Efficacy, safety, and dose-response characteristics of glipizide gastrointestinal therapeutic system on glycemic control and insulin secretion in NIDDM. Results of two multicenter, randomized, placebo-controlled clinical trials. The Glipizide Gastrointestinal Therapeutic System Study Group.

OBJECTIVE: To investigate the efficacy, safety, and dose-response characteristics of an extended-release preparation of glipizide using the gastrointestinal therapeutic system (GITS) on plasma glucose, glycosylated hemoglobin (HbA1c), and insulin secretion to a liquid-mixed meal in NIDDM patients. RESEARCH DESIGN AND METHODS: Two prospective, randomized, double-blind, placebo-controlled, multicenter clinical trials were performed in 22 sites and 347 patients with NIDDM (aged 59 +/- 0.6 years; BMI, 29 +/- 0.3 kg/m2; known diabetes duration, 8 +/- 0.4 years) were studied. Each clinical trial had a duration of 16 weeks with a 1-week washout, 3-week single-blind placebo phase, 4-week titration to a fixed dose, and 8-week maintenance phase at the assigned dose. In the first trial, once-daily doses of 5, 20, 40, or 60 mg glipizide GITS were compared with placebo in 143 patients. In the second trial, doses of 5, 10, 15, or 20 mg of glipizide GITS were compared with placebo in 204 patients. HbA1c, fasting plasma glucose (FPG), insulin, C-peptide, and glipizide levels were determined at regular intervals throughout the study. Postprandial plasma glucose (PPG), insulin, and C-peptide also were determined at 1 and 2 h after a mixed meal (Sustacal). RESULTS: All doses of glipizide GITS in both trials produced significant reductions from placebo in FPG (range -57 to -74 mg/dl) and HbA1c (range -1.50 to -1.82%). Pharmacodynamic analysis indicated a significant relationship between plasma glipizide concentration and reduction in FPG and HbA1c over a dose range of 5-60 mg, with maximal efficacy achieved at a dose of 20 mg for FPG and at 5 mg for HbA1c. PPG levels were significantly lower, and both postprandial insulin and C-peptide levels significantly higher in patients treated with glipizide GITS compared with placebo. The percent reduction in FPG was comparable across patients with diverse demographic and clinical characteristics, including those with entry FPG > or = 250 mg/dl, resulting in greater absolute decreases in FPG and HbA1c in patients with the most severe hyperglycemia. Despite the forced titration to a randomly assigned dose, only 11 patients in both studies discontinued therapy because of hypoglycemia. Glipizide GITS did not alter lipids levels or produce weight gain. CONCLUSIONS: The once-daily glipizide GITS 1) lowered HbA1c, FPG, and PPG over a dose range of 5-60 mg, 2) was maximally effective at 5 mg (using HbA1c) or 20 mg (using FPG) based on pharmacokinetic and pharmacodynamic relationships, 3) maintained its effectiveness in poorly controlled patients (those with entry FPG > or = 250 mg/dl), 4) was safe and well tolerated in a wide variety of patients with NIDDM, and 5) did not produce weight gain or adversely affect lipids.

Effect of chronic treatments with tandospirone and imipramine on serotonin-mediated behavioral responses and monoamine receptors.

