RESUMEN
Conventional solid oral dosage form development is not typically challenged by reliance on an amorphous drug substance as a direct ingredient in the drug product, as this may result in product development hurdles arising from process design and scale-up, control of physical quality attributes, drug product processability and stability. Here, we present the Chemistry, Manufacturing and Controls development journey behind the successful commercialization of an amorphous drug substance, Elagolix Sodium, a first-in-class, orally active gonadotropin-releasing hormone antagonist. The reason behind the lack of crystalline state was assessed via Molecular Dynamics (MD) at the molecular and inter-molecular level, revealing barriers for nucleation due to prevalence of intra-molecular hydrogen bond, repulsive interactions between active pharmaceutical ingredient (API) molecules and strong solvation effects. To provide a foundational basis for the design of the API manufacturing process, we modeled the solvent-induced plasticization behavior experimentally and computationally via MD for insights into molecular mobility. In addition, we applied material science tetrahedron concepts to link API porosity to drug product tablet compressibility. Finally, we designed the API isolation process, incorporating computational fluid dynamics modeling in the design of an impinging jet mixer for precipitation and solvent-dependent glass transition relationships in the cake wash, blow-down and drying process, to enable the consistent manufacture of a porous, non-sintered amorphous API powder that is suitable for robust drug product manufacturing.
Asunto(s)
Simulación de Dinámica Molecular , Pirimidinas , Comprimidos , Administración Oral , Pirimidinas/química , Pirimidinas/administración & dosificación , Composición de Medicamentos/métodos , Cristalización , Química Farmacéutica/métodos , Porosidad , Enlace de Hidrógeno , Estabilidad de Medicamentos , Hidrocarburos FluoradosRESUMEN
A 2-phenyl-3-difluoromethoxy-pyridinyl moiety features in potent phosphodiesterase 4D inhibitors that are considered to be candidate radiotracers for positron emission tomography if they are labeled with fluorine-18. Fluorine-18 could be installed as desired at the 3'-phenyl position with acridinium-mediated photoredox radiodeoxyfluorination in homologues bearing variously substituted 3'-aryloxy groups. However, a distal 3-difluoromethoxide (-OCHF2) group strongly competes as a leaving group, especially when an electron-deficient aryloxy group is present at position 3'. A yield of up to 50% may occur without observable 19F for 18F exchange.
Asunto(s)
Radioisótopos de Flúor , Oxidación-Reducción , Piridinas , Piridinas/química , Piridinas/síntesis química , Radioisótopos de Flúor/química , Estructura Molecular , Procesos Fotoquímicos , Halogenación , Hidrocarburos Fluorados/química , Hidrocarburos Fluorados/síntesis químicaRESUMEN
Nucleic acid duplexes are typically analyzed in non-denaturing conditions. Melting temperature (Tm) is the property used to measure duplex stability; however, it is not known how the chromatographic conditions and mobile phase composition affect the duplex stability. We employed differential scanning calorimetry (DSC) method to measure the melting temperature of chemically modified silencing RNA duplex (21 base pairs, 0.15 mM duplex concentration) in mobile phases commonly used in reversed-phase, ion-pair reversed-phase, size exclusion and hydrophilic interaction chromatography. We investigated mobile phases consisting of ammonium acetate, alkylammonium ion-pairing reagents, alkali-ion chlorides, magnesium chloride, and additives including methanol, ethanol, acetonitrile and hexafluoroisopropanol. Increasing buffer concentration enhanced the duplex stability (Tm was 67.1 - 78.2 °C for 10-100 mM [Na+] concentration). The melting temperature decreases with the increase in cation size (70.2 °C in 10 mM [Li+], 68.1 °C in 10 mM [NH4+], 65.6 °C in 10 mM [Cs+], and 56.6 °C in 10 mM [triethylammonium+] solutions). Inclusion of 20 % of organic solvent in buffer reduced the melting temperature by 1-3 °C, and denaturation power increases in the order MeOHAsunto(s)
Rastreo Diferencial de Calorimetría
, ARN Interferente Pequeño
, ARN Interferente Pequeño/química
, Estabilidad del ARN
, Cromatografía de Fase Inversa/métodos
, Acetonitrilos/química
, Acetatos/química
, Metanol/química
, Interacciones Hidrofóbicas e Hidrofílicas
