A novel and sensitive derivatization procedure for the determination of 2-cynaoacetamide in pharmaceutical samples using liquid chromatography with the fluorescence detection was discovered. The method is based on derivatization of 2-cynaoacetamide using 2-hydroxyacetophenone as a new derivatization reagent. The product of derivatization reaction was isolated and characterized using spectroscopic techniques namely LC-MS, NMR and IR. The structure of 2-cyanoacetamide derivative was unambiguously assigned as a 2-amino-4-phenylfuran-3-carboxamide. Two derivatization systems were optimized in terms of reaction temperature, reaction time, pH and concentration of 2-hydroxyacetophenone, and a new pre- and post-derivatization HPLC methods were developed. The separations on HPLC with pre-column derivatization were accomplished using stationary phase based on a XBridge C18 column (100×4.6, 3.5μm) and isocratic elution using the mobile phase acetonitrile - 0.1% formic acid (30:70, v/v). The separations on the HPLC with post-column derivatization were performed on stationary phase on a TSKgel Amide-80 column (150×4.6mm, 3μm). The mobile phase was a mixture of acetonitrile, methanol and 10mM sodium formate buffer at pH=4.5 in ratio 3:2:95 (v/v). Both HPLC methods were fully validated in terms of linearity, sensitivity (limit of detection and limit of quantification), accuracy and precision according to ICH guidelines. The pre-column derivatization method was linear in the range 1.1-2000μg/l with method accuracy≥98.2% and method precision RSD≤4.8%. The post-column derivatization method was linear in the range 12-2000μg/l. Method accuracy≥96.3% and method precision RSD≤3.5%. Proposed new methods were proved to be highly sensitive, simple and rapid, and were successfully applied to the determinations of 2-cynaoacetamide in pregabalin.

• Klíčová slova
• 2-cyanoacetamide, 2-hydroxyacetophenone, Derivatization, Fluorescence detection, Pregabalin,
• MeSH
• acetofenony chemie   MeSH
• hmotnostní spektrometrie MeSH
• indikátory a reagencie * MeSH
• magnetická rezonanční spektroskopie MeSH
• nitrily analýza   MeSH
• pregabalin chemie   MeSH
• spektroskopie infračervená s Fourierovou transformací MeSH
• vysokoúčinná kapalinová chromatografie metody   MeSH
• Publikační typ
• časopisecké články MeSH
• validační studie MeSH
• Názvy látek
• 2-cyanoacetamide MeSH Prohlížeč
• 2'-hydroxyacetophenone MeSH Prohlížeč
• acetofenony MeSH
• indikátory a reagencie * MeSH
• nitrily MeSH
• pregabalin MeSH

Viscosity-sensitive fluorophores, fluorescent molecular rotors based on aminobenzylidene-cyanoacetamide moiety, were tethered to 2'-deoxycytidine triphosphate via a propargylamine linker and incorporated into DNA by polymerases in primer extension, nicking enzyme amplification or PCR. DNA probes incorporating modified nucleosides show a light-up response upon binding to a protein.

• MeSH
• aniliny chemie   MeSH
• benzylidenové deriváty chemie   MeSH
• DNA vazebné proteiny chemie   MeSH
• DNA-dependentní DNA-polymerasy metabolismus   MeSH
• DNA chemie   MeSH
• fluorescenční barviva chemie   MeSH
• molekulární struktura MeSH
• nitrily chemie   MeSH
• vazba proteinů MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 2-cyanoacetamide MeSH Prohlížeč
• aniliny MeSH
• benzylidenové deriváty MeSH
• DNA vazebné proteiny MeSH
• DNA-dependentní DNA-polymerasy MeSH
• DNA MeSH
• fluorescenční barviva MeSH
• nitrily MeSH

OBJECTIVES: Both pyridine and pyrano derivatives have been previously shown to possess biologically relevant activity. In this study, we report the incorporation of these two scaffolds into one molecule. METHODS: The designed 3,3-dimethyl-6-oxopyrano[3,4-c]pyridines were synthesized by the acylation of enamine under Stork conditions followed by condensation of formed β-diketones with 2-cyanoacetamide. The structures of these compounds were confirmed by using a wide spectrum of physico-chemical methods. Their antiplatelet, anticoagulant and vasodilatory activity together with toxicity were evaluated. KEY FINDINGS: A series of 6-oxopyrano[3,4-c]pyridines 3a-j was obtained. Four of these compounds were reported for the first time. None of the tested compounds demonstrated anticoagulant effect but 8-methyl derivative (3a) was a potent antiplatelet compound with IC50 numerically twice as low as the clinically used acetylsalicylic acid. A series of further mechanistic tests showed that 3a interferes with calcium signaling. The compound is also not toxic and in addition possesses vasodilatory activity as well. CONCLUSIONS: Compound 3a is a promising inhibitor of platelet aggregation, whose mechanism of action should be studied in detail.

• Klíčová slova
• anticoagulant, antiplatelet activity, cytotoxicity, pyranopyridine, vasodilatory, β-diketones,
• MeSH
• agregace trombocytů * MeSH
• antikoagulancia farmakologie   MeSH
• inhibitory agregace trombocytů * chemie   farmakologie   MeSH
• pyridiny farmakologie   MeSH
• vztahy mezi strukturou a aktivitou MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antikoagulancia MeSH
• inhibitory agregace trombocytů * MeSH
• pyridiny MeSH