In Vitro Antileishmanial and Antischistosomal Activities of Anemonin Isolated from the Fresh Leaves of Ranunculus multifidus Forsk

Leishmaniasis and schistosomiasis are neglected tropical diseases (NTDs) infecting the world’s poorest populations. Effectiveness of the current antileishmanial and antischistosomal therapies are significantly declining, which calls for an urgent need of new effective and safe drugs. In Ethiopia fresh leaves of Ranunculus multifidus Forsk. are traditionally used for the treatment of various ailments including leishmaniasis and eradication of intestinal worms. In the current study, anemonin isolated from the fresh leaves of R. multifidus was assessed for its in vitro antileishmanial and antischistosomal activities. Anemonin was isolated from the hydro-distilled extract of the leaves of R. multifidus. Antileishmanial activity was assessed on clinical isolates of the promastigote and amastigote forms of Leishmania aethiopica and L. donovani clinical isolates. Resazurin reduction assay was used to determine antipromastigote activity, while macrophages were employed for antiamastigote and cytotoxicity assays. Antischistosomal assays were performed against adult Schistosoma mansoni and newly transformed schistosomules (NTS). Anemonin displayed significant antileishmanial activity with IC₅₀ values of 1.33 nM and 1.58 nM against promastigotes and 1.24 nM and 1.91 nM against amastigotes of L. aethiopica and L. donovani, respectively. It also showed moderate activity against adult S. mansoni and NTS (49% activity against adult S. mansoni at 10 µM and 41% activity against NTS at 1 µM). The results obtained in this investigation indicate that anemonin has the potential to be used as a template for designing novel antileishmanial and antischistosomal pharmacophores.     

Development and Validation of a Simple,Selective, and Accurate Reversed-Phase Liquid Chromatographic Method with Diode Array Detection (RP-HPLC/DAD) for the Simultaneous Analysis of 18 Free Amino Acids in Topical Formulations

Even though there are reported methods for the quantification of free amino acids (FAAs) in biological products, no work has been done on the analysis of these substances in formulations. Moreover, further research is required as the reported methods do not fulfill analytical method requirements. The objective of this study was, therefore, to develop and validate a rapid, reliable, and appropriate RP-HPLC/DAD method for the simultaneous determination of 18 FAAs (l-Ala, l-Arg, l-Asn, l-Asp, l-Gln, l-Glu, l-Gly, l-His, l-Ile, l-Lue, l-Lys, l-Met, l-Orn, l-Phe, l-Pro, l-Ser, l-Thr, and l-Val) in topical formulations. After appropriate method development, the technique was validated for selectivity, linearity and range, limit of detection, limit of quantification, precision, and accuracy. The samples were derivatized with 9-fluorenylmethyl chloroformate (Fmoc-Cl). Chromatographic separation was performed on InfinityLab Poroshell 120 E.C 18 (3?×?50) mm, 2.7 μm column at 25 °C. The mobile phase consisting of water and acetonitrile adjusted to appropriate pH was pumped in gradient mode at a flow rate of 0.7 mL/min. Ten microliters were injected and analyte detection was conducted using a DAD. The results indicate that the method was selective for these FAAs. It was linear over the concentration range of 5–80 µM with a correlation coefficient greater than 0.995. Moreover, it was sensitive, precise, accurate, and robust. All the reported drawbacks of RP-HPLC-based analysis of FAAs were resolved, and hence, this new method can be considered appropriate for the analysis of these FAAs in topical formulations.

     

Protonation sites and hydrogen bonding in mono-hydrobromide salts of two N,4-diheteroaryl 2-aminothiazoles

Structural Chemistry - The synthesis and structural characterization of N-(6-methoxypyridin-3-yl)-4-(pyridin-2-yl)thiazol-2-amine mono-hydrobromide monohydrate (3) and...     

Crystallographic evidence for unintended benzisothiazolinone 1‐oxide formation from benzothiazinones through oxidation

1,3‐Benzothiazin‐4‐ones (BTZs) are a promising new class of drugs with activity against Mycobacterium tuberculosis, which have already reached clinical trials. A product obtained in low yield upon treatment of 8‐nitro‐2‐(piperidin‐1‐yl)‐6‐(trifluoromethyl)‐4H‐benzothiazin‐4‐one with 3‐chloroperbenzoic acid, in analogy to a literature report describing the formation of sulfoxide and sulfone derived from BTZ043 [Tiwari et al. (2015). ACS Med. Chem. Lett. 6 , 128–133], is a ring‐contracted benzisothiazolinone (BIT) 1‐oxide, namely, 7‐nitro‐2‐(piperidine‐1‐carbonyl)‐5‐(trifluoromethyl)benzo[d]isothiazol‐3(2H)‐one 1‐oxide, C₁₄H₁₂F₃N₃O₅S, as revealed by X‐ray crystallography. Single‐crystal X‐ray analysis of the oxidation product originally assigned as BTZ043 sulfone provides clear evidence that the structure of the purported BTZ043 sulfone is likewise the corresponding BIT 1‐oxide, namely, 2‐[(S)‐2‐methyl‐1,4‐dioxa‐8‐azaspiro[4.5]decane‐8‐carbonyl]‐7‐nitro‐5‐(trifluoromethyl)benzo[d]isothiazol‐3(2H)‐one 1‐oxide, C₁₇H₁₆F₃N₃O₇S. A possible mechanism for the ring contraction affording the BIT 1‐oxides instead of the anticipated constitutionally isomeric BTZ sulfones and antimycobacterial activities thereof are discussed.     

Galvinoxyl radicals: Synthesis of new derivatives,determination of low oxygen contents,and stability studies

Two new derivatives of galvinoxyl (1), a perdeutered (2) and an adamantyl-analog (3) for potential applications as spin probes were synthesized. The synthesis with deuterated educts yielded 2 with 98% D. It exhibited an 18-line EPR spectrum in octanol with narrow peak-to-peak linewidth. The EPR spectrum of 3 was very similar to galvinoxyl, but with differences in the linewidth due to unresolved long-range couplings with adamantyl-protons. Compound 2 showed a higher response to oxygen (4.8 μT/% O₂) than 1 (2.8 μT/% O₂). The coupling pattern of 2 allowed the determination of oxygen at low levels (0–6%) by a new type of analysis of the EPR pattern. The stability of the radicals strongly depended on the amount of hydrogalvinoxyl, a by-product of the galvinoxyl synthesis, and the solvent type. A molecular mechanism for the stabilization by hydrogalvinoxyl and the influence of solvent type is proposed.     

Structural Characterization of Two Polymorphs of 1-(4-Methylpyridin-2-yl)thiourea and Two Derived 2-Aminothiazoles

Journal of Chemical Crystallography - Two polymorphic forms of 1-(4-methylpyridin-2-yl)thiourea (1) and the crystal and molecular structures of the 2-aminothiazoles...