, Solventes/química
, Propanoles/química
, Cromatografía Liquida/métodos
, Etanol/química
, Temperatura de Transición
, Cromatografía en Gel/métodos
, Cloruro de Magnesio/química
, Hidrocarburos Fluorados
RESUMEN
TMEM16A, a member of the Transmembrane protein 16 family, serves as the molecular basis for calcium activated chloride channels (CaCCs). We use RT-PCR to demonstrate the expression of TMEM16A in the neurons of Helicoverpa armigera, and record the CaCCs current of acute isolated neurons of H. armigera for the first time using patch clamp technology. In order to screen effective inhibitors of calcium-activated chloride channels, the inhibitory effects of four chloride channel inhibitors, CaCCinh-A01, NPPB, DIDS, and SITS, on CaCCs were compared. The inhibitory effects of the four inhibitors on the outward current of CaCCs were CaCCinh-A01 (10 µM, 56.31 %), NPPB (200 µM, 43.69 %), SITS (1 mM, 12.41 %) and DIDS (1 mM, 13.29 %). Among these inhibitors, CaCCinh-A01 demonstrated the highest efficacy as a blocker. To further explore whether calcium channel proteins can serve as potential targets of pyrethroids, we compared the effects of (type I) tefluthrin and (type II) deltamethrin on CaCCs. 10 µM and 100 µM tefluthrin can stimulate a large tail current in CaCCs, prolonging their deactivation time by 10.44 ms and 31.49 ms, and the V0.5 shifted in the hyperpolarization by 2-8 mV. Then, deltamethrin had no obvious effect on the deactivation and activation of CaCCs. Therefore, CaCCs of H. armigera can be used as a potential target of pyrethroids, but type I and type II pyrethroids have different effects on CaCCs.
Asunto(s)
Canales de Cloruro , Insecticidas , Mariposas Nocturnas , Neuronas , Piretrinas , Animales , Insecticidas/toxicidad , Insecticidas/farmacología , Piretrinas/toxicidad , Piretrinas/farmacología , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Canales de Cloruro/metabolismo , Canales de Cloruro/antagonistas & inhibidores , Mariposas Nocturnas/efectos de los fármacos , Anoctamina-1/metabolismo , Anoctamina-1/antagonistas & inhibidores , Proteínas de Insectos/metabolismo , Proteínas de Insectos/antagonistas & inhibidores , Proteínas de Insectos/genética , Potenciales de la Membrana/efectos de los fármacos , Técnicas de Placa-Clamp , Nitrobenzoatos/farmacología , Helicoverpa armigera , Ciclopropanos , Hidrocarburos FluoradosRESUMEN
BACKGROUND: Elagolix, an approved non-peptide GnRH antagonist, shows promise in relieving endometriosis-related pain, but its short- and mid-term efficacy and potential side effects are still under investigation. OBJECTIVE: The aim is to provide data for therapeutic applications by methodically evaluating elagolix's safety and effectiveness in treating endometriosis-related pain. METHODS: Databases such as PubMed, Embase, Cochrane Library, Web of Science, ClinicalTrials.gov, and others were thoroughly searched. The search time was from the establishment date to September 2023. The study included randomized controlled trials (RCTs) that compared the efficacy of elagolix versus placebo in treating endometriosis-associated pain. After data extraction and literature scanning, quality assessment was carried out using Quality evaluation was carried out using the bias risk assessment tool suggested by the Cochrane Reviewers' Handbook 5.1.0 after literature screening and data extraction. Stata 15.0 was used to do the meta-analysis. RESULTS: In total, five RCTs involving 2056 patients were included in the analysis. The meta-analysis demonstrated a significant superiority of elagolix over placebo in the management of endometriosis-related pain, specifically in endometriosis pain [WMD=-0.77, 95% CI (-1.00, -0.53), P<0.001], as well as in non-menstrual pelvic pain, daily assessment of dysmenorrhea (DYS), and dyspareunia (DYSP), all of which are associated with endometriosis. Regarding safety, no discernible variation was observed in the incidence of serious adverse responses between the elagolix and placebo groups [RR=0.90, 95% CI (0.58, 1.40), P=0.643]. Conversely, the elagolix group exhibited a significantly higher incidence rate of general adverse responses [RR = 1.34, 95% CI (1.18, 1.52), P<0.001] compared to the control group. CONCLUSIONS: The efficacy of elagolix in reducing pain in premenopausal women with endometriosis has been demonstrated over the short- to mid-term. However, careful monitoring for potential adverse effects is essential throughout the treatment duration.
