William J. Jackson

Functional characterization of the novel neuronal nicotinic acetylcholine receptor ligand GTS-21 in vitro and in vivo

(2.4)-Dimethoxybenzylidene anabaseine dihydrochloride (GTS-21), a compound that interacts with rat neuronal nicotinic acetylcholine receptors (nAChRs), was evaluated using human recombinant nAChRs in vitro and various pharmacokinetic and behavioral models in rodents, dogs and monkeys. GTS-21 bound to human alpha 4 beta 2 nAChR (K1-20 nM) 100-fold more potently than to human alpha 7 nAChR, and was 18- and 2-fold less potent than (-)-nicotine at human alpha 4 beta 2 and alpha 7 nAChR, respectively. Functionally. GTS-21 stimulated [5H]dopamine release from rat striatal slices with an EC50 of 10 +/- 2 microM (250-fold less potent and 70% as efficacious as (-)-nicotine), an effect blocked by the nAChR antagonist dihydro-beta-erythroidine. However, GTS-21 did not stimulate human alpha 4 beta 2 nor human ganglionic nAChRs significantly. In vivo, GTS-21 had no adverse effect on dog blood pressure (< or = 2.5 micromol/kg i.v. bolus infusion), in marked contrast with (-)-nicotine, GTS-21 (-62 micromol/kg.s.e.) also did not cross-discriminate significantly with (-)-nicotine in rats and did not reduce temperature or locomotion in mice. Neither was it active in the elevated plus maze anxiety model (0.19-6.2 micromol/kg.IP) in normal mice. However, GTS-21 did improve learning performance of monkeys in the delayed matching-to-sample task (32-130 nmol/kg.i.m.).

Beneficial effects of nicotine administered prior to a delayed matching-to-sample task in young and aged monkeys

Our earlier studies have demonstrated that administration of low micrograms/kg doses of nicotine to young adult monkeys prior to a delayed matching-to-sample (DMTS) task resulted in a centrally mediated improvement in performance of the task, particularly when delay intervals which most greatly challenged the animals capabilities were involved. The present study confirmed these findings using a completely computer driven and automated procedure. In addition, performance on the DMTS was observed to be enhanced when animals were again tested 24 h after the dose of nicotine. Further analysis of the data indicated that the majority of enhancement to nicotine could be accounted for by a greatly increased performance at the least preferred stimulus color. Position preference (left vs. right stimulus) was not a factor in nicotine-induced enhancement. Two aged monkeys (34 years old Macaca mulatta) were significantly more difficult to train in the DMTS task and their longest delay capabilities were significantly shorter than the young animals (Macaca fascicularis). Nevertheless, the aged animals were essentially similar in most respects in their responses to nicotine administration. These data are consistent with a role for central nicotinic systems in memory performance and with the ability of nicotine to produce enhancement of selective features of mnemonic strategy in young and old monkeys. Furthermore, it is possible that either model, the aged animal, or the young animal stressed to his mnemonic capability may provide a good model for learning and memory disorders in humans.

Nicotine enhances delayed matching-to-sample performance by primates

The non-human primate provides an excellent model for studies of learning and memory, and one particular test, the delayed matching-to-sample task, is performed in a similar manner by both humans and non-human primates. Five young adult macaques were employed in this study, displaying variable capacities for retention in the task. Baseline performance was very consistent and three levels of performance difficulties (95-100%, 80-85% and 65-75% correct choices) were employed by including several delay intervals (0-60 sec) in each session. A reproducible enhancement in performance by nicotine in macaques performing a delayed matching-to-sample task was demonstrated. Nicotine enhanced performance with an average increase of 10% at the longest retention delay interval. This beneficial effect of nicotine was abolished in animals pretreated with a low dose (0.5 mg/kg) of mecamylamine to block central nicotinic receptors. Selective blockade of peripheral nicotinic receptors with hexamethonium was without effect on the nicotine response. A high dose (2 mg/kg) of mecamylamine itself induced a marked inhibition of performance, while an equivalent dose of hexamethonium was without effect. These experiments point to the possibility that central nicotinic receptors may be exploited pharmacologically to enhance memory performance. In this respect it is interesting that nicotine was most effective at enhancing performance when recall was more difficult, that is, on the longer retention interval delays. This could signify that nicotine might be particularly effective in the most impaired individuals. Lastly, it is encouraging that the mecamylamine induced decrease in cognitive performance might provide a new model of memory impairment from which to study the pathogenesis and develop new pharmacological strategies for the dementias.

Enhanced delayed matching performance in younger and older macaques administered the 5-HT4 receptor agonist, RS 17017.

