tetragastrin and butyloxycarbonyl-tryptophyl-methionyl-aspartyl-phenylalaninamide

The influence of intraventricular cholecystokinin-8S (CCK-8S) and systemic N-t-Boc-Trp-Met-Asp-Phe-amide (Boc CCK-4) was evaluated in the acoustic and fear-potentiated startle paradigms in CD-1 mice. In the light + tone startle condition. CCK-8S increased startle 168 hr after administration, compared with saline. In the tone startle condition, CCK-8S decreased startle immediately and 24 hr after administration, compared with saline. Among nonshocked mice, CCK-8S increased startle at 48 and 168 hr, compared with saline. In the light + tone condition, 5 microg Boc-CCK-4 did not influence startle, whereas 15 microg Boc CCK-4 decreased startle immediately, 24 hr, and 48 hr following administration. Results demonstrate that antecedent environmental experiences interact with subsequent pharmacological challenges in provoking the temporal expression of alterations in startle magnitude.

Topics: Acoustic Stimulation; Animals; Fear; Infusions, Intravenous; Injections, Intraventricular; Male; Mice; Nootropic Agents; Reflex, Startle; Sincalide; Tetragastrin

This study investigated a role of cholecystokinin (CCK) in the anxiolytic-like action of morphine, an agonist of mu-opioid receptors, in the rat plus-maze model of anxiety. The acute administration of morphine (1 mg/kg) induced a significant increase of exploratory activity in the plus-maze, but did not affect the locomotor activity in the motility test. The higher dose of morphine (2.5 mg/kg) tended to decrease the locomotor activity and, therefore, did not cause the anxiolytic-like action in the plus-maze. The other drugs (naloxone, BOC-CCK-4, L-365,260) and their combinations with morphine (0.5-1 mg/kg) did not affect the locomotor activity of rats. The opioid antagonist naloxone itself (0.5 mg/kg) did not change the exploratory activity in the plus-maze, but potently antagonized the anxiolytic-like action of morphine (1 mg/kg). An agonist of CCK(B)receptors BOC-CCK-4 (1-50 microgram/kg) induced a dose-dependent anxiogenic-like action in the plus-maze. Nevertheless, only one dose of BOC-CCK-4 (10 microgram/kg) completely reversed the action of morphine. Also, one dose of CCK(B)receptor antagonist L-365,260 (10 microgram/kg) was effective to modify the behaviour of rats in the elevated plus-maze. Namely, this dose of L-365,260 increased the ratio between open and total arm entries, a behavioural measure believed to reflect the anxiolytic-like action in the elevated plus-maze. The combination of L-365,260 (100 microgram/kg) with the sub-effective dose of morphine (0.5 mg/kg) caused the anxiolytic-like action in the plus-maze not seen if the drugs were given alone. In conclusion, morphine induces a potent anxiolytic-like action in the elevated plus-maze and CCK is acting as an endogenous antagonist of this effect of morphine.

Topics: Analgesics, Opioid; Animals; Anti-Anxiety Agents; Anxiety; Benzodiazepinones; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Interactions; Fear; Injections, Subcutaneous; Locomotion; Male; Maze Learning; Morphine; Naloxone; Narcotic Antagonists; Phenylurea Compounds; Rats; Rats, Wistar; Receptor, Cholecystokinin B; Receptors, Cholecystokinin; Tetragastrin