Asunto(s)
Endometriosis , Hidrocarburos Fluorados , Pirimidinas , Humanos , Endometriosis/complicaciones , Endometriosis/tratamiento farmacológico , Femenino , Hidrocarburos Fluorados/uso terapéutico , Pirimidinas/uso terapéutico , Pirimidinas/efectos adversos , Dolor Pélvico/tratamiento farmacológico , Dolor Pélvico/etiología , Ensayos Clínicos Controlados Aleatorios como Asunto , Resultado del Tratamiento , Dispareunia/tratamiento farmacológico , Dispareunia/etiologíaRESUMEN
Residues of pesticides in milk may pose a threat to human health. This study aimed to develop a liquid-phase microextraction (LPME) method using hexafluoroisopropanol (HFIP)-based supramolecular solvent (SUPRAS) for the simultaneous extraction and purification of four pesticides (boscalid, novaluron, cypermethrin and bifenthrin) in milk. Pesticides were extracted using SUPRAS prepared with nonanol and HFIP, and the extraction efficiency was analyzed. Results showed satisfactory recoveries ranging from 80.8%-111.0%, with relative standard deviations (RSDs) of <6.4%. Additionally, satisfactory linearities were observed, with correlation coefficients >0.9952. The limits of quantification (LOQs) were in the range of 1.8 µg·L-1-14.0 µg·L-1. The established method demonstrated high extraction efficiency with a short operation time (15 mins) and low solvent consumption (2.7 mL). The HFIP-based SUPRAS LPME method offers a convenient and efficient approach for the extraction of pesticides from milk, presenting a promising alternative to conventional techniques.
Asunto(s)
Contaminación de Alimentos , Microextracción en Fase Líquida , Leche , Solventes , Microextracción en Fase Líquida/métodos , Leche/química , Animales , Solventes/química , Contaminación de Alimentos/análisis , Residuos de Plaguicidas/aislamiento & purificación , Residuos de Plaguicidas/química , Residuos de Plaguicidas/análisis , Hexanoles/química , Bovinos , Plaguicidas/aislamiento & purificación , Plaguicidas/química , Plaguicidas/análisis , Hidrocarburos Fluorados , PropanolesRESUMEN
There is an unmet clinical need for effective anticoagulant therapies for the management of thromboembolic diseases that are not associated with a relevant risk of bleeding. Asundexian (BAY 2433334) is an oral, direct, small-molecule inhibitor of activated factor XI (FXIa). Phase I data from healthy Caucasian male participants indicated predictable pharmacokinetic (PK) and pharmacodynamic (PD) profiles and no clinically relevant bleeding-related adverse events (AEs). Reported here are data from two phase I, randomized, placebo-controlled, single- and multiple-dose escalation studies of asundexian conducted in 60 healthy men: 24 Japanese and 36 Chinese. Baseline characteristics were comparable between the treatment groups. All treatment-emergent AEs were mild, with no serious AEs or AEs of special interest reported. Systemic exposure to asundexian increased dose proportionally after single or multiple dosing, with relatively low accumulation following multiple once-daily dosing in both Chinese and Japanese volunteers. Asundexian induced dose-dependent prolongation of activated partial thromboplastin time and inhibition of FXIa activity, with no effects on prothrombin time or FXI concentration in Japanese participants. There were no clinically relevant interethnic differences in PK profile across the Japanese, Chinese, and Caucasian (data from the previous phase I study) participants and no clinically relevant difference in PD response between Japanese and Caucasian participants.
