RESUMEN
Polyamines are positively charged alkylamines ubiquitous among eukaryotes, prokaryotes, and archaea. Humans obtain polyamines through dietary intake, metabolic production, or uptake of polyamines made by gut microbes. The polyamine biosynthetic pathway used by most gut microbes differs from that used by human cells. This alternative pathway employs carboxyspermidine dehydrogenase (CASDH), an enzyme with limited characterization. Here, we solved a 1.94 Å X-ray crystal structure of Bacteroides fragilis CASDH by molecular replacement. BfCASDH is composed of three domains with a fold similar to saccharopine dehydrogenase but with a distinct active site arrangement. Using steady-state methods, we determined kcat and kcat/Km for BfCASDH and Clostridium leptum CASDH using putrescine, diaminopropane, aspartate semi-aldehyde, NADH, and NADPH as substrates. These data revealed evidence of cooperativity in BfCASDH. Putrescine is the likely polyamine substrate and NADPH is the coenzyme used to complete the reaction, forming carboxyspermidine as a product. These data provide the first kinetic characterization of CASDH-a key enzyme in the production of microbial polyamines.
Asunto(s)
Bacteroides fragilis , Oxidorreductasas , Humanos , NADP , Oxidorreductasas/química , Oxidorreductasas/metabolismo , Poliaminas/metabolismo , Putrescina , Espermidina/metabolismo , Bacteroides fragilis/enzimologíaRESUMEN
The total synthesis and antileishmanial activity of deoxyalpinoid B is reported via a cationic gold-catalyzed Meyer-Schuster rearrangement. The activity of deoxyalpinoid B and a known inducer of oxidative stress, sulforaphane, against Leishmania donovani and Leishmania infantatum are both reported for the first time. Both compounds exhibit potent antileishmanial activity against both species. We hypothesize that the activation of intracellular oxidative stress is a key molecular response for the inhibition of Leishmania.
Asunto(s)
Antiprotozoarios , Leishmania donovani , Isotiocianatos/farmacología , Sulfóxidos/farmacologíaRESUMEN
The total syntheses of three structurally related natural products, deoxyalpinoid B, deoxyalpinoid A, and alpinoid F, are reported, and each features a Au(I)-catalyzed Meyer-Schuster rearrangement as the key step. The synthesis of alpinoid F is reported for the first time. The syntheses of these natural products, all of which exhibit potent anticancer activity, are readily amenable to the preparation of structural analogs.
RESUMEN
We report absolute integral cross section (ICS) measurements using a dual-source merged-fast-beams apparatus to study the titular reactions over the relative translational energy range of Er â¼ 0.01-10 eV. We used photodetachment of C- to produce a pure beam of atomic C in the ground electronic 3P term, with statistically populated fine-structure levels. The H2+ and D2+ were formed in an electron impact ionization source, with well known vibrational and rotational distributions. The experimental work is complemented by a theoretical study of the CH2+ electronic system in the reactant and product channels, which helps to clarify the possible reaction mechanisms underlying the ICS measurements. Our measurements provide evidence that the reactions are barrierless and exoergic. They also indicate the apparent absence of an intermolecular isotope effect, to within the total experimental uncertainties. Capture models, taking into account either the charge-induced dipole interaction potential or the combined charge-quadrupole and charge-induced dipole interaction potentials, produce reaction cross sections that lie a factor of â¼4 above the experimental results. Based on our theoretical study, we hypothesize that the reaction is most likely to proceed adiabatically through the 14A' and 14A'' states of CH2+via the reaction C(3P) + H2+(2Σ+g) â CH+(3Π) + H(2S). We also hypothesize that at low collision energies only H2+(v ≤ 2) and D2+(v ≤ 3) contribute to the titular reactions, due to the onset of dissociative charge transfer for higher vibrational v levels. Incorporating these assumptions into the capture models brings them into better agreement with the experimental results. Still, for energies ⪠0.1 eV where capture models are most relevant, the modified charge-induced dipole model yields reaction cross sections with an incorrect energy dependence and lying â¼10% below the experimental results. The capture cross section obtained from the combined charge-quadrupole and charge-induced dipole model better matches the measured energy dependence but lies â¼30-50% above the experimental results. These findings provide important guidance for future quasiclassical trajectory and quantum mechanical treatments of this reaction.
RESUMEN
The first total synthesis of the marine natural product avrainvilleol is reported. The total synthesis features the first application of the transition-metal-free coupling of a tosyl hydrazone and a boronic acid to the preparation of a complex natural product, and the first example of this coupling with a hindered diortho substituted hydrazone substrate.
