Tsugio Kaneko

A conformational restriction approach to the development of dual inhibitors of acetylcholinesterase and serotonin transporter as potential agents for Alzheimer's disease.

Alzheimers disease (AD) has been treated with acetylcholinesterase (AChE) inhibitors such as donepezil. However, the clinical usefulness of AChE inhibitors is limited mainly due to their adverse peripheral effects. Depression seen in AD patients has been treated with serotonin transporter (SERT) inhibitors. We considered that combining SERT and AChE inhibition could improve the clinical usefulness of AChE inhibitors. In a previous paper, we found a potential dual inhibitor, 1, of AChE (IC50=101 nM) and SERT (IC50=42 nM), but its AChE inhibition activity was less than donepezil (IC50=10 nM). Here, we report the conformationally restricted (R)-18a considerably enhanced inhibitory activity against AChE (IC50=14 nM) and SERT (IC50=6 nM).

Design, synthesis and structure-activity relationships of dual inhibitors of acetylcholinesterase and serotonin transporter as potential agents for Alzheimer's disease.

We have designed and synthesized a dual inhibitor of acetylcholinesterase (AChE) and serotonin transporter (SERT) as a novel class of treatment drugs for Alzheimers disease on the basis of a hypothetical model of the AChE active site. Dual inhibitions of AChE and SERT would bring about greater therapeutic effects than AChE inhibition alone and avoid adverse peripheral effects caused by excessive AChE inhibition. Compound (S)-6j exhibited potent inhibitory activities against AChE (IC(50)=101 nM) and SERT (IC(50)=42 nM). Furthermore, (S)-6j showed inhibitory activities of both AChE and SERT in mice brain following oral administration.

Paired pulse facilitation of GABAergic IPSCs in ventral horn neurons in neonatal rat spinal cord

Whole-cell patch-clamp recording of GABAergic inhibitory postsynaptic currents (IPSCs) were made in ventral horn neurons of neonatal rat lumbar spinal cord in slice. In contrast to the hippocampus where paired pulse depression is reported to be observed for GABAergic IPSCs, double pulse stimulation of GABAergic inputs resulted in enhancement in the amplitude of the second IPSC in the spinal ventral horn. The facilitation ratio was decreased during enhanced synaptic transmission by increasing Ca2+ concentration in the external recording solution. Baclofen and adenosine. which are reported to depress synaptic transmission by presynaptic mechanisms, depressed IPSCs and increased the facilitation ratio. A postsynaptic manipulation such as application of bicuculline or changing the driving force did not affect the facilitation ratio. These results suggest that paired pulse facilitation of GABAergic IPSCs observed in neonatal rat spinal ventral horn appears to be based upon a mechanism similar to that underlying frequency-dependent facilitation of excitatory synaptic transmission, and is sensitive to presynaptic changes in synaptic strength.

Is picolinic acid a glycine agonist at strychnine-sensitive receptors?

By means of unit recording and electrophoretic application, the effect of picolinic acid on feline spinal interneurons in situ was studied in comparison with glycine. Picolinic acid inhibited neuronal firing in 60% of neurons and in some cases the inhibitory actions were antagonized by strychnine. Inhibition of firing by glycine, which was also strychnine-sensitive, was reduced in case of concomitant administration of picolinic acid. These results suggest that picolinic acid might act as a glycine agonist at strychnine-sensitive receptors.

Enhancement of recurrent inhibition of the spinal monosynaptic reflex by preceding stimulation of the medullary raphe´in rats

Recurrent inhibition of the spinal monosynaptic reflex (MSR) elicited by conditioning stimulation of the ventral root in anesthetized rats was weaker than both the recurrent inhibition of the disynaptic reflex and the inhibition of the MSR elicited by conditioning stimulation of the adjacent dorsal root. Among these 3 inhibitions, the recurrent inhibition of the MSR was enhanced to a markedly greater extent by a preceding stimulation of the medullary raphé nucleus than were the other inhibitions. The magnitude of the enhancement of the recurrent inhibition of MSR also was much greater when the medullary stimulation was delivered 20 ms prior to the ventral root activation, as compared with a 30-ms interval. Recurrent inhibition of the MSR was enhanced by intravenous injection of lysergic acid diethylamide (LSD); however, the enhanced effect on recurrent inhibition elicited by stimulation of the raphé nucleus was not attenuated by the drug. These results suggest that there is a non-serotonergic, descending pathway which is capable of modulating motor output solely by means of recurrent inhibition of the MSR.

Picolinic acid and indole-2-carboxylic acid: two types of glycinergic compounds modulate motor function differentially.