Asunto(s)
Voluntarios Sanos , Población Blanca , Humanos , Masculino , Adulto , Adulto Joven , Relación Dosis-Respuesta a Droga , Anticoagulantes/farmacocinética , Anticoagulantes/administración & dosificación , Anticoagulantes/efectos adversos , Persona de Mediana Edad , Método Doble Ciego , Tiempo de Tromboplastina Parcial , Factor XIa/antagonistas & inhibidores , Administración Oral , Coagulación Sanguínea/efectos de los fármacos , Pueblos del Este de Asia , Benzamidas , Hidrocarburos Fluorados , TriazolesRESUMEN
Populations of various economic species of wireworms are increasing in the key cereal crop production areas of Canada and the United States. To address this problem, seed treatments are under development that both provide crop protection and significantly reduce populations equivalent in effectiveness to the formerly used but now deregistered organochlorine lindane. Herein, we evaluated isocycloseram (PLINAZOLIN technology), the first isoxazoline (GABA-gated Chloride Channel Allosteric Modulator) agricultural insecticide, as a seed treatment for the protection of cereal crops from the sugarbeet wireworm, Limonius californicus (Mannerheim). In wheat and barley field trials conducted over 4 years under extreme wireworm pressure, isocycloseram applied as a seed treatment at 5.0-7.5 g AI/100 kg seed was as effective as or more effective than the current industry standard thiamethoxam at 20.0 g AI/100 kg seed in protecting crop stand and yield. Isocycloseram also reduced neonate wireworms (produced from eggs during the growing season) and resident wireworms (in the field at the time of planting) to levels expected from the formerly used seed treatment lindane.
Asunto(s)
Escarabajos , Hordeum , Insecticidas , Semillas , Triticum , Animales , Escarabajos/efectos de los fármacos , Control de Insectos , Larva/crecimiento & desarrollo , Protección de Cultivos/métodos , Ciclopropanos , Hidrocarburos FluoradosRESUMEN
Enzymes capable of assimilating fluorinated feedstocks are scarce. This situation poses a challenge for the biosynthesis of fluorinated compounds used in pharmaceuticals, agrochemicals, and materials. We developed a photoenzymatic hydrofluoroalkylation that integrates fluorinated motifs into olefins. The photoinduced promiscuity of flavin-dependent ene-reductases enables the generation of carbon-centered radicals from iodinated fluoroalkanes, which are directed by the photoenzyme to engage enantioselectively with olefins. This approach facilitates stereocontrol through interaction between a singular fluorinated unit and the enzyme, securing high enantioselectivity at ß, γ, or δ positions of fluorinated groups through enzymatic hydrogen atom transfer-a process that is notably challenging with conventional chemocatalysis. This work advances enzymatic strategies for integrating fluorinated chemical feedstocks and opens avenues for asymmetric synthesis of fluorinated compounds.
Asunto(s)
Alquenos , Halogenación , Hidrocarburos Fluorados , Oxidorreductasas , Procesos Fotoquímicos , Alquenos/química , Alquilación , Hidrocarburos Fluorados/química , Oxidorreductasas/química , Estereoisomerismo , CatálisisRESUMEN
The potential ecological risk of per- and polyfluorinated alkyl substances (PFASs) in phytoremediation has raised social concerns, promoting a need to better understand their distribution and risks in the recovery process of aquatic plants. Herein, we aim to fill this knowledge gap by investigating the distribution and ecotoxicological effects of PFASs on the structure and function of water-macrophyte-sediment microcosm systems. Among the entire system, 63.0 %-73.1 % PFOA was found in sediments and submerged plants, however, 52.5 %-53.0 % of PFPeA and 47.0 %-47.5 % of PFBS remained in the water under different treatments. PFOA was more bioavailable than the other substances, as demonstrated by the bioaccumulation factors (BAF) with ranges exposed to PFPeA and PFBS. Bioaccumulation PFASs induced plant oxidative stress which generates enzymes to suppress superoxide, and disturbed the processes of lysine biosynthesis, in which allysine, meso-2,6-diaminoheptanedioate, and Nsuccinyl-2-amino-6-ketopimelate were downregulated. PFASs were detected in the propagator (turions) of an ecological restoration species, where short-chain PFASs (70.1 % and 45.7 % for 2 or 20 µg/L PFAS exposure, respectively) were found to spread further into new individuals and profoundly influence ecological processes shaping populations. PFASs significantly enhanced the number of microbial species in the sediment, but the degree of differentiation in the microbial community structure was not significantly different. This study enhances our understanding of the ecological mechanisms of PFASs in the water-macrophyte-sediment systems and potential threats to the recovery process of macrophytes.
