Isolation and Structural Elucidation of Impurity in Sulfonamide Intermediate of Glyburide

A reproducible isolation method by Reverse Phase (RP) preparative HPLC technique for the isolation of one crucial impurity at 1.65 RRT (Relative Retention Time) in sulfonamide stage of Glyburide API (Active Pharmaceuticals Ingredient) was developed. Preparative chromatography was done on Luna C8, 10µm (250 mm x 21.2mm) preparative HPLC column with acetonitrile: water in 70:30 % v/v proportion as a mobile phase and 8 ml/min as a flow rate. This impurity was detected at 300 nm UV-wavelength maximum. This impurity was isolated from synthesized crude impurity of sulfonamide stage of Glyburide substance by preparative HPLC by injecting 50 mg/ml concentration over 5 ml fixed loop. Isolated impurity was elucidated as N-methyl impurity of sulfonamide intermediate of Glyburide API by means of chromatographic and spectral data. Structural elucidation carried out by spectral data was reviewed. This impurity was analyzed by reverse phase HPLC for purity analysis. A Inertsil C8 (250 x 4.6) mm, 5µ particle size was employed for separation. The mobile phase consisted of Water: Acetonitrile: Methanol in the ratio of 60:15:25 % v/v. The flow rate was set at 1 ml/min. Detection was carried out at 300 nm. 10µL of 2 mg/ml concentration of sample in methanol was injected. The column oven temperature was at 25°C.

Studying on cytotoxic activity of ethyl acetate extracts and isolated substances from cultured Isaria cicadae F0004 in Vietnam against the MCF-7 cell lines and Jurkat cell lines

Isaria cicadae species of the Isaria genus have been isolated in many Asian countries, including China, Korea, Thailand, and Vietnam. Furthermore, Isaria cicadae content potential medicinal source in the prevention and treatment of cancer, strengthening the immune system. This study aims to investigate the cytotoxicity against MCF-7 and Jurkat T cell lines of the extract and isolated potential compounds from Isaria cicadae in Vietnam. The results showed that ethyl acetate (EA) extract from the fruit body of the Isaria cicadae F0004 strain had high cytotoxic activity against both MCF-7 and Jurkat cell lines at the concentration of 100 μg/mL with the IC50 value was reached 17.15 ± 1.68 and 10.37 ± 0.61 μg/mL respectively. The constituents of the EA extract from the fungus Isaria cicadae F0004 were isolated by column chromatography and preparative chromatography. Then, the structures were determined by spectroscopy 1H-NMR, 13C-NMR. It was obtained 5 compounds including uracil, 1-0-ethyl-ß-D-ribofuranose, ergosterol, p-hydroxybenzoic acid, protocatechuic acid. Protocatechuic acid isolated from Isaria cicadae F0004 that showed cytotoxic activity on MCF-7 and Jurkat cell lines with IC50 values of 5.97 ± 0.36 and 3.15 ± 0.64 (mM) respectively. This study is the basis for further research on the cytotoxic activity of the fungus Isaria cicadae F0004 in vitro.

Biological Activities of Extracts from Ageratum fastigiatum: Phytochemical Study and In Silico Target Fishing Approach

In the present study, the ethanolic extract from aerial parts of Ageratum fastigiatum was evaluated in vitro against epimastigote forms of Trypanosoma cruzi (Y strain), promastigote forms of Leishmania amazonensis (PH8 strain), and L. chagasi (BH400 strain). The extract was also evaluated against Staphylococcus aureus (ATCC 25 923), Escherichia coli (ATCC 11 775), Pseudomonas aeruginosa (ATCC 10 145), and Candida albicans (ATCC 36 802). The phytochemical screening was performed by thin-layer chromatography and high-performance liquid chromatography. The extract was fractionated using flash preparative chromatography. The ethanolic extract showed activity against T. cruzi, L. chagasi, and L. amazonensis and antimicrobial activity against S. aureus, E. coli, P. aeruginosa, and C. albicans. The phytochemical screening revealed coumarins, terpenes/sterols, and flavonoids in the ethanolic extract. In addition, the coumarin identified as ayapin was isolated from this extract. We also performed in silico prediction of potential biological activities and targets for compounds previously found in A. fastigiatum. Several predictions were confirmed both retrospectively and prospectively by experimental results described here or elsewhere. Some activities described in the in silico target fishing approach were validated by the ethnopharmacological use and known biological properties. Some new activities and/or targets were predicted and could guide future studies. These results suggest that A. fastigiatum can be an interesting source of substances with antiparasitic and antimicrobial activities.

Antileishmanial Potential of Berberine Alkaloids From Berberis glaucocarpa Roots: Molecular Docking Suggests Relevant Leishmania Protein Targets