The effects of acute and chronic treatment of rats with the tricyclic antidepressant imipramine, the 5-HT1A receptor partial agonist tandospirone, or its metabolite 1-PP were compared on behavioral responses produced by the activation of 5-HT receptors and on brain monoamine receptors. The behaviors examined were the 5-HT behavioral syndrome elicited by the 5-HT1A receptor agonist 8-OH-DPAT and the head shake response produced by the 5-HT2 receptor agonist DOB. Drug treatments were administered either by subcutaneous infusion from implanted minipumps or by repeated injection and the effects of chronic drug treatment were assessed when the drug was present and absent at the time of testing. The infusion of tandospirone blocked elicitation of the 5-HT behavioral syndrome when tested after 1 or 14 days of drug treatment (drug present) and 24 hr after the drug was withdrawn (drug absent). When administered by injection, tandospirone blocked the production of the 5-HT syndrome 1 hr (drug present), but not 24 hr (drug absent), following either 1 day or 14 days of drug treatment. Chronic infusion of imipramine did not alter the 5-HT syndrome. Chronic, but not acute, injections of imipramine blocked the 5-HT syndrome when tested 1 hr but not 24 hr, after the final injection. Treatment with 1-PP did not alter the 5-HT syndrome. The head shake response was attenuated by acute and chronic injection of tandospirone either 1 or 24 hr after treatment, although chronic infusion of tandospirone did not alter this behavior. Head shaking was attenuated by the infusion and injection of imipramine after acute treatment, chronic treatment, or following drug withdrawal. Chronic injection of 1-PP also inhibited the head shake response 24 hr after injection, although 1-PP was ineffective at all other times and when given by infusion. The density of hippocampal 5-HT1A receptors was unaltered by the chronic drug treatments. 5-HT2 receptor density in frontal cortex was reduced by the chronic infusion of either tandospirone, imipramine, and 1-PP, but only by chronic injections of imipramine. The density of cortical beta-adrenergic receptors was reduced following chronic imipramine injections or infusion. The results suggest that both tandospirone and imipramine may regulate 5-HT-mediated responses and 5-HT2 receptor density, which may contribute to their efficacy as antidepressants, although their effects were dependent upon the method of administration and may involve different neuropharmacological mechanisms.

Specific 1,25-dihydroxyvitamin D3 binding sites in choroid plexus.

Quantitative autoradiographic analysis of [3H] 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) binding in vitamin D deficient mice provided evidence for high levels of specific binding in choroid plexus and, to a lesser extent, ventral hippocampus. Sucrose gradient analysis yielded a 3-4S peak of specific [3H]1,25(OH)2D3 binding in bovine choroid plexus, but not amygdala or hippocampus. Scatchard analysis of [3H]1,25(OH)2D3 binding in bovine choroid plexus yielded KD = 0.23 +/- 0.06 nM and Nmax = 43.5 +/- 0 fmol/g tissue (n = 5). This result indicates the presence of significant receptor-like [3H]1,25(OH)2D3 binding sites in the choroid plexus and, thus, suggests roles for this hormone in regulating the entry of calcium into the brain and/or in the central regulation of calcium homeostasis.

Sertraline and cocaine-induced locomotion in mice. I. Acute studies.

The present study assessed the behavioral and pharmacokinetic interaction between the serotonin uptake blocker sertraline and cocaine in C57BL/6ByJ mice. Pretreatment with sertraline (1-32 mg/kg IP) did not affect the total amount of spontaneous locomotor activity during 50 min following administration of cocaine (15-40 mg/kg IP). At doses of sertraline (16 and 32 mg/kg) much higher than those found to inhibit ex vivo neuronal uptake of serotonin by 50% (1-2 mg/kg), the peak of cocaine-induced locomotor activity was shifted towards a later time. A similar effect was seen after pretreatment with serotonin uptake blockers other than sertraline, and also after desipramine. Sertraline (16 and 32 mg/kg), given 60 min prior to cocaine, did not affect levels of cocaine in brain and plasma, and cocaine administration did not alter the brain level of sertraline. Although female mice were more responsive to cocaine than male mice, they were not different in their response to sertraline.

Sertraline and cocaine-induced locomotion in mice. II. Chronic studies.