Asunto(s)
Biodegradación Ambiental , Contaminantes Químicos del Agua , Contaminantes Químicos del Agua/análisis , Fluorocarburos/metabolismo , Plantas/metabolismo , Plantas/efectos de los fármacos , Hidrocarburos Fluorados/metabolismo , Sedimentos Geológicos/químicaRESUMEN
Oxathiapiprolin (OXA), which targets the oxysterol-binding protein (OSBP), is an outstanding piperidinyl thiazole isoxazoline (PTI) fungicide that can be used to control oomycetes diseases. In this study, starting from the structure of OXA, a series of novel OSBP inhibitors were designed and synthesized by introducing an indole moiety to replace the pyrazole in OXA. Finally, compound b24 was found to exhibit the highest control effect (82%) against cucumber downy mildew (CDM) in the greenhouse at a very low dosage of 0.069 mg/L, which was comparable to that of OXA (88%). Furthermore, it showed better activity against potato late blight (PLB) than other derivatives of indole. The computational results showed that the R-conformation of b24 should be the dominant conformation binding to PcOSBP. The results of the present work indicate that the 3-fluorine-indole ring is a favorable fragment to increasing the electronic energy when binding with PcOSBP. Furthermore, compound b24 could be used as a lead compound for the discovery of new OSBP inhibitors.
Asunto(s)
Fungicidas Industriales , Enfermedades de las Plantas , Fungicidas Industriales/química , Fungicidas Industriales/farmacología , Enfermedades de las Plantas/microbiología , Relación Estructura-Actividad , Indoles/química , Indoles/farmacología , Cucumis sativus/química , Cucumis sativus/microbiología , Oomicetos/efectos de los fármacos , Solanum tuberosum/química , Estructura Molecular , Simulación del Acoplamiento Molecular , Descubrimiento de Drogas , Hidrocarburos Fluorados , PirazolesRESUMEN
In our continuing effort devoted at developing agents targeting the EphA2 receptor by means of protein-protein interaction (PPI) inhibitors, we report here the design and synthesis of a new class of l-ß-homotryptophan conjugates of 3-ß-hydroxy-Δ5-cholenic acid bearing a set of arylsulfonyl substituents at the indole nitrogen atom. An extensive structure-activity relationship (SAR) analysis indicates that the presence of a bulky lipophilic moiety at the indole nitrogen is fundamental for improving potency on the EphA2 receptor, while abrogating activity on the EphB1-EphB3 receptor subtypes. A rational exploration, guided by the combined application of an experimental design on σp and π physicochemical descriptors and docking simulations, led to the discovery of UniPR1454, a 1-(4-(trifluoromethyl)phenyl)sulfonyl derivative acting as potent and competitive EphA2 antagonist able to inhibit ephrin-A1 dependent signals and to reduce proliferation of glioblastoma (U251) cell line at micromolar concentration.
Asunto(s)
Antineoplásicos , Proliferación Celular , Descubrimiento de Drogas , Ensayos de Selección de Medicamentos Antitumorales , Glioblastoma , Indoles , Receptor EphA2 , Humanos , Antineoplásicos/farmacología , Antineoplásicos/síntesis química , Antineoplásicos/química , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Glioblastoma/tratamiento farmacológico , Glioblastoma/patología , Glioblastoma/metabolismo , Indoles/farmacología , Indoles/química , Indoles/síntesis química , Simulación del Acoplamiento Molecular , Estructura Molecular , Receptor EphA2/antagonistas & inhibidores , Receptor EphA2/metabolismo , Relación Estructura-Actividad , Hidrocarburos Fluorados/síntesis química , Hidrocarburos Fluorados/química , Hidrocarburos Fluorados/farmacologíaRESUMEN
Continuous growth in fluoroarene production has led to environmental pollution and health concerns owing to their persistence, which is attributed to the stable C-F bond in their structures. Herein, we investigated fluoroarene decomposition via hydrodefluorination using a rhodium-based catalyst, focusing on the effects of the chemical structure and functional group on the defluorination yield. Most compounds, except (pentafluoroethyl)benzene, exhibited full or partial reduction with pseudo-first-order rate constants in the range of 0.002-0.396 min-1 and defluorination yields of 0%-100%. Fluoroarenes with hydroxyl, methyl, and carboxylate groups were selected to elucidate how hydrocarbon and oxygen-containing functional groups influence the reaction rate and defluorination. Inhibition of the reaction rate and defluorination yield based on functional groups increased in the order of hydroxyl < methyl < carboxylate, which was identical to the order of the electron-withdrawing effect. Fluoroarenes with polyfluoro groups were also assessed; polyfluoro groups demonstrated a different influence on catalyst activity than non-fluorine functional groups because of fluorine atoms in the substituents undergoing defluorination. The reaction kinetics of (difluoromethyl)fluorobenzenes and their intermediates suggested that hydrogenation and defluorination occurred during degradation. Finally, the effects of the type and position of functional groups on the reaction rate and defluorination yield were investigated via multivariable linear regression analysis. Notably, the electron-withdrawing nature of functional groups appeared to have a greater impact on the defluorination yield of fluoroarenes than the calculated C-F bond dissociation energy.