Abstract Recent evidence indicates that the 5-HT4 subtype of serotonin receptor may modulate central cholinergic activity in regions of the mammalian CNS important to memory such as the frontal cortex, hippocampus and amygdala. These receptors could represent targets for drugs designed for the symptomatic therapy of Alzheimer’s disease (AD) and other disorders of memory. In the present study, the binding activity of RS 17017 (previously described as a selective 5-HT4 agonist) was assessed across a number of neurotransmitter receptors and binding sites, pharmacokinetic data were obtained, and the compound was evaluated in macaques for mnemonic effects via a computer-assisted delayed matching-to-sample task (DMTS). Binding data confirmed the 5-HT4 selectivity of the compound, while pharmacokinetic results revealed low oral bioavailability, but a large volume of distribution of the compound. Significant and reproducible improvements in DMTS accuracy were observed after oral administration of the compound across a dose-effect series in both younger and older monkeys. The results suggest that RS 17017 offers a potential for memory enhancement in disorders involving cognitive decline, and are consistent with a role for central 5-HT4 receptors in memory. Improvements in DMTS performance in aged monkeys may have particular implications for neurodegenerative conditions such as AD, whereas positive results in the younger monkeys indicate that RS 17017 (or similar compounds) may have additional potential in the therapeutics of memory disorders not necessarily associated with advanced age.

Clonidine enhances delayed matching-to-sample performance by young and aged monkeys

Clonidine, an alpha-2 noradrenergic agonist, has been shown to alter cognitive performance in humans and animals. Included among the evidence are studies which differ in their conclusions regarding the question of whether clonidine administration improves delayed response (DR) performance by nonhuman primates. The present results indicated that clonidine administration to both young and aged monkeys results in a modest performance improvement as measured by one of the commonly employed versions of DR performance-delayed matching-to-sample (DMTS). The clonidine-induced enhancement of DMTS had a duration of at least 24 h in both age groups.

Scopolamine reversal of nicotine enhanced delayed matching-to-sample performance in monkeys

The basis for the memory enhancing action of nicotine was evaluated in five adult monkeys (Macaca fascicularis) well trained in the performance of a delayed matching-to-sample (DMTS) paradigm. Nicotine (1.25-20 micrograms/kg, IM) produced a dose-dependent improvement in performance of the task. The optimal dose of nicotine for each monkey also improved performance when the animals were tested 24 h later in the no-drug situation. In the same animals, low doses of scopolamine produced a dose-dependent decrement in DMTS performance. A subthreshold dose (defined by DMTS performance decrement) of scopolamine was administered 20 min prior to the optimal dose of nicotine. Scopolamine pretreatment completely blocked the enhanced performance observed earlier with nicotine. The results of this study are consistent with the hypothesis that the enhanced cognitive performance associated with nicotine is due to central acetylcholine release and subsequent muscarinic receptor stimulation.

Improvement in accuracy of delayed recall in aged and non-aged, mature monkeys after intramuscular or transdermal administration of the CNS nicotinic receptor agonist ABT-418

Abstract ABT-418 was evaluated for its ability to enhance accuracy on a delayed matching-to-sample (DMTS) task by aged monkeys following intramuscular administration, and in non-aged mature monkeys following transdermal application. Aged monkeys were impaired in their performance of the DMTS task such that the longest delay intervals performed at above-chance levels extended only to 20 s. In contrast, for non-aged, mature animals, delay intervals extended to 140 s. In aged monkeys, the response to ABT-418 was highly individualized with animals responding to one or more doses in the range of 2–259 nmol/kg. A systematic dose-dependent enhancement of DMTS accuracy was not observed. When the individualized ”best dose” was administered on a separate occasion, overall DMTS accuracy was increased by 12.6%. By 24 h after administration, accuracy was at control levels. In young monkeys, a significant dose-dependent enhancement of DMTS performance (an overall increase of 11.25% above baseline accuracy) was observed 5 h after application of a transdermal patch designed to maintain steady-state plasma levels of ABT-418 of 40–60 ng/ml over a 24-h period. Again there was some individual responsiveness to one of the three doses. When data included only the individualized best doses of ABT-418 for each animal, a similar enhancement of accuracy was observed for both the 5-h and 24-h test intervals. In neither the aged nor the young cohorts was enhancement of performance associated with altered response latencies or with any overt side effects of ABT-418. Thus, these data are consistent with the ability of ABT-418 to improve DMTS performance in both young and aged monkeys. In aged monkeys, this response was observed only after administration of individualized optimal doses for different monkeys. In young monkeys, a more systematic enhancement of DMTS accuracy was observed. Further, transdermal delivery of ABT-418 in non-aged monkeys demonstrated prolonged performance enhancement compared with IM injection to at least 24 h after patch administration.

