RESUMO
Lysyl oxidase (LOX) is upregulated in highly stiff aggressive tumors, correlating with metastasis, resistance, and worse survival; however, there are currently no potent, safe, and orally bioavailable small molecule LOX inhibitors to treat these aggressive desmoplastic solid tumors in clinics. Here we discovered bi-thiazole derivatives as potent LOX inhibitors by robust screening of drug-like molecules combined with cell/recombinant protein-based assays. Structure-activity relationship analysis identified a potent lead compound (LXG6403) with â¼3.5-fold specificity for LOX compared to LOXL2 while not inhibiting LOXL1 with a competitive, time- and concentration-dependent irreversible mode of inhibition. LXG6403 shows favorable pharmacokinetic properties, globally changes ECM/collagen architecture, and reduces tumor stiffness. This leads to better drug penetration, inhibits FAK signaling, and induces ROS/DNA damage, G1 arrest, and apoptosis in chemoresistant triple-negative breast cancer (TNBC) cell lines, PDX organoids, and in vivo. Overall, our potent and tolerable bi-thiazole LOX inhibitor enhances chemoresponse in TNBC, the deadliest breast cancer subtype.
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Herein, DFT studies corroborating experimental results revealed that the shortest intramolecular hydrogen bonding distance of cis/trans-oxaphosphetane (OPA) oxygen with the CH-hydrogen of a triphenylphosphine phenyl ring provides good evidence for the attained olefin Z/E-selectivity in Wittig olefination of the studied examples. 2-Nitrobenzaldehyde, 3-nitrobenzaldehyde, 2-nitro-3-bromobenzaldehyde, 2-nitro-5-bromobenzaldehyde and 2-nitro-5-arylbenzaldehydes provided Z-nitrostilbenes with (2-chloro-4-hydroxy-3-methoxy-5-(methoxycarbonyl)benzyl) triphenylphosphonium chloride as the major products. However, 4-nitrobenzaldehyde and 2-nitro-6-bromobenzaldehydes furnished E-nitrostilbenes as the major products in high yields. Furthermore, the DFT computed intramolecular CH1/CH2-hydrogen bond distances with Cl/NO2 of selected stilbene derivatives were in good agreement with intramolecular hydrogen bond distances measured from single-crystal X-ray diffraction measurements.
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Serum tyrosine levels increase during aging, neurocognitive, metabolic, and cardiovascular disorders. However, calorie restriction (CR) and sleep lower serum tyrosine levels. We previously showed that tyrosine inhibits tyrosyl-tRNA synthetase (TyrRS)-mediated activation of poly-ADP-ribose polymerase 1 (PARP1). Here, we show that histone serine-ADP-ribosylation is decreased in Alzheimer's Disease (AD) brains, and increased tyrosine levels deplete TyrRS and cause neuronal DNA damage. However, dopamine and brain-derived neurotrophic factor (BDNF) increase TyrRS and histone serine-ADP-ribosylation. Furthermore, cis-resveratrol (cis-RSV) that binds to TyrRS mimicking a 'tyrosine-free' conformation increases TyrRS, facilitates histone serine-ADP-ribosylation-dependent DNA repair, and provides neuroprotection in a TyrRS-dependent manner. Conversely, trans-RSV that binds to TyrRS mimicking a 'tyrosine-like' conformation decreases TyrRS, inhibits serine-ADP-ribosylation-dependent DNA repair, and induces neurodegeneration in rat cortical neurons. Our findings suggest that age-associated increase in serum tyrosine levels may effect neurocognitive and metabolic disorders and offer a plausible explanation for divergent results obtained in clinical trials using resveratrol.