Asunto(s)
Rodio , Catálisis , Rodio/química , Cinética , Halogenación , Oxidación-Reducción , Fluorobencenos/química , Hidrocarburos Fluorados/químicaRESUMEN
BACKGROUND: The SCHUMANN study evaluated the efficacy and safety of the selective P2 × 3 antagonist eliapixant in patients with endometriosis-associated pelvic pain (EAPP). METHODS: SCHUMANN was a randomized, placebo- and active comparator-controlled, double-blind to placebo and open-label to comparator, parallel-group, multicenter, dose-finding phase 2b study. The participants were women with surgically diagnosed endometriosis who fulfilled defined EAPP criteria. Participants were randomized 1:1:1:1 to twice daily (BID) 25 mg, 75 mg, or 150 mg oral eliapixant or a placebo for 12 weeks. An exploratory once-daily elagolix 150 mg treatment group was also included. The primary endpoint was the absolute change in mean worst EAPP from baseline to the end of intervention (EOI). RESULTS: Overall, 215 participants were randomized for treatment (44 to eliapixant 25 mg, 44 to eliapixant 75 mg, 43 to eliapixant 150 mg, 43 to a placebo, and 41 to elagolix 150 mg). For safety reasons, the study was terminated early; both treatment and enrollment stopped immediately, producing less than 50% of the planned number of completers. The study found no significant differences in EAPP reduction from baseline between groups and no significant dose-response model. The elagolix 150 mg group showed better pain reduction than any of the other groups. No new safety signals were observed, relative to the previously known safety profile of eliapixant, which was generally well tolerated. However, one case of moderate and probably drug-induced liver injury in a participant receiving eliapixant 150 mg BID supported the association between eliapixant and a potential increase in liver function values, defined before the start of the phase 2 program. CONCLUSIONS: This study did not meet its primary objective as no statistically significant or clinically relevant differences in changes of mean worst EAPP from baseline were observed between treatment groups. The single observed case of moderate, probably drug-induced liver injury was the second case in the eliapixant phase 2 program conducted in the following indications: refractory or unexplained chronic cough, diabetic neuropathic pain, overactive bladder, and EAPP. Due to this, the benefit-risk ratio for the study was no longer considered to be positive. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov identifier NCT04614246; registered November 3, 2020.
Asunto(s)
Endometriosis , Dolor Pélvico , Humanos , Femenino , Endometriosis/complicaciones , Endometriosis/tratamiento farmacológico , Dolor Pélvico/tratamiento farmacológico , Dolor Pélvico/etiología , Adulto , Método Doble Ciego , Resultado del Tratamiento , Persona de Mediana Edad , Hidrocarburos Fluorados/uso terapéutico , Hidrocarburos Fluorados/efectos adversos , Relación Dosis-Respuesta a Droga , Dimensión del Dolor , PirimidinasRESUMEN
Ion-pairing reversed-phase liquid chromatography was utilized for the analysis of native and phosphorothioated oligonucleotides differing in the length (2-6mers and 21mer) and the number and position of phosphorothioate modifications. We investigated the influence of counterion (acetate vs. hexafluoroisopropanol) on the adsorption of eleven alkylamines on the stationary phases. A stronger adsorption of charged alkylamines on octadecyl- and phenyl-based stationary phases led to greater retention of oligonucleotides, and the adsorption of alkylamines was promoted with greater concentration of hexafluoroisopropanol in the mobile phase. Selected amines (triethylamine, dipropylamine, hexylamine) were used to study the resolution of n and n-x mers (main peak and its impurities shortened at 5´end), and diastereomeric separation of phosphorothioated oligonucleotides. The results confirmed a crucial role of alkylamine and counterion choice on the diastereomeric separation. The increasing hydrophobicity of alkylamine led to diminished diastereomeric selectivity which produced narrower phosphorothioated oligonucleotides peaks and led to improved n/n-x separation. Using hexafluoroisopropanol instead of acetate as counterion further enhances this effect (except for 100 mM concentration of hexafluoroisopropanol in combination with highly hydrophobic hexylamine). The elevated column temperature led to suppression of the diastereomeric resolution and improved resolution of n and n-x mers oligonucleotides. Baseline separation of oligonucleotides with different number of phosphorothioate linkages was achieved; this may be useful for therapeutic oligonucleotide analysis.
