MR imaging probes.