Assuntos
Histonas , Tirosina-tRNA Ligase , ADP-Ribosilação , Animais , Histonas/metabolismo , Ratos , Resveratrol/farmacologia , Serina/metabolismo , Tirosina/metabolismo , Tirosina-tRNA Ligase/metabolismoRESUMO
Unlike widely perceived, resveratrol (RSV) decreased the average lifespan and extended only the replicative lifespan in yeast. Similarly, although not widely discussed, RSV is also known to evoke neurite degeneration, kidney toxicity, atherosclerosis, premature senescence, and genotoxicity through yet unknown mechanisms. Nevertheless, in vivo animal models of diseases and human clinical trials demonstrate inconsistent protective and beneficial effects. Therefore, the mechanism of action of RSV that elicits beneficial effects remains an enigma. In a previously published work, we demonstrated structural similarities between RSV and tyrosine amino acid. RSV acts as a tyrosine antagonist and competes with it to bind to human tyrosyl-tRNA synthetase (TyrRS). Interestingly, although both isomers of RSV bind to TyrRS, only the cis-isomer evokes a unique structural change at the active site to promote its interaction with poly-ADP-ribose polymerase 1 (PARP1), a major determinant of cellular NAD+-dependent stress response. However, retention of trans-RSV in the active site of TyrRS mimics its tyrosine-bound conformation that inhibits the auto-poly-ADP-ribos(PAR)ylation of PARP1. Therefore, we proposed that cis-RSV-induced TyrRS-regulated auto-PARylation of PARP1 would contribute, at least in part, to the reported health benefits of RSV through the induction of protective stress response. This observation suggested that trans-RSV would inhibit TyrRS/PARP1-mediated protective stress response and would instead elicit an opposite effect compared to cis-RSV. Interestingly, most recent studies also confirmed the conversion of trans-RSV and its metabolites to cis-RSV in the physiological context. Therefore, the finding that cis-RSV and trans-RSV induce two distinct conformations of TyrRS with opposite effects on the auto-PARylation of PARP1 provides a potential molecular basis for the observed dichotomic effects of RSV under different experimental paradigms. However, the fact that natural RSV exists as a diastereomeric mixture of its cis and trans isomers and cis-RSV is also a physiologically relevant isoform has not yet gained much scientific attention.
Assuntos
Tirosina-tRNA Ligase , Animais , Dano ao DNA , Humanos , Poli(ADP-Ribose) Polimerase-1 , Resveratrol/farmacologiaRESUMO
Dopamine is critical for processing of reward and etiology of drug addiction. Astrocytes throughout the brain express dopamine receptors, but consequences of astrocytic dopamine receptor signaling are not well established. We found that extracellular dopamine triggered rapid concentration-dependent stellation of astrocytic processes that was not a result of dopamine oxidation but instead relied on both cAMP-dependent and cAMP-independent dopamine receptor signaling. This was accompanied by reduced duration and increased frequency of astrocytic Ca2+ transients, but little effect on astrocytic voltage-gated potassium channel currents. To isolate possible mechanisms underlying these structural and functional changes, we used whole-genome RNA sequencing and found prominent dopamine-induced enrichment of genes containing the CCCTC-binding factor (CTCF) motif, suggesting involvement of chromatin restructuring in the nucleus. CTCF binding to promoter sites bidirectionally regulates gene transcription and depends on activation of poly-ADP-ribose polymerase 1 (PARP1). Accordingly, antagonism of PARP1 occluded dopamine-induced changes, whereas a PARP1 agonist facilitated dopamine-induced changes on its own. These results indicate that astrocyte response to elevated dopamine involves PARP1-mediated CTCF genomic restructuring and concerted expression of gene networks. Our findings propose epigenetic regulation of chromatin landscape as a critical factor in the rapid astrocyte response to dopamine.SIGNIFICANCE STATEMENT Although dopamine is widely recognized for its role in modulating neuronal responses both in healthy and disease states, little is known about dopamine effects at non-neuronal cells in the brain. To address this gap, we performed whole-genome sequencing of astrocytes exposed to elevated extracellular dopamine and combined it with evaluation of effects on astrocyte morphology and function. We demonstrate a temporally dynamic pattern of genomic plasticity that triggers pronounced changes in astrocyte morphology and function. We further show that this plasticity depends on activation of genes sensitive to DNA-binding protein CTCF. Our results propose that a broad pattern of astrocyte responses to dopamine specifically relies on CTCF-dependent gene networks.