Asunto(s)
Cromatografía de Fase Inversa , Oligonucleótidos Fosforotioatos , Cromatografía de Fase Inversa/métodos , Oligonucleótidos Fosforotioatos/química , Oligonucleótidos Fosforotioatos/aislamiento & purificación , Estereoisomerismo , Aminas/química , Interacciones Hidrofóbicas e Hidrofílicas , Propanoles/química , Adsorción , Hidrocarburos FluoradosRESUMEN
Fluorinated organic chemicals, such as per- and polyfluorinated alkyl substances (PFAS) and fluorinated pesticides, are both broadly useful and unusually long-lived. To combat problems related to the accumulation of these compounds, microbial PFAS and organofluorine degradation and biosynthesis of less-fluorinated replacement chemicals are under intense study. Both efforts are undermined by the substantial toxicity of fluoride, an anion that powerfully inhibits metabolism. Microorganisms have contended with environmental mineral fluoride over evolutionary time, evolving a suite of detoxification mechanisms. In this perspective, we synthesize emerging ideas on microbial defluorination/fluorination and fluoride resistance mechanisms and identify best approaches for bioengineering new approaches for degrading and making organofluorine compounds.
Asunto(s)
Bacterias , Biodegradación Ambiental , Bioingeniería , Fluoruros , Fluoruros/metabolismo , Bioingeniería/métodos , Bacterias/metabolismo , Bacterias/efectos de los fármacos , Bacterias/genética , Halogenación , Hidrocarburos Fluorados/metabolismo , Hidrocarburos Fluorados/farmacologíaRESUMEN
Formally described in 2009, Phytophthora sansomeana is a pathogen of increasing interest in native, agricultural, and horticulturally important plant species. The objective of this study was to elucidate the symptomatic and asymptomatic host range of P. sansomeana on six agricultural crop species commonly used in field crop rotations in Michigan. In addition, sensitivity to oomicides commonly used in seed treatments, including oxathiapiprolin, mefenoxam, ethaboxam, and pyraclostrobin, was performed to aid in disease management recommendations. Plant biomass, quantity of P. sansomeana DNA in roots, and reisolations were used to assess pathogenicity and virulence of 18 isolates of P. sansomeana on each plant species using an inoculated seedling growth chamber assay. Isolates displayed varying levels of virulence to the hosts tested. Reisolations were completed for each plant species tested, and varying quantities of P. sansomeana DNA were found within all plant species root samples. Corn, wheat, soybean, dry bean, and winter cereal rye plants were symptomatic hosts with significant reduction observed in the total plant biomass. No significant reduction in total plant biomass was observed in oats, and oat roots harbored the least amount of P. sansomeana DNA. No P. sansomeana isolates were insensitive to the oomicide compounds tested with mean absolute inhibition (EC50) values of fungicide required for 50% growth inhibition values of 7.8 × 10-2 µg/ml for mefenoxam, 1.13 × 10-1 µg/ml for ethaboxam, 2.6 × 10-2 µg/ml for oxathiapiprolin, and 3.04 × 10-1 µg/ml for pyraclostrobin. These results suggest that common crop rotations in Michigan may not be a viable option to reduce soilborne inoculum accumulation and oomicide seed treatments could be considered for early-season management of P. sansomeana.
