Molecular Docking and GC/MS-Based Approach for Identification of Anxiolytic Alkaloids from Griffinia (Amaryllidaceae) Species in a Zebrafish Model.

Griffinia gardneriana Ravenna, Griffinia liboniana Morren and Griffinia nocturna Ravenna (Amarillydaceae) are bulbous plants found in tropical regions of Brazil. Our work aimed to determine the alkaloid profiles of Griffinia spp. and evaluate their anxiolytic potential through in vivo and in silico assays. The plants grown in greenhouses were dried and their ground bulbs were subjected to liquid-liquid partitions, resulting in alkaloid fractions that were analyzed by gas chromatography coupled to mass spectrometry (GC-MS). Anxiolytic activity was evaluated in zebrafish (Danio rerio) through intraperitoneal injection at doses of 40, 100 and 200 mg/kg in light-dark box test. GC-MS analyses revealed 23 alkaloids belonging to different skeleton types: lycorine, homolychorine, galanthamine, crinine, haemanthamine, montanine and narcisclasine. The chemical profiles were relatively similar, presenting 8 alkaloids common to the three species. The major component for G. gardneriana and G. liboniana was lycorine, while G. nocturna consisted mainly of anhydrolycorine. All three alkaloid fractions demonstrated anxiolytic effect. Furthermore, pre-treatment with diazepam and pizotifen drugs was able to reverse the anxiolytic action, indicating involving the GABAergic and serotonergic receptors. Molecular docking showed that the compounds vittatine, lycorine and 11,12-dehydro-2-methoxyassoanine had high affinity with both receptors, suggesting them to be responsible for the anxiolytic effect.

Synthesis, structural, characterization, antibacterial and antibiotic modifying activity, ADMET study, molecular docking and dynamics of chalcone (E)-1-(4-aminophenyl)-3-(4-nitrophenyl)prop-2-en-1-one in strains of Staphylococcus aureus carrying NorA and MepA efflux pumps.

Chalcones have an open chain flavonoid structure that can be obtained from natural sources or by synthesis and are widely distributed in fruits, vegetables, and tea. They have a simple and easy to handle structure due to the α-ß-unsaturated bridge responsible for most biological activities. The facility to synthesize chalcones combined with its efficient in combating serious bacterial infections make these compounds important agents in the fight against microorganisms. In this work, the chalcone (E)-1-(4-aminophenyl)-3-(4-nitrophenyl)prop-2-en-1-one (HDZPNB) was characterized by spectroscopy and electronic methods. In addition, microbiological tests were performed to investigate the modulator potential and efflux pump inhibition on S. aureus multi-resistant strains. The modulating effect of HDZPNB chalcone in association with the antibiotic norfloxacin, on the resistance of the S. aureus 1199 strain, resulted in increase the MIC. In addition, when HDZPNB was associated with ethidium bromide (EB), it caused an increase in the MIC value, thus not inhibiting the efflux pump. For the strain of S. aureus 1199B, carrying the NorA pump, the HDZPNB associated with norfloxacin showed no modulatory, and when the chalcone was used in association with EB, it had no inhibitory effect on the efflux pump. For the tested strain of S. aureus K2068, which carries the MepA pump, it can be observed that the chalcone together the antibiotic resulted in an increase the MIC. On the other hand, when chalcone was used in association with EB, it caused a decrease in bromide MIC, equal to the reduction caused by standard inhibitors. Thus, these results indicate that the HDZPNB could also act as an inhibitor of the S. aureus gene overexpressing pump MepA. The molecular docking reveals that chalcone has a good binding energies -7.9 for HDZPNB/MepA complexes, molecular dynamics simulations showed that Chalcone/MetA complexes showed good stability of the structure in an aqueous solution, and ADMET study showed that the chalcone has a good oral bioavailability, high passive permeability, low risk of efflux, low clearance rate and low toxic risk by ingestion. The microbiological tests show that the chalcone can be used as a possible inhibitor of the Mep A efflux pump.Communicated by Ramaswamy H. Sarma.

In vitro antiviral effect of ethanolic extracts from Azadirachta indica and Melia azedarach against goat lentivirus in colostrum and milk.

This study aimed to evaluate, in vitro, the use of leaf extracts of Azadirachta indica (A. indica) and Melia azedarach (M. azedarach) as antivirals against caprine lentivirus (CLV) in colostrum and milk of goat nannies. These were collected from eight individuals and infected with the standard strain of CLV. Samples were then subdivided into aliquots and treated with 150 µg/mL of crude extract, and with ethyl acetate and methanol fractions for 30, 60, and 90 min. Next, somatic cells from colostrum and milk were co-cultured with cells from the ovine third eyelid. After this step, viral titers of the supernatants collected from treatments with greater efficacy in co-culture were assessed. The organic ethyl acetate fractions of both plants at 90 min possibly inhibited the viral activity of CLV by up to a thousandfold in colostrum. In milk, this inhibition was up to 800 times for the respective Meliaceae. In conclusion, the ethanolic fraction of ethyl acetate from both plants demonstrated efficacy against CLV in samples from colostrum and milk when subjected to treatment, which was more effective in colostrum.