Asunto(s)
Productos Biológicos/síntesis química , Glucolípidos/síntesis química , Productos Biológicos/química , Glucolípidos/química , Estructura MolecularRESUMEN
The first report of the antiviral activity of (+)-sattabacin against varicella-zoster virus (VZV) is described. Our results show that (+)-sattabacin potently inhibits the growth of VZV at concentrations in the range of other drugs commonly prescribed for VZV infection. Experiments detailing the synthesis of (+)-sattabacin, quantification of cytotoxicity and gene expression data in human fibroblast cells are also presented. Gene expression data was obtained through microarray analysis from human fibroblast cells exposed to sattabacin in order to identify a possible mechanism by which (+)-sattabacin inhibits VZV replication.
Asunto(s)
Antivirales/farmacología , Herpesvirus Humano 3/efectos de los fármacos , Hexanonas/farmacología , Antivirales/química , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Hexanonas/síntesis química , Hexanonas/química , Humanos , Estructura MolecularRESUMEN
Purpose: High-dose-rate electronic brachytherapy (eBx) is a non-surgical treatment option for non-melanoma skin cancer (NMSC) patients. This study assessed long-term effectiveness and safety of eBx for the treatment of NMSC. Material and methods: A chart review was conducted to identify subjects who had five or more years since their last eBx treatment fraction. Subjects meeting these criteria were contacted to determine their interest in participating in a long-term follow-up study. Those who agreed, underwent a follow-up visit where consent was obtained, and their lesions were clinically assessed for recurrence and long-term skin toxicities. History and demographic data were retrospectively collected, and treatment method was verified. Results: 183 subjects with 185 lesions were enrolled into this study at four dermatology centers in two practices in California. Three subjects in the analysis were less than 5 years from the last treatment to follow-up visit. All lesions were stage 1 basal cell carcinoma, squamous cell carcinoma, or squamous cell carcinoma in situ. Recurrence rate for the 183 subjects was 1.1%. Long-term skin toxicities were reported in 70.0% of the subjects. Hypopigmentation grade 1 was observed in 65.9% of the lesions, telangiectasia grade 1 was seen in 22.2%, scarring grade 1 in two subjects (1.1%), hyperpigmentation grade 1 in two subjects (1.1%), and induration grade 2 in one patient (0.5%). The induration grade 2 was located on the upper back and did not limit instrumental activities of daily living (ADLs). Conclusions: Electronic brachytherapy for the treatment of non-melanoma skin cancer is safe and effective, showing excellent long-term 98.9% local control through a median follow-up of 7.6 years (n = 183), with minimal long-term toxicities.
RESUMEN
Stephacidin and notoamide natural products belong to a group of prenylated indole alkaloids containing a core bicyclo[2.2.2]diazaoctane ring system. These bioactive fungal secondary metabolites have a range of unusual structural and stereochemical features but their biosynthesis has remained uncharacterized. Herein, we report the first biosynthetic gene cluster for this class of fungal alkaloids based on whole genome sequencing of a marine-derived Aspergillus sp. Two central pathway enzymes catalyzing both normal and reverse prenyltransfer reactions were characterized in detail. Our results establish the early steps for creation of the prenylated indole alkaloid structure and suggest a scheme for the biosynthesis of stephacidin and notoamide metabolites. The work provides the first genetic and biochemical insights for understanding the structural diversity of this important family of fungal alkaloids.
Asunto(s)
Antineoplásicos/metabolismo , Aspergillus/genética , Factores Biológicos/biosíntesis , Genoma , Alcaloides Indólicos/metabolismo , Animales , Antineoplásicos/química , Aspergillus/metabolismo , Factores Biológicos/química , Factores Biológicos/metabolismo , Alcaloides Indólicos/química , Conformación Molecular , Prenilación , EstereoisomerismoRESUMEN
The bicyclo[2.2.2]diazaoctane ring system is common to a number of highly biologically active secondary metabolites isolated from numerous species of fungi. In this tutorial review, we describe the varied synthetic approaches that have been employed to construct this ring system in the course of recent total synthesis endeavors, and this review should be of interest to synthetic organic chemists and natural product chemists. Detailed herein are a number of synthetic disconnections including intramolecular S(N)2' cyclizations, biomimetic Diels-Alder reactions, radical cyclizations, and cationic cascade reactions.
Asunto(s)
Compuestos Bicíclicos con Puentes/síntesis química , Compuestos de Diazonio/síntesis química , Octanos/síntesis química , Alcaloides Indólicos , Indolizinas , Prenilación , Compuestos de EspiroRESUMEN
The preparation and biological activity of various structural analogs of the malbrancheamides are disclosed. The impact of indole chlorination, C-12a relative stereochemistry, and bicyclo[2.2.2]diazaoctane core oxidation state on the ability of these analogs to inhibit calmodulin dependent phosphodiesterase (PDE1) was studied, and a number of potent compounds were identified.
