KM-408, a novel phenoxyalkyl derivative as a potential anticonvulsant and analgesic compound for the treatment of neuropathic pain.

BACKGROUND: Epilepsy frequently coexists with neuropathic pain. Our approach is based on the search for active compounds with multitarget profiles beneficial in terms of potential side effects and on the implementation of screening for potential multidirectional central activity. METHODS: Compounds were synthesized by means of chemical synthesis. After antiseizure and neurotoxicity screening in vivo, KM-408 and its enantiomers were chosen for analgesic activity evaluations. Further safety studies included acute toxicity in mice, the effect on normal electrocardiogram and on blood pressure in rats, whole body plethysmography in rats, and in vitro and biochemical assays. Pharmacokinetics has been studied in rats after iv and po administration. Metabolism has been studied in vivo in rat serum and urine. Radioligand binding studies were performed as part of the mechanism of action investigation. RESULTS: Selected results for KM-408: Ki sigma = 7.2*10-8; Ki 5-HT1A = 8.0*10-7; ED50 MES (mice, ip) = 13.3 mg/kg; formalin test (I phase, mice, ip)-active at 30 mg/kg; SNL (rats, ip)-active at 6 mg/kg; STZ-induced pain (mice, ip)-active at 1 mg/kg (von Frey) and 10 mg/kg (hot plate); hot plate test (mice, ip)-active at 30 mg/kg; ED50 capsaicin test (mice, ip) = 18.99 mg/kg; tail immersion test (mice)-active at 0.5%; corneal anesthesia (guinea pigs)-active at 0.125%; infiltration anesthesia (guinea pigs)-active at 0.125%. CONCLUSIONS: Within the presented study a novel compound, R,S-2-((2-(2-chloro-6-methylphenoxy)ethyl)amino)butan-1-ol hydrochloride (KM-408) with dual antiseizure and analgesic activity has been developed for potential use in neuropathic pain treatment.

Ligand assisted CuAAC labelling and RP-HPLC analysis of zidovudine and Retrovir using propargyl-Fmoc probe.

The extensive application of zidovudine (ZDV) as a stand-alone anti-HIV drug and a component in antiviral combination therapies, has made its analysis important both in the pharmaceutical and environmental context. The azide group in ZDV structure makes it a ready-to-use substrate for copper-catalyzed azide-alkyne cycloaddition (CuAAC), which is an efficient method for "click chemistry" labeling. In this paper, we describe a ligand-assisted CuAAC procedure for the precolumn derivatization of ZDV. We used propargyl-Fmoc fluorescent label and trans-2-(4-((dimethylamino)methyl)-1H-1,2,3-triazol-1-yl)cyclohexan-1-ol (AMTC) as a copper-binding ligand. We tested the applicability of AMTC for precolumn derivatization and developed chromatographic analytical procedures for ZDV and its formulation (50 mg/5 ml oral solution, Retrovir™ syrup). Our research aimed to improve labeling efficiency with a Cu-chelating ligand, using an accessible and affordable fluorescent probe. We also developed a sustainable mechanochemical synthesis procedure for obtaining propargyl-Fmoc in a gram scale and thus boosted the accessibility of this probe. The advantages of the developed derivatization procedure are its simplicity and easy availability of the propargyl-Fmoc probe. Moreover, the high lipophilicity of the propargyl-Fmoc probe enables efficient separation of the analyte from polar matrix components. In addition, the derivatization procedure can be performed directly on a sample solution. We tested its usability for samples in environmental and biological matrices, including tap water, river water, urine, and human serum.

Multigram-scale enzymatic kinetic resolution of trans-2-azidocyclohexyl acetate and chiral reversed-phase HPLC analysis of trans-2-azidocyclohexanol.

Lipase-catalyzed hydrolytic kinetic resolution is a method of obtaining optically pure chiral alcohols and amines, which requires additional tools for determining enantiomerical purity. Herein, we present a study on multigram-scale hydrolytic kinetic resolution of trans-2-azidocyclohexyl acetate using Pseudomonas cepacia lipase immobilized on Immobead support. We investigated several parameters of the preparative-scale process: temperature, organic co-solvent, and the influence of calcium ions. Moreover, we have developed an efficient fluorenylmethyloxycarbonyl chloride (Fmoc-Cl) derivatization protocol for 2-azidocyclohexanol, which enabled chiral reversed-phase high-performance liquid chromatography (RP-HPLC) determination of enantiomeric excess.