Asunto(s)
Avena , Glycine max , Phytophthora , Enfermedades de las Plantas , Secale , Semillas , Triticum , Zea mays , Phytophthora/efectos de los fármacos , Phytophthora/fisiología , Phytophthora/genética , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/prevención & control , Secale/microbiología , Zea mays/microbiología , Avena/microbiología , Triticum/microbiología , Semillas/microbiología , Glycine max/microbiología , Especificidad del Huésped , Fungicidas Industriales/farmacología , Estrobilurinas/farmacología , Raíces de Plantas/microbiología , Virulencia , Productos Agrícolas/microbiología , Michigan , Plantones/microbiología , Biomasa , Carbamatos/farmacología , Piridinas , Benzamidas , Alanina/análogos & derivados , Hidrocarburos Fluorados , PirazolesRESUMEN
The physiological mechanism of bone tissue regeneration is intricately organized and involves several cell types, intracellular, and extracellular molecular signaling networks. To overcome the drawbacks of autografts and allografts, a number of synthetically produced scaffolds have been manufactured by integrating ceramics, polymers, and their hybrid-composites. Considering the fact that natural bone is composed primarily of collagen and hydroxyapatite, ceramic-polymer composite materials seem to be the most viable alternative to bone implants. Here, in this experimental study, copolymer PVDF-TrFE has been amalgamated with HA ceramics to produce composite scaffolds as bone implants. In order to fabricate PVDF-TrFE-HA (polyvinylidene fluoride-trifluoroethylene-hydroxyapatite) composite scaffolds, solvent casting-particulate leaching technique was devised. Two scaffold specimens were produced, with different PVDF-TrFE and HA molar ratios (70:30 and 50:50), and then electrically polarized to observe the subsequent polarization impact on the tissue growth and the suppression of bacterial cell proliferation. Both the specimens underwent characterization to analyze their biocompatibility and bactericidal activities. The bacterial culture of Pseudomonas aeruginosa (P. aeruginosa) and Staphylococcus aureus (S. aureus) bacteria on the composites was studied to understand the antibacterial characteristics. Moreover, MG63 cells cultured on these as-formed composites provided information about osteogenesis. Improved osteogenesis and antibacterial efficacy were observed on both the composites. However, the composite with 70 wt% PVDF-TrFE and 30 wt% HA showed a higher bactericidal effect as well as osteogenesis. It was found that PVDF-TrFE-HA-based biomaterials have the potential for bone tissue engineering applications.
Asunto(s)
Durapatita , Polivinilos , Andamios del Tejido , Durapatita/química , Durapatita/farmacología , Andamios del Tejido/química , Polivinilos/química , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/crecimiento & desarrollo , Ensayo de Materiales , Humanos , Hidrocarburos Fluorados/química , Hidrocarburos Fluorados/farmacología , Antibacterianos/farmacología , Polímeros de FluorocarbonoRESUMEN
All areas of the modern society are affected by fluorine chemistry. In particular, fluorine plays an important role in medical, pharmaceutical and agrochemical sciences. Amongst various fluoro-organic compounds, trifluoromethyl (CF3) group is valuable in applications such as pharmaceuticals, agrochemicals and industrial chemicals. In the present study, following the strict OECD modelling principles, a quantitative structure-toxicity relationship (QSTR) modelling for the rat acute oral toxicity of trifluoromethyl compounds (TFMs) was established by genetic algorithm-multiple linear regression (GA-MLR) approach. All developed models were evaluated by various state-of-the-art validation metrics and the OECD principles. The best QSTR model included nine easily interpretable 2D molecular descriptors with clear physical and chemical significance. The mechanistic interpretation showed that the atom-type electro-topological state indices, molecular connectivity, ionization potential, lipophilicity and some autocorrelation coefficients are the main factors contributing to the acute oral toxicity of TFMs against rats. To validate that the selected 2D descriptors can effectively characterize the toxicity, we performed the chemical read-across analysis. We also compared the best QSTR model with public OPERA tool to demonstrate the reliability of the predictions. To further improve the prediction range of the QSTR model, we performed the consensus modelling. Finally, the optimum QSTR model was utilized to predict a true external set containing many untested/unknown TFMs for the first time. Overall, the developed model contributes to a more comprehensive safety assessment approach for novel CF3-containing pharmaceuticals or chemicals, reducing unnecessary chemical synthesis whilst saving the development cost of new drugs.