Delayed Matching-to-Sample in Monkeys as a Model for Learning and Memory Deficits: Role of Brain Nicotinic Receptors

The nervous system and behavioral repertoire of old world monkeys resembles the human neuro-behavioral system more than any other laboratory animal, except higher apes. In addition, spontaneous and conditioned behavior exhibited by the monkey is more similar to that of the human than any other laboratory animal1. Therefore, behavioral tasks which tap the higher cognitive abilities of these nonhuman primates may provide information more relevant to normal human aging and to the demential. The method most frequently employed to test the sophisticated cognitive repertoire of these monkeys has been one or another variation of the delayed response task. The delayed matching-to-sample (DMTS) task allows the measurement of abilities which are relevant to human aging, such as attention, strategy formation, reaction time in complex situations and memory for recent events. Thus, comparisons to human behavioral situations should involve less speculation than when lower animal subjects are employed. Interestingly, a similar version of this task has been employed to demonstrate cognitive impairment in Alzheimer’s Disease patients 2. The advent of the personal computer age has facilitated the automation of problem presentation and data collection associated with this task, and it is now practical to analyze DMTS performance at a more detailed level.

Cognitive effects of nicotinic cholinergic receptor agonists in nonhuman primates

The centrally acting cholinesterase inhibitor tacrine was compared with three nicotinic acetylcholine receptor (nAChR) agonists for their abilities to enhance performance of mature adult macaques performing a computer‐automated version of the delayed matching‐to‐sample (DMTS) task. All four drugs enhanced DMTS performance at one or more doses, although ABT‐418 [(S)‐3‐methyl‐5‐(1‐methyl‐2‐pyrrolidinyl) isoxazole] may be the most potent and the most effective of the four. Nicotine was less potent and less effective than ABT‐418 but was more potent than either tacrine or isoarecolone. At each animals respective maximally effective dose, task improvement ranged from approximately 14 to 30% over vehicle performance levels. Despite the significantly enhanced levels of performance improvement obtained on the day of drug administration, when the animals were tested 24 h later (in the absence of drug), only nicotine‐treated animals exhibited a significant improvement in performance. In an attempt to help explain this protracted improvement in DMTS performance to nicotine, cell surface nerve growth factor (NGF) receptors were measured in cultured PC‐12 cells before and after exposure to nicotine. Exposure to nicotine for 24 h resulted in a significant increase in cell surface NGF in the cells. However, even after nicotine was removed from the culture medium, NGF receptor protein continued to increase for an additional 24 h. The results of this study are consistent with the possibility that stimulation of central nAChRs may be employed to improve cognitive function in cognitively impaired individuals. They also suggest that one potential mechanism for the protracted beneficial effect of nicotine may involve the enhanced expression of brain NGF receptors. Drug Dev. Res. 38:196–203

Effects of stimulation or blockade of central nicotinic-cholinergic receptors on performance of a novel version of the rat stimulus discrimination task

This study evaluated the effects of two central nicotinic-cholinergic receptor agonists and an antagonist on performance accuracy of a rat, delayed stimulus discrimination task (DSDT). Rats were trained to discriminate between an auditory and visual stimulus by pressing a right or left lever. To diminish the rats ability to use mediating spatial strategies to solve the task, computer automated, retractable doors separated the animal from the levers during delay intervals, thus reducing positioning at the lever. After stable baselines were achieved, rats were grouped and administered placebo (saline) and nicotine, lobeline or mecamylamine in a randomized dose series. Each group received two complete series of the selected compound on different occasions. Mecamylamine impaired DSDT accuracy in a dose-dependent manner while optimal doses of nicotine and lobeline significantly improved accuracy. Nicotine differed from lobeline in regard to its interaction with a dose of mecamylamine (1.0 mg/kg) that had not impaired DSDT accuracy. Combined administration of lobeline and mecamylamine was followed by a significantly increased level of DSDT accuracy that was similar to the improvement following administration of lobeline alone. In contrast, combined administration of nicotine and mecamylamine did not result in increased DSDT accuracy. Furthermore, lobeline administration similarly improved accuracy of trials associated with both the light and the tone, while nicotine improved accuracy of trials associated with the light to a much greater degree. These data suggest that the increases in DSDT accuracy associated with lobeline may be expressed through non-nicotinic mechanisms or a nicotinic receptor which is not blocked by mecamylamine.