The effects of repeated treatment with the serotonin uptake blocker sertraline on cocaine-induced locomotion in female C57BL/6ByJ mice were examined in three paradigms. First, when animals were treated for 2 weeks with a daily injection of 8 mg/kg IP of sertraline (or placebo) and challenged with cocaine (25 mg/kg IP) 1 h after the final sertraline injection, their cocaine-induced locomotion was the same as that of placebo-pretreated controls. Second, animals were treated for 2 weeks with cocaine (25 mg/kg IP once a day) (or saline) and then for 2 weeks with sertraline (8 mg/kg IP once a day) (or placebo). Locomotion induced by cocaine (25 mg/kg IP) administered 1 h after the final sertraline (placebo) injection was higher in cocaine- than saline-pretreated mice (sensitization), but there was no difference between sertraline- and placebo-pretreated animals. Third, daily treatment with sertraline (8 mg/kg IP) did not change the locomotor stimulatory effect of cocaine (25 mg/kg IP) administered after a 3-week continuous infusion of cocaine (22 mg/kg/day SC) by osmotic minipumps or after three, four, or seven injections of cocaine (15 or 25 mg/kg IP). After cocaine administration (25 mg/kg IP), animals pretreated repeatedly with sertraline (8 mg/kg IP once a day for 2 weeks) had the same plasma or brain levels of cocaine as those pretreated with placebo; there was no difference between cocaine- and saline-treated mice in brain levels of sertraline or desmethylsertraline.

Effects of 5,7-dihydroxytryptamine on serotonin1 and serotonin2 receptors throughout the rat central nervous system using quantitative autoradiography.

The effects of the serotonin neurotoxin 5,7-dihydroxytryptamine (5,7-DHT), on serotonin1 (5-HT1) and 5-HT2 receptors were investigated using the high degree of resolution provided by quantitative autoradiography in an effort to determine the synaptic location of these receptors. 5,7-DHT treatment resulted in a decrease in 5-HT1 binding in the dentate gyrus and CA3c/4 of the anterior hippocampus and in the dorsal raphe nucleus, whereas no changes were observed in the posterior hippocampus nor in many other brain structures. 5-HT2 receptors exhibited no changes in any brain area examined in response to 5,7-DHT treatment, despite over 90% serotonin depletion in most of the forebrain nuclei examined. The results indicate that at least some of the 5-HT1 sites labelled by [3H]5-HT in the hippocampus and dorsal raphe nucleus are presynaptic, whereas 5-HT2 receptors are probably postsynaptic. In addition, the distribution profiles of 5-HT1 and 5-HT2 binding sites were compared in the rat central nervous system at various anatomical levels. 5-HT1 binding sites were identified using [3H]5-HT, while 5-HT2 binding sites were labelled with [3H]ketanserin. Both receptor subtypes displayed distinctly different localization patterns, which, in most cases was the inverse of the other pattern. In the brainstem it is significant that 5-HT2 receptors are concentrated in the facial nucleus and the motor nucleus of the trigeminal nerve, areas known to influence head and facial movement. The serotonin-mediated head-shake response occurs when 5-HT2 receptors are activated. In contrast, 5-HT1 receptors are distributed throughout the brainstem and in specific portions of the spinal cord. These areas are thought to control the serotonin behavioral syndrome and this behavior is 5-HT1A-mediated. All raphe nuclei were devoid of 5-HT2 receptors; only 5-HT1 receptor were found in these nuclei. Correlations with serotonin terminal distribution patterns are discussed. The pattern of 5-HT2 receptor distribution was also compared with the pattern of alpha 1 receptors, using [3H]prazosin in order to determine whether [3H]ketanserin significantly labels alpha 1 receptors. Although some similarities exist, overlap of binding did not occur in other nuclei, indicating that alpha 1 contamination of this system is probably negligible.

Adrenal steroid modulation of vasoactive intestinal peptide effect on serotonin1 binding sites in the rat brain shown by in vitro quantitative autoradiography.

In the present work we demonstrate by means of quantitative in vitro autoradiography that the vasoactive intestinal peptide (VIP) is able to increase the number of serotonin1 (5-HT1) binding sites in the dorsal subiculum of the rat hippocampus and to decrease them in the suprachiasmatic nucleus (SCN). Bilateral adrenalectomy (ADX) for 6 days counteracted the stimulatory effect of VIP on 5-HT1 binding sites in the dorsal subiculum, but did not modify the inhibitory effect of the peptide in the SCN. Moreover, ADX increased 5-HT1 binding sites in response to VIP in various subfields of the hippocampus as well as in the superior colliculus and in the dorsal lateral septum, but this effect was not observed in normal or in ADX rats bearing a corticosterone implant. The present data are suggestive of a possible interaction between VIP and 5-HT in the regulation of the SCN and of a modulatory role of adrenal steroids in VIP activity in the hippocampal formation.