Asunto(s)
Calmodulina/antagonistas & inhibidores , Calmodulina/química , Alcaloides Indólicos/síntesis química , Alcaloides Indólicos/farmacología , Alcaloides/química , Cloro/química , Clorpromazina/química , Fosfodiesterasas de Nucleótidos Cíclicos Tipo 1/química , Diseño de Fármacos , Humanos , Alcaloides Indólicos/química , Indoles/química , Concentración 50 Inhibidora , Modelos Químicos , Oxígeno/química , EstereoisomerismoRESUMEN
The first application of the Pauson-Khand reaction (PKR) to the synthesis of azabridged bicyclic structures is described. Compounds containing azabicyclo[3.3.1]nonane and azabicyclo[3.2.1]octane rings fused to cyclopentenones were efficiently constructed via the PKR of cis-2,6-disubstituted N-acyl piperidine enyne substrates, many of which can be readily prepared from 4-methoxypyridine in a few steps. Moreover, the PKR of cis-2,6-disubstituted piperazine enynes allowed the preparation of diazabicyclo[3.3.1]nonanes fused to cyclopentenones. This new strategy for the synthesis of azabridged bicyclic frameworks was exploited as a key step in a concise, enantioselective total synthesis of the macroline alklaoid (-)-alstonerine.
RESUMEN
A novel enantioselective total synthesis of (-)-alstonerine has been completed that requires only 15 steps from L-tryptophan. The synthesis features the first application of a Pauson-Khand reaction to synthesize an azabridged bicyclic skeleton. [reaction: see text]
Asunto(s)
Alcaloides/síntesis química , Alcaloides Indólicos/síntesis química , Compuestos Aza/química , Compuestos Bicíclicos con Puentes/química , Modelos Químicos , Estereoisomerismo , Triptófano/químicaRESUMEN
[reaction: see text] Three novel domino reaction processes have been discovered and developed that employ the regioselective and stereoselective [Rh(CO)(2)Cl](2)-catalyzed alkylations of allylic trifluoroacetates with alpha-substituted sodiomalonates followed by an intramolecular Pauson-Khand annulation, a [5 + 2] cycloaddition, or a cycloisomerization. A unique aspect of the methodology is that a single catalyst is used to effect sequential transformations simply by increasing the temperature for the second reaction.
Asunto(s)
Compuestos Alílicos/química , Rodio/química , Compuestos Alílicos/síntesis química , Catálisis , Ciclización , Estructura Molecular , EstereoisomerismoRESUMEN
[Rh(CO)(2)Cl](2) has been found to possess the unusual property of catalyzing allylic alkylations of unsymmetrical allylic carbonates with high levels of regioselectivity to provide products arising from substitution at the carbon atom bearing the leaving group, irrespective of the structure of the starting carbonate. The substitution reaction occurs with retention of stereochemistry at the reacting center, and the carbon-carbon double-bond stereochemistry of primary (Z)-allylic carbonates is maintained. [reaction: see text]
Asunto(s)
Compuestos Alílicos/química , Cloruros/química , Rodio/química , Alquilación , Carbonatos/química , Catálisis , Estructura Molecular , EstereoisomerismoRESUMEN
The concept of targeting cancer therapeutics toward specific mutations or abnormalities in tumor cells, which are not found in normal tissues, has the potential advantages of high selectivity for the tumor and correspondingly low secondary toxicities. Many human malignancies display activating mutations in the Ras family of signal-transducing genes or over-activity of p21(Ras)-signaling pathways. Carcinoid and other neuroendocrine tumors have been similarly demonstrated to have activation of Ras signaling directly by mutations in Ras, indirectly by loss of Ras-regulatory proteins, or via constitutive activation of upstream or downstream effector pathways of Ras, such as growth factor receptors or PI(3)-kinase and Raf/mitogen-activated protein kinases. We previously reported that aberrant activation of Ras signaling sensitizes cells to apoptosis when the activity of the PKCδ isozyme is suppressed and that PKCδ suppression is not toxic to cells with normal levels of p21(Ras) signaling. We demonstrate here that inhibition of PKCδ by a number of independent means, including genetic mechanisms (shRNA) or small-molecule inhibitors, is able to efficiently and selectively repress the growth of human neuroendocrine cell lines derived from bronchopulmonary, foregut, or hindgut tumors. PKCδ inhibition in these tumors also efficiently induced apoptosis. Exposure to small-molecule inhibitors of PKCδ over a period of 24 âh is sufficient to significantly suppress cell growth and clonogenic capacity of these tumor cell lines. Neuroendocrine tumors are typically refractory to conventional therapeutic approaches. This Ras-targeted therapeutic approach, mediated through PKCδ suppression, which selectively takes advantage of the very oncogenic mutations that contribute to the malignancy of the tumor, may hold potential as a novel therapeutic modality.