Assuntos
Astrócitos/efeitos dos fármacos , Astrócitos/ultraestrutura , Fator de Ligação a CCCTC/efeitos dos fármacos , Fator de Ligação a CCCTC/genética , Dopamina/farmacologia , Animais , Fator de Ligação a CCCTC/fisiologia , Sinalização do Cálcio/efeitos dos fármacos , Cromatina/genética , Cromatina/fisiologia , Fenômenos Eletrofisiológicos/fisiologia , Regulação da Expressão Gênica/efeitos dos fármacos , Genômica , Poli(ADP-Ribose) Polimerase-1/efeitos dos fármacos , Poli(ADP-Ribose) Polimerase-1/genética , Canais de Potássio de Abertura Dependente da Tensão da Membrana/efeitos dos fármacos , RNA/genética , Ratos , Ratos Sprague-Dawley , Análise de Sequência de RNA , Transcriptoma/efeitos dos fármacos , Transcriptoma/genéticaRESUMO
New conditions for dearomatizing samarium-ketyl (hetero)arene cyclizations are reported. In many examples of these samarium diiodide-mediated reactions, lithium bromide and water can be used as additives instead of the carcinogenic and mutagenic hexamethylphosphoramide (HMPA). The best results were obtained for the cyclizations of N-acylated indole derivatives delivering the expected indolines in good yields and excellent diastereoselectivities. A new type of cyclization delivering indolyl-substituted allene derivatives is also described. The scope and limitations of the lithium bromide/water system are discussed.
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Samarium diiodide mediated cyclizations of N-acylated indole derivatives bearing sulfinyl imine moieties afforded polycyclic tertiary carbinamines with moderate to excellent diastereoselectivities. Lithium bromide and water turned out to be the best additives to achieve these transformations in good yields. Using enantiopure sulfinyl imines the outcome strongly depends on the reactivity of the indole moiety. Whereas with unactivated indole derivatives desulfinylation and formation of racemic products was observed, indoles bearing electron-withdrawing substituents at C-3 afforded the polycyclic products with intact N-sulfinyl groups and with excellent diastereoselectivity, finally allowing the preparation of enantiopure tertiary carbinamines. The mechanisms of these processes are discussed.
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An unusual cascade rearrangement has been noticed as a competitive reaction during the treatment of norbornyl appended cyclopentanediols with a Lewis acid (LA): a BF3-Et2O mediated pinacol-pinacolone rearrangement. Deketalization and pinacolone rearrangement occur at two different sites in the molecule and are responsible for the observed cascade rearrangement product. However, deketalization appears to be triggering the cascade steps. The kinetically more stable pinacolone product with an exo-Me group was observed in the case of the bromo analogue, whereas, the thermodynamically more stable pinacolone product with an endo-Me group was observed in the case of the chloro analogue. Epimerization via tautomerization of one diastereomer to the other diastereomer under Lewis acid reflux conditions is possible. On the contrary, the diol equivalent epoxides provide only the diastereomeric mixture of pinacolone products under similar LA reaction conditions. The lower yields observed in the case of the epoxides are due to unwanted side reactions taking place between the two competitive reactive centers, namely, ketal and epoxide. Further, a sequence of elimination, nucleophilic substitution and Ritter type hydrolysis reactions of the epoxides resulted in unexpected elimination products. This transformation not only facilitates a regioselective epoxide opening, but also provides a new route for the preparation of allylic amides of the norbornyl appended cyclopentane ring system.
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The reaction of six substrates (diphenylacetylene, benzonitrile, methyl benzoate, phenylacetylene, naphthalene, and 1-chloro-4-ethylbenzene) with SmI(2) in the presence of MeOH or TFE was studied. The reactions were monitored under three different conditions: (a) irradiation, (b) irradiation in the presence of HMPA, and (c) reactions in the presence of HMPA in the dark. The combination of visible light and HMPA was found in some cases to be synergistic, in others to be additive, and in four cases to be inhibitive. The Marcus theory provides a good understanding of the synergistic and the additivity phenomena. The inhibitive effect is traced to the post electron transfer step in which Sm(3+) plays an important role. Once coordinated to HMPA, Sm(3+) is less capable of assisting in the protonation of the radical anion or the expulsion of the leaving group. Ranking according to the substrate's electron affinity shows that inhibition is manifested for the three least electrophilic substrates: phenylacetylene, naphthalene, and 1-chloro-4-ethylbenzene. Typical of these substrates is the short lifetime of their radical anions. Thus, if a step consecutive to electron transfer is slow and cannot compete successfully with the rapid back electron transfer, the benefit of having the electron transfer step enhanced is much reduced.