Sex steroid modulation of the serotonin behavioral syndrome.

The sex difference observed in frequency of rats exhibiting the serotonin behavioral syndrome induced by pargyline/1-tryptophan depends on hormonal state. Castration eliminated the sex difference in drug response in adult and prepubertal males, whereas ovariectomy had little effect. Dihydrotestosterone administration to males (10-30 days) reinstated the sex difference, but had little effect in females. Testicular feminized mutants (Tfm/y), deficient in androgen receptors, respond like females. Estrogen administration has no effect in either sex. Manipulation of the hormonal environment on postnatal days 0-7 (blockade of aromatization in males or estradiol administration to females) has no effect on the expression of the sex difference when the animals were tested as adults. Therefore, androgens acting via androgen receptors appear to mediate this subsensitivity of male rats to the drug challenge. The results of these experiments indicate that sex and hormonal environment are important variables in determining the experimental and perhaps clinical responses to drugs.

Modulation by vasoactive intestinal peptide (VIP) of serotonin receptors in membranes from rat hippocampus.

The vasoactive intestinal peptide (VIP) has been located in various structures of the rat brain, but few actions of the peptide have been reported as yet. Because VIP might interact with classical neurotransmitter systems in the central nervous system as it does in the periphery, we investigated whether VIP can modulate serotonin (5-HT1) receptors in membrane preparations obtained from brain areas which contain various amounts of VIP and 5-HT receptors. The presence of bacitracin alone, which protects VIP from proteolytic degradation, decreases the affinity of [3H]5-HT binding in almost all of the structures tested. Scatchard analysis indicates that, in the presence of bacitracin, VIP significantly decreases the affinity and increases the number of specific high affinity binding sites for [3H]5-HT in the dorsal hippocampus. VIP induces a dose-dependent increase in the number of 5-HT1 receptors with a maximal response of 60% with 10(-7) M VIP. At the same concentration, neither secretin nor glucagon modifies 5-HT1 receptor density. No effect of VIP is observed in the ventral hippocampus, parietal cortex, whole hypothalamus, and midbrain. This effect of VIP is not observed when bacitracin is omitted, and the presence of calcium ions does not alter the efficacy of the VIP effect. No effect of VIP is obtained on [3H]spiperone binding assayed with 10 microM mianserin to define specific binding. The present data suggest that some of the effects of 5-HT in the hippocampus may be modulated by VIP.

Sex differences in serotonin 1 receptor binding in rat brain.

Male and female rats exhibit sex differences in binding by serotonin 1 receptors in discrete areas of the brain, some of which have been implicated in the control of ovulation and of gonadotropin release. The sex-specific changes in binding, which occur in response to the same hormonal (estrogenic) stimulus, are due to changes in the number of binding sites. Castration alone also affects the number of binding sites in certain areas. The results lead to the conclusion that peripheral hormones modulate binding by serotonin 1 receptors. The status of the serotonin receptor system may affect the reproductive capacity of an organism and may be related to sex-linked emotional disturbances in humans.

Modulation by vasoactive intestinal peptide of serotonin receptors in the dorsal hippocampus of the rat brain: an autoradiographic study.

Brain sections of 32 microns from the hippocampus were incubated with tritiated serotonin (5-HT) in the presence or absence of 10(-7) M vasoactive intestinal peptide (VIP). Sections were run for biochemical Scatchard analysis or quantitative autoradiography by means of LKB 3H-Ultrofilm. On sections from dorsal hippocampus, VIP increases the amount of 5-HT1 receptors and decreases the affinity for the ligand. Densities measurements show that this effect is located in a discrete area of the hippocampus, the dorsal subiculum. The present data suggest that some of the neurotransmitter effects of 5-HT in the central nervous system can be modulated by VIP.

Serotonin receptor modulation by estrogen in discrete brain nuclei.