1. A putative agonist for the strychnine-sensitive glycine receptor picolinic acid was tested for its anticonvulsant activities in mice and muscle-relaxant activities in rats and compared with indole-2-carboxylic acid (I2CA), an antagonist for the strychnine-insensitive glycine receptor. Their effects on segmental reflexes in the cat spinal cord were examined to elucidate their sites of action. 2. Picolinic acid (200 and 400 mg/kg IP) delayed the onsets of strychnine- but not pentylenetetrazole-induced seizures. It delayed the onsets of bicuculline-induced seizures only at the higher dose. I2CA (200 and 400 mg/kg IP) delayed the onsets of these 3 kinds of seizures. Both compounds reduced muscle tone in rat decerebrate rigidity at a dose of 100 mg/kg IV. 3. Picolinate methylester, a picolinate derivative with higher lipophilicity, depressed spinal reflexes in both intact and spinalized cats at cumulative doses of 25 to 200 mg/kg IV. I2CA (50 mg/kg IV) inhibited spinal reflexes only in intact preparations. 4. These results suggest that the anticonvulsant and muscle-relaxant activities of picolinic acid (PA) are due to inhibition of spinal neurons, but that I2CA selectively affects supraspinal structures.

LSD but not methysergide reduces the inhibitory effect of the medullary raphe stimulation on the spinal reflex in rats

Conditioning stimulation of the nucleus raphe in the rat medulla enhanced the monosynaptic reflex (MSR) and depressed the polysynaptic reflex (PSR) in separate time courses. The PSR decreasing effect was reduced in preparations pretreated with reserpine, PCPA, or 5,6-DHT. The MSR increasing effect was not altered by these three pretreatments. Intravenously administered LSD but not methysergide reduced the PSR decreasing effect with no influence on the MSR increasing effect. These two drugs had similar effects on the unconditioned responses; the MSR was decreased and the PSR was increased. Thus, the PSR decreasing effect seems to be mediated via a serotonergic pathway probably descending from the nucleus raphe.

Central activation by CS-932, a functionally relative M1 agonist

NOBUYUKI KaSAWAl, KAZUKO WATANABE2, YOKO YAMAWAKI’, IKUKO NAGATSU ‘AND MINORU ONOZUKA3 lDept. of gnat., Fujita Health Univ. Sch. of Med., Toyoake. 470-l 101, Japan; Depts. of 2Physiology and 3Anat., Gifu Univ. Sch of Med., Gifi.r 500-8076, Japan To analyze age-associated changes in monoamine-biosynthesizing ability, we studied senescence-accelerated mice (SAM-P/& 8 months old) and accelerated senescence-resistant mice (SAM-R/l,8 months old) as controls. We administered, intraperitoneahy, 2, 4-diamino-&hydroxypyrimidine (DAHP) or parachlorophenylalanine (PCPA) to both groups of mice and measured the time course of changes in dopamine (DA)immunoreactivity (IR) in the nigrostriatal system, and serotonm (5-HT)IR in the raphe dorsalis in PAP-stained sections using a microphotometry system (Luzex FS, Nicon). While DA-IR in the nigrostriatal system of SAM-R/l was lowest at 3 h after the administration of DAHP and recovered to normal value 24 h after, that of the SAM-p/8 was lowest at 6 h following DAHP administration and did not recover even after 72 h. With respect to 5-HT-IR, while that in SAM-R/l following PCPA administration was lowest at 24 h after the administrauon and recovered 48-72 h after, that in SAM-P/8 showed almost no change at 24 h after and decreased slightly 48-72 h after the administrationThese studies strongly suggest that in SAM-P&while catecholaminergic neurons are significantly affected by monoamine synthesis inhibitors,serotonergic neurons do not show significant changes.

Preferential enhancement of inhibitory synaptic transmission by CS-722 in the ventral horn neurons of neonatal rat spinal cord

We studied the modulatory effect of (R)-4-chloro-2-(2-hydroxy-3-morpholinopropyl)-5-phenyl-4-isoxaz olin-3-one hydrochloride (CS-722), a centrally acting muscle relaxant, on synaptic transmission in the ventral horn neurons of neonatal rat lumbar spinal cord in slices using whole cell recording techniques. Pharmacologically isolated excitatory or inhibitory postsynaptic currents (EPSCs and IPSCs) were evoked by stimulating neighboring neurons. CS-722 preferentially enhanced IPSCs with little effects on EPSCs. Moreover, CS-722 reduced paired pulse facilitation of gamma-aminobutyric acid (GABA)-mediated IPSCs, suggesting its action on the presynaptic terminal. Preferential facilitatory effects on inhibitory synaptic transmission seem to be one of the mechanisms underlying muscle relaxation of this compound.