Asunto(s)
Proliferación Celular , Tumores Neuroendocrinos/terapia , Proteína Quinasa C-delta/antagonistas & inhibidores , Acetofenonas/farmacología , Animales , Apoptosis , Benzopiranos/farmacología , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Neoplasias Gastrointestinales , Técnicas de Silenciamiento del Gen , Humanos , Neoplasias Pulmonares , Ratones , Células 3T3 NIH , Tumores Neuroendocrinos/enzimología , Tumores Neuroendocrinos/genética , Proteína Quinasa C-delta/genética , Proteína Quinasa C-delta/metabolismoRESUMEN
The Diels-Alder reaction is one of the most well-studied, synthetically useful organic transformations. While a significant number of naturally occurring substances are postulated to arise by biosynthetic Diels-Alder reactions, rigorous confirmation of a mechanistically distinct natural Diels-Alderase enzyme remains elusive. Within this context, several related fungi within the Aspergillus genus produce a number of metabolites of opposite absolute configuration including (+)- or (-)-versicolamide B. These alkaloids are hypothesized to arise via biosynthetic Diels-Alder reactions implying that each Aspergillus species possesses enantiomerically distinct Diels-Alderases. Herein, experimental validation of these biosynthetic proposals via deployment of the IMDA reaction as a key step in the asymmetric total syntheses of (+)- and (-)-versicolamide B is described. Laboratory validation of the proposed biosynthetic Diels-Alder construction, coupled with the secondary metabolite profile of the producing fungi, reveals that each Aspergillus species has evolved enantiomerically distinct indole oxidases, as well as enantiomerically distinct Diels-Alderases.
Asunto(s)
Alcaloides Indólicos/metabolismo , Aspergillus/enzimología , Aspergillus/metabolismo , Dicroismo Circular , Ciclización , Dioxigenasas/metabolismo , Proteínas Fúngicas/metabolismo , Alcaloides Indólicos/química , EstereoisomerismoRESUMEN
An advanced metabolite, named premalbrancheamide, involved in the biosynthesis of malbrancheamide (1) and malbrancheamide B (2) has been synthesized in double (13)C-labeled form and was incorporated into the indole alkaloid 2 by Malbranchea aurantiaca. In addition, premalbrancheamide has been detected as a natural metabolite in cultures of M. aurantiaca. The biosynthetic implications of these experiments are discussed.
Asunto(s)
Amidas/química , Amidas/metabolismo , Familia de Multigenes/genética , Onygenales/química , Onygenales/metabolismo , Marcaje Isotópico , Espectrometría de Masas , Estructura Molecular , Onygenales/genéticaRESUMEN
The biomimetic total syntheses of both malbrancheamide and malbrancheamide B are reported. The synthesis of the two monochloro species enabled the structure of malbrancheamide B to be unambiguously assigned. The syntheses each feature an intramolecular Diels-Alder reaction of a 5-hydroxypyrazin-2(1H)-one to construct the bicyclo[2.2.2]diazaoctane core, which has also been proposed as the biosynthetic route to these compounds.
Asunto(s)
Biomimética , Compuestos Heterocíclicos de 4 o más Anillos/síntesis química , Amidas/química , Compuestos Heterocíclicos de 4 o más Anillos/química , Alcaloides Indólicos , Estructura Molecular , Oxidación-ReducciónRESUMEN
A novel regio- and stereoselective [Rh(CO)2Cl]2-catalyzed allylic alkylation of unsymmetrical allylic carbonates was discovered. The regioselectivity of the reaction favors product ratios in which substitution occurs at the carbon bearing the leaving group. When an enantiomerically enriched carbonate (> or = 99% ee) was examined, the Rh(I)-catalyzed allylic alkylation proceeded stereoselectively to provide the alkylation product with retention of absolute stereochemistry (98% ee). To establish the scope of the [Rh(CO)2Cl]2-catalyzed allylic alkylation, a variety of carbon and heteroatom nucleophiles were examined and the results described. As an application of the Rh(I)-catalyzed allylic alkylation, a series of novel domino reactions have been developed that couple the unique regio- and stereoselective [Rh(CO)2Cl]2-catalyzed alkylation of allylic trifluoroacetates with an intramolecular Pauson-Khand annulation, a cycloisomerization, or a [5+2] cycloaddition. A unique aspect of the method described is the use of a single catalyst to effect sequential transformations in which the catalytic activity is moderated simply by controlling the reaction temperature. Implementation of such processes provides a rapid and efficient entry to a variety of bicyclic carbon skeletons from simple precursors.