Synthesis and in silico study of chenodeoxycholic acid and its analogues as an alternative inhibitor of spike glycoprotein of SARS-CoV-2.

COVID-19, caused by SARS-CoV-2, is a viral infection that has generated one of the most significant health problems in the world. Spike glycoprotein is a crucial enzyme in viral replication and transcription mediation. There are reports in the literature on using bile acid in the fight against this virus through in vitro tests. This work presents the synthesis of nine chenodeoxycholic acid derivatives (1-9), which were prepared by oxidation, acetylation, formylation, and esterification reactions, and the analogs 6-9 have not yet been reported in the literature and the possibility of conducting an in silico study of bile acid derivatives as a therapeutic alternative to combat the virus using glycoprotein as a macromolecular target. As a result, five compounds (1, 6-9) possessed favorable competitive interactions with the lowest energies compared to the native ligand (BLA), and the highlighted compound 9 got the best scores. At the same time, analog 1 presented the best ADME filter result. Molecular dynamics also simulated these compounds to verify their stability within the active protein site to seek new therapeutic propositions to fight against the pandemic. Physical and spectroscopic data have fully characterized all the compounds.Communicated by Ramaswamy H. Sarma.

In vitro ovicidal effect of a Senecio brasiliensis extract and its fractions on Haemonchus contortus.

BACKGROUND: Haemonchosis affects sheep husbandry and its treatment is often compromised due to the development of anthelminthic resistance. Plant-derived bioactive compounds have been studied as alternative to control Haemonchus contortus. The objective of this study was to evaluate the effect of Senecio brasiliensis extracts on H. contortus egg hatching and infective larvae migration. RESULTS: The aqueous extract from dried and fresh plant and alkaloid-enriched fraction of the previously dried leaves of S. brasiliensis inhibited H. contortus egg hatching. The main plant compound in alkaloid fraction was integerrimine, a pyrrolizidine alkaloid (PA). However, the aqueous extract from dried plant displayed higher efficacy when compared to their alkaloid enriched or non-polar fractions, meaning that, although PAs contributed to the ovicidal effect, other compounds in the plant can also contribute to their effect. Furthermore, the aqueous extract from dried plant also had higher efficacy than aqueous extract from fresh plant in larvae migration inhibition. Finally, extract from dried plant presented low in vitro cytotoxic effect. CONCLUSION: Taken together our results suggest a good anthelmintic effect of S. brasiliensis, especially when aqueous extract is prepared from dried plant. Further in vivo studies should be performed focused on forms of administration of this extract in rearing sheep.

Development of a UPLC-ESI-MS method for simultaneous determination of flavonoids and diterpenes in Egletes viscosa (L.) Less herbal products.

Egletes viscosa (L.) Less (Asteraceae) is a tropical aromatic herb whose flower buds are used for gastrointestinal disorders. Its gastroprotective properties have been attributed to the flavonoid ternatin and the furan diterpenes tanabalin and centipedic acid. The aim of this study was to develop a method of ultra-performance liquid chromatography coupled to electrospray ionization and mass spectrometry (UPLC-ESI-MS) for identifying the constituents from E. viscosa flower buds as well as quantifying its bioactive compounds in herbal products. Infusions and tincture from wild and commercial E. viscosa materials were directly injected on a UPLC-quadrupole-time-of-flight system (UPLC-q-TOF). Afterwards, reference standards were used to quantify ternatin, tanabalin and centipedic acid in these samples, employing a UPLC-single quadrupole system set up for positive mode and Selected Ion Monitoring (SIM). Seventeen compounds were identified, including caffeic acid derivatives, flavonoids and diterpenes, from which seven have been reported for the first time in this specie. The quantification method showed good linearity (R > 0.99), accuracy, precision and sensitivity. The intra-day and inter-day precisions presented relative standard deviations inferior to 6.04 and 8.78%, respectively. The recoveries of all the analytes varied 82.25-117.87%. The limits of detection and quantification ranged between 10-25 µg/L and 25-75 µg/L, respectively. For the samples, the contents of ternatin, tanabalin and centipedic acid ranged from 0.89 to 8.03 mg/L, 0.84-16.8 mg/L and 3.21-16.8 mg/L, respectively. The method demonstrated to be rapid, sensitive and reliable for the quality control of E. viscosa-based products, besides being applicable to other plant extracts containing flavonoids and diterpenes.