The effect of estradiol on serotonin (5HT1) receptors of ovariectomized female rats was studied in discrete brain nuclei by a micro-binding assay and quantitative autoradiography. The two methods show similar distribution of the 5HT1 receptor and estradiol significantly increases the density of receptors in the preoptic area, anterior hypothalamus, lateral septum and arcuate-median eminence. The increase in 5HT1 receptors in these particular nuclei may be related to estrogen control of ovulation and/or other estrogen-mediated central phenomena.

Inhibition of lordosis behavior by intrahypothalamic implants of pargyline.

Hypothalamic sites responsible for monoaminergic of gonadal hormone-facilitated female sexual behavior (the lordosis response) were investigated. Pargyline, an irreversible inhibitor of monoamine oxidase, was applied stereotaxically to the hypothalamus of ovariectomized, estrogen-primed females 2 h prior to progesterone administration. Application of pargyline dorsal to or within the lateral aspect of the ventromedial nucleus led to a reduction in lordosis quotients and quality scores 5-7 and 29-31 h later. Implantation dorsal to or within the dorsomedial nucleus did not inhibit lordosis responding 5-7 h later but did inhibit the response 29-31 h later. In both implant sites, lordosis responding returned to prepargyline levels within 55 h after drug placement. The effects of the pargyline cannulae were verified biochemically by measuring activity of monoamine oxidase in preoptic-hypothalamic nuclei. Enzyme activity was inhibited to some extent in all nuclei sampled. The ability of the implants to antagonize lordosis responding was related to the extent to which they inhibited monoamine oxidase activity in the hypothalamus. Results suggest that localized perturbations in hypothalamic cells whose monoamine metabolism is known to be affected by gonadal hormones is sufficient to affect female sexual behavior.

Temporary desynchronization among circadian rhythms with lateral fornix ablation.

Lateral or medial fornix suction ablations were performed on adult male rats in order to selectively ablate or leave intact, respectively, fibers which terminate in the region of the suprachiasmatic nucleus and hypophysiotropic area of the hypothalamus. Plasma adrenal corticosteroid secretion, locomotor activity, body temperature, and food and water intake were recorded at 4 h intervals over a period of 48 h in individual animals 7-10 days postoperatively. Lateral fornix ablation specifically disrupted adrenal corticosteroid periodicity. A least-squares spectrum analysis of the data indicated that corticosteroid may be under ultradian control after this lesion. All animals, regardless of treatment, exhibited normal circadian locomotor activity patterns. Aberrations in feeding, drinking and body temperature rhythms were occasionally observed. This represents a temporary dissociation between the rhythmic expression of corticosteroid secretion and activity, temperature, feeding and drinking. The evidence presented lends support to the multi-oscillator theory of circadian organization, and suggests that the anteroventral subiculum, via the medial corticohypothalamic tract, is important in the regulation of some, but not all, circadian parameters. In addition to the observations on the rhythmicity of locomotor activity, the extent to which the animals are active is also significantly different between groups; ie., the hyperactivity of fornix-transected animals previously reported by others was found to be associated with lateral and not medial fornix ablation.

Differential fornix ablations and the circadian rhythmicity of adrenal corticosteroid secretion.

Suction ablations of the medial or lateral fornix were performed in order to transect selectively the medial corticohypothalamic tract (mcht) which originates in the anteroventral subiculum and travels in the lateral fornix terminating in the basal hypothalamus. The circadian rhythmicity of plasma adrenal corticosteroid levels was assessed in individual animals 1--2 weeks postoperatively. Ablation of the lateral fornix disrupted the periodicity of corticosteroid secretion which is normally synchronized with the light--dark cycle, whereas medial fornix ablation or neocortical ablation caused no such disruption. Group mean levels of plasma adrenal corticoids were higher in the lateral fornix-ablated animals than in the medial fornix-ablated, neocortically ablated, or intact control animals. These findings suggest that the anteroventral subiculum is important in the regulation of adrenal corticosteroid rhythmicity, and that it exerts an inhibitory influence upon corticosteroid release.