New Drugs 2020, part 2.

This article reviews seven drugs recently approved by the FDA, including indications, precautions, adverse reactions, and nursing considerations.

Quantification of 4-aminopyridine in plasma by capillary electrophoresis with electrokinetic injection.

A rapid and sensitive CE method for the determination of 4-aminopyridine in human plasma using 3,4-diaminopyridine as an internal standard was developed and validated. The analytes were extracted from 0.5-mL aliquots of human plasma by liquid-liquid extraction, using 8 mL of ethyl ether, and injected electrokinetically into capillary electrophoresis equipment. The instrumental conditions were obtained and optimized by Design of Experiments (DOE--factorial and response surface model), having as factors: separation voltage, ionic strength (buffer concentration), pH and temperature. The response variables were migration time, resolution, tailing factor and drug peak area. After obtaining mathematically predicted values for the response variables with best factors combinations, these were reproduced experimentally in good agreement with predicted values. In addition to optimal separation conditions obtained by Design of Experiments, sensitivity was improved using electrokinetic injection at 10 kV for 10 s, and a capillary with 50 cm effective length and 100 microm I.D. The final instrumental conditions were voltage at 19 kV, capillary temperature at 15 degrees C, wavelength at 254 nm, and phosphate buffer 100 mM, pH 2.5 as the background electrolyte. This assay was linear over a concentration range of 2.5-80 ng/mL with a lower limit of quantification of 2.5 ng/mL. The relative standard deviation for the assay precision was <7% and the accuracy was >95%. This method was successfully applied to the quantification of 4-aminopyridine (4-AP) in plasma samples from patients with spinal cord injury.

K(+)-channel blockers restore synaptic plasticity in the neuromuscular junction of dunce, a Drosophila learning and memory mutant.

The effects of K(+)-channel blockers on synaptic transmission in dunce (dnc), a Drosophila learning and memory mutant, were investigated. Larvae dnc mutants lack facilitation and post-tetanic potentiation (PTP) at their motor end-plates; dnc mutants are also deficient in a form of phosphodiesterase, and exhibit abnormally high levels of cyclic adenosine 3',5'-monophosphate (cAMP). A two-microelectrode voltage-clamp was used to record end-plate currents and spontaneous end-plate currents from longitudinal ventrolateral third-instar larval muscle. The K(+)-channel blockers 3,4-diaminopyridine (3,4-DAP) and tetraethylammonium (TEA), at micromolar concentrations, caused a reversible decrease in end-plate current amplitudes both in wild-type and mutant end-plates. In the presence of blockers, a period of high-frequency stimulation (tetanus) of the nerve gave way to a transient increase in the end-plate currents of dnc mutants resembling facilitation and PTP in normal end-plates; 3,4-DAP and TEA also restored facilitation and PTP in normal end-plates after incubation with a non-hydrolysable analogue of cAMP (8Br-cAMP). It is suggested that a specific K+ conductance might be relevant to the lack of synaptic plasticity at the dnc neuromuscular synapses.

Calcium action potentials in innervated and denervated rat muscle fibres.

We studied the generation of calcium action potentials (Ca APs) in innervated and denervated fibres of the extensor digitorum longus of the rat in a tetraethylammonium (TEA) sulphate solution plus 3-4 diaminopiridine (3-4 DAP). The main results are the following: (1) more than 90% of the innervated fibres were capable of developing well-sustained Ca APs that were blocked by Cd or nifedipine; (2) the incidence of Ca APs obtained from the denervated fibres was substantially lower than in the control preparations; (3) no relation was found between the appearance of Ca APs in the denervated fibres and the resting membrane potential (Vm), stimulus duration (500-2000 ms) or holding potential (-80, -100 mV); (4) The percentage of denervated fibres that exhibited Ca APs was increased significantly with the following procedures. First, by raising the external Ca concentration to 14 mM; second, by depleting the intracellular K concentration by overnight exposure of the muscles to a free K-Cs solution; (c) and third, by incubating the muscles in 500 nM apamin, a venom that inhibits the K conductance activated by Ca. Several factors may be involved in the lower incidence of Ca APs obtained in denervated fibres: (1) a diminished Ca current due to a reduction in the driving force as a result of an increment in the intracellular Ca concentration; (2) a persistence of a shunting K conductance that is not inhibited by TEA and 3-4 DAP; (3) a shift in the voltage dependence of the activation and inactivation parameters of the Ca current or the appearance of a new type of Ca channel with a different kinetics.

The effects of pentachlorophenol (PCP) at the toad neuromuscular junction.

1. Effects of PCP at the frog neuromuscular junction were studied in vitro in sciatic nerve sartorius muscle of the toad Pleurodema-thaul. 2. Within the concentration 0.003-0.1 mM, PCP caused a dose-time-dependent block of evoked transmitter release acompanied by an increase in the rate of spontaneous quantal release. 3. PCP induced an increase in miniature endplate potential (MEPP) frequency and it was not antagonized in a Ca2(+)-free medium, indicating that it does not depend upon Ca2+ influx from the external medium, but may act by releasing Ca2+ from intraterminal stores. 4. The present data, together with previous results concerning PCP at eighth sympathetic ganglia indicate that 3,4-diaminopyridine (3,4-DAP) counteracts the effects of PCP on synaptic transmission. This result suggests that PCP interfering Ca2+ influx occurs during depolarization of motor nerve terminals.

The actions of phenol and pentachlorophenol (PCP) on axonal conduction, ganglionic synaptic transmission, and the effect of pH changes.

1. A comparison of phenol, pentachlorophenol (PCP) and procaine effects on axonal conduction were studied in vitro in the sciatic nerves of toad. PCP and procaine were respectively 6.3 and 3.15 times more potent than phenol in blocking axonal conduction. 2. Effects of PCP on synaptic transmission were studied in vitro in the eighth sympathetic ganglion of toad. 3. Axonal conduction block and synaptic transmission block by phenol was reversible, but not that by PCP. 4. When the PCP ionization was increased, a lesser per cent reached the site of action, reducing its capacity to block the axonal conduction and ganglionic transmission. 5. PCP plus, 3,4-Diaminopyridine (3,4-DAP) decreased synaptic transmission block from post-ganglionic compound action potential (CAP) responses to supramaximal preganglionic stimulation.

Relation between action potential duration and mechanical activity on rat diaphragm fibers. Effects of 3,4-diaminopyridine and tetraethylammonium.

The aim of this work was to study the electrical and mechanical properties of small bundles of rat diaphragm muscle treated with two blockers of the delayed potassium rectification channels: 3,4-diaminopyridine (3,4-DAP, 2.5 mM) and tetraethylammonium (TEA, 20 mM). Twitch tension was significantly potentiated by TEA and 3,4-DAP (39% and 59% respectively). Maximal tetanic tension was not affected by both drugs. The voltage dependence of the tension vs the resting membrane potential was shifted to lower values in TEA and 3,4-DAP. 3,4-DAP increased the caffeine contracture tension (2.5-10 mM) and lowered the caffeine contracture threshold. The duration of the action potential (measured at the level of -40 mV) was increased by TEA and 3,4-DAP solutions. This change was a consequence of the decrease in the rate of repolarization of the action potential. In addition, TEA reduced the amplitude and the rate of rise of the action potential. We suggested that the increment in the duration of the action potential and the shift of the mechanical threshold to more negative values of membrane potential might be the factors involved in the twitch potentiation induced by the TEA and 3,4-DAP solutions.