Leishmaniases are a spectrum of poverty-linked neglected parasitic diseases that are endemic in 88 countries around the globe and affect millions of people every year. Currently available chemotherapeutic options are inadequate due to side effects, high cost, prolonged treatment, and parasite resistance. Thus, there is an existing need to develop new potent and safer leishmanicidal drugs. Considering the folkloric antiulcer and leishmanicidal use of the genus Berberis and its alkaloids, 5 reported alkaloids, namely berberine (1), palmatine (2), columbamine (3), 8-trichloromethyldihydroberberine (4), and jatrorrhizine (5), were isolated from the roots of Berberis glaucocarpa using classical (column and preparative chromatography) and modern isolation techniques (Sephadex LH-20). Their structures were elucidated and established from 1D and 2D spectroscopic data. The isolated alkaloids displayed excellent antileishmanial potential with IC50 values ranging from 1.50 to 2.56 µM: 1 (1.50 ± 0.53 µM), 2 (2.31 ± 0.37 µM), 3 (2.56 ± 0.48 µM), 4 (1.40 ± 0.90 µM), 5 (2.44 ± 1.34 µM). While the IC50 value for the standard drug (Amphotericin-B) was found to be 1.08 ± 0.95 µM. All of the isolated alkaloids displayed excellent antileishmanial potential as well as minimal cytotoxicity against THP-1 monocytic cells. Molecular docking analysis has revealed Leishmania N-myristoyl transferase, methionyl-tRNA synthetase, pteridine reductase 1, oligopeptidase B, tyrosyl-tRNA synthetase, and/or glycerol-3-phosphate dehydrogenase to be potential protein targets for the alkaloids.

Potency of bisresorcinol from Heliciopsis terminalis on skin aging: in vitro bioactivities and molecular interactions

Background A bisresorcinol was isolated as the main constituent of Heliciopsis terminalis’s trunk (Proteaceae). Recently, resorcinol is applied as an active whitening agent in various cosmetic products. Because of the structural mimic to resorcinol, benefits of the bisresorcinol as an aging-enzyme antagonist were demonstrated in this study. Methods The bisresorcinol was purified from the crude ethanolic extract of H. terminalis’s trunk by solvent extraction and preparative chromatography, respectively. Inhibitory activity on collagenase, elastase, and tyrosinase of the compound was investigated by using a different spectroscopic technique. Molecular docking was carried out to predict possible interactions of the substance around the enzyme active sites. Results The IC50 values on collagenase of the bisresorcinol and caffeic acid were 156.7 ± 0.7 and 308.9 ± 1.6 µmole L−1, respectively. For elastase activity, the IC50 of 33.2 ± 0.5 and 34.3 ± 0.3 µmole L−1 was respectively determined for the bisresorcinol and ursolic acid. The bisresorcinol was inhibitory to tyrosinase by exhibiting the IC50 of 22.8 µmole L−1, and that of 78.4 µmole L−1 was present for β-arbutin. The bisresorcinol bound to collagenase, elastase, and tyrosinase with the respective binding energies of −5.89, −5.69, and −6.57 kcal mol−1. These binding energies were in the same ranges of tested inhibitors. The aromatic phenol groups in the structure were responsible for principle as well as supporting binding interactions with enzymes. Hydrogen binding due to hydroxyl groups and π-related attractive forces from an aromatic ring(s) provided binding versatility to bisresorcinol. Conclusion The bisresorcinol purified from H. terminalis might be useful for inclusion in cosmetic products as an aging-enzyme antagonist.

Effect of Ethanolic Caesalpinia sappan Fraction on In Vitro Antiviral Activity against Porcine Reproductive and Respiratory Syndrome Virus

Porcine reproductive and respiratory syndrome virus (PRRSV) is a major epidemic in pig production, leading to economic losses in the pig industry worldwide. The use of medicinal plants with antiviral properties might be useful help to prevent and control PRRSV outbreaks. Caesalpinia sappan (CS) heartwood is an important herbal ingredient used in Thai folk medicine, possessing various biological activities, including antiviral activity. The present study focuses on the in vitro antiviral activity against PRRSV of a semi-purified fraction of ethanolic CS crude extract using preparative high-performance liquid chromatography. Qualification of the fractions illustrating positive antiviral activity was carried out with liquid chromatography–quadrupole time-of-flight mass spectrometry. The preparative chromatography separated the crude extract into six consecutive fractions, among which the first fraction showed potential antiviral activity by inhibiting PRRSV replication in a MARC-145 monolayer (virus titer 2.75 median tissue culture infective dose (TCID50)/mL (log10) vs. 9.50 median log10 TCID50/mL of the control) at 72 h post-infection, and this fraction included byakangelicin, brazilin, naringenin, and brazilein. These results provide useful information for further study to effectively develop the CS bioactive antiviral compounds against PRRSV as a feed additive or veterinary drug in the pig industry.

From the culture broth to the erythritol crystals: an opportunity for circular economy

The reduction of sugar intake by adults has been stated by the World Health Organization as an important strategy to reduce the risk of non-communicable diseases. Erythritol is a four-carbon sugar alcohol that is considered as a highly suitable substitution for sucrose. This review article covers approaches for the separate stages of the biotechnological production of erythritol from cultivation to the downstream section. The first part focuses on the cultivation stage and compares the yields of erythritol and arising by-products achieved with different types of substrates (commercial versus alternative ones). The reported numbers obtained with the most prominently used microorganisms in different cultivation methods (batch, fed-batch or continuous) are presented. The second part focuses on the downstream section and covers the applied technologies for cell removal, recovery, purification and concentration of erythritol crystals, namely centrifugation, membrane separation, ion and preparative chromatography, crystallization and drying. The final composition of the culture broth and the preparative chromatography separation performance were identified as critical points in the production of a high-purity erythritol fraction with a minimum amount of losses. During the review, the challenges for a biotechnological production of erythritol in a circular economy context are discussed, in particular regarding the usage of sustainable resources and minimizing waste streams. Key points • Substitution of sucrose by erythritol can be a step towards a healthier society • Biotechnological production of erythritol should follow a circular economy concept • Culture broth composition and preparative chromatography are keys for downstreaming • Substrate, mother liquor and nutrients are challenges for circular economy