Assuntos
Hempa/química , Iodetos/química , Luz , Samário/química , Acetileno/análogos & derivados , Acetileno/química , Benzoatos/química , Estrutura Molecular , Naftalenos/química , Nitrilas/química , OxirreduçãoRESUMO
Despite their high electron-withdrawing strength, nitriles are not good electron acceptors and therefore are hard to reduce. In this work, using photostimulation in the visible region, we examined the reactivity of aliphatic and aromatic, mono- and dicyano compounds in reaction with SmI(2). A proton donor that complexes efficiently with SmI(2) must be used. Maximum yield was obtained at ca.0.2 M MeOH. Aromatic nitriles were more reactive than aliphatic nitriles, which exhibited negligible yields. Phenylacetonitrile presents an intermediate reactivity. The mechanism of the reaction involves coordination of the SmI(2) to the lone pair of the nitrile nitrogen followed by an inner sphere electron transfer. Surprisingly, m-dicyanobenzene was less reactive than the monocyano derivative benzonitrile. This was traced to the lower ability of the dicyano compound to coordinate to the SmI(2) due to, as was shown by quantum mechanical calculations, its lone pair having an energy significantly lower than that of benzonitrile. It is noteworthy that at the SmI(2) initial concentration used (0.04M), light penetrates only the 0.4 mm outer layer of the reaction mixture. Therefore the photostimulation effect observed was due to irradiation of only 4% of the total reaction volume, implying that under optimal conditions the effect should be 25 times larger.
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The reduction of azobenzene by SmI(2) in THF to give hydrazobenzene was investigated. The kinetics are first order in the substrate and first order in SmI(2). The kinetic order in MeOH is ca. 0.56, and in TFE it is ca. 0.2. The fractional order in the proton donors is interpreted as being a result of their acting in two opposing manners. In one the proton donor enhances the reaction by protonation of the radical anion, and in the other it slows the reaction by binding to the lone pair electrons of the nitrogen in the azobenzene. This hampers the fast inner-sphere electron-transfer mode. Experiments conducted in the presence of low concentrations of HMPA show rate enhancement suggesting that the SmI(2), which is partly coordinated to HMPA molecules, has some free sites to bind to the substrate. When more HMPA is added, it prevents the fast inner-sphere mechanism and the rate decreases. In this system, the increase in the reduction potential of SmI(2) caused by HMPA is similar to the rate enhancement by an inner sphere mechanism. In general, the replacement of a skeletal carbon by a nitrogen atom causes a significant rate enhancement.
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The reduction of the three imines, N-benzylidene aniline (BAI), N-benzylidenemethylamine (BMI), and benzophenone imine (BPI), with SmI(2) gives the reduced as well as coupled products. The reactions were found to be autocatalytic due to the formation of the trivalent samarium in the course of the reaction. When preprepared SmI(3) was added to the reaction mixture, the reaction rate increased significantly. However, the kinetics were found to be of zero order in SmI(2). This type of behavior is typical of surface catalysis with saturation of the catalytic sites. Although no solids are visible to the naked eye, the existence of microcrystals was proven by light microscopy as well as by dynamic light scattering analysis. Although HRTEM shows the existence of quantum dots in the solid, we were unable to make a direct connection between the existence of the quantum dots and the catalytic phenomenon. In the uncatalyzed reaction, the order of reactivity is BPI > BMI > BAI. This order does not conform to the electron affinity order of the substrates but rather to the nitrogen lone pair accessibility for complexation. This conclusion was further supported by using HMPA as a diagnostic probe for the existence of an inner sphere electron transfer reaction.