This series of experiments examined the effects of the cholecystokinin (CCK) fragments Boc-CCK-4 and CCK-8s on memory, reinforcement and anxiety following unilateral injection into the central nucleus of the amygdala (CeA). In experiment 1, rats with chronically implanted cannulae were injected with CCK-8s or Boc-CCK-4 and were tested on a one-trial uphill avoidance task. Post-trial injection of 20 ng Boc-CCK-4 or 1 ng CCK-8s was found to improve the retention performance, whereas lower and higher doses had no effect. The hypermnestic effects of Boc-CCK-4 and CCK-8s were no longer evident when injection was performed 5 h, rather than immediately, after the learning trial. In experiment 2, the elevated plus-maze was used to gauge anxiogenous properties of intra-amygdala injections of Boc-CCK-4 and CCK-8s in memory-enhancing doses. The treatment with 20 ng Boc-CCK-4 and 1 ng CCK-8s did not influence the number of entries into and time spent on the open and enclosed arms of the maze as well as other anxiety-related behaviors. In experiment 3, possible reinforcing effects of the CCK-fragments were examined. After intra-amygdala injection of Boc-CCK-4 or CCK-8s in memory-enhancing doses the rats were placed into one of four restricted quadrants of a circular open field (closed corral) for a single conditioning trial. Subsequent tests for conditioned corral preference revealed no evidence for reinforcing or aversive effects of the CCK-fragments. In sum, these findings indicate that Boc-CCK-4 and CCK-8s facilitate memory processing upon injection into the CeA without exerting reinforcing or anxiogenous effects.

Topics: Amygdala; Animals; Anxiety; Avoidance Learning; Behavior, Animal; Conditioning, Psychological; Male; Maze Learning; Memory; Rats; Rats, Wistar; Reinforcement, Psychology; Sincalide; Tetragastrin

This study investigated the interplay of cholecystokinin (CCK) and endogenous opioid peptides in the regulation of anxiety. The acute administration of non-selective CCK agonist caerulein (1 and 5 microg/kg) and a selective CCK(B) receptor agonist BOC-CCK-4 (1, 10 and 50 microg/kg) induced a dose-dependent anxiogenic-like action in the plus-maze model of anxiety. BOC-CCK-4 displayed a similar efficacy with caerulein, indicating that the described effect was mediated via CCK(B) receptor subtype. The opioid antagonist naloxone itself (0.5 mg/kg) did not change the exploratory activity of rats in the plus-maze. However, the combination of naloxone with the sub-effective doses of caerulein (1 microg/kg) and BOC-CCK-4 (1 microg/kg) induced a significant inhibition of exploratory behaviour in rats. Accordingly, CCK and endogenous opioid peptides have an antagonistic role in the exploratory model of anxiety in rats.

Topics: Animals; Anxiety; Ceruletide; Cholecystokinin; Disease Models, Animal; Drug Synergism; Exploratory Behavior; Male; Naloxone; Narcotic Antagonists; Rats; Rats, Wistar; Receptor, Cholecystokinin B; Receptors, Cholecystokinin; Tetragastrin

We previously reported novel Boc-CCK-4 (Boc-Trp-Met-Asp-Phe-NH2) derivatives possessing the general structure Boc-Trp-Lys[N epsilon-CO-NH-(R-Ph)]-Asp-Phe-NH2 (Shiosaki et al. J. Med. Chem. 1991, 34, 2837-2842). In contrast to Boc-CCK-4, which is 70-fold selective for the CCK-B receptor, the modified lysine-bearing tetrapeptides were highly potent and selective full agonists at the CCK-A receptor. Further investigation of the structure-activity profile following modification of the substituted phenylurea moiety appended off the lysine revealed that moving certain substituents, e.g. nitro or acetyl, from the 2- or 3-position on the phenyl ring to the 4-position, a relatively minor and subtle structural modification within the tetrapeptide, resulted in loss of CCK-A receptor selectivity and development of a trend toward CCK-B selectivity. These tetrapeptides, e.g. Boc-Trp-Lys[N epsilon-CO-NH-(4-NO2-Ph)]-Asp-Phe-NH2 and Boc-Trp-Lys[N epsilon-CO-NH-(4-Ac-Ph)]-Asp-Phe-NH2, were full agonists relative to CCK-8 in stimulating intracellular calcium mobilization in a cell line that expresses the CCK-B receptor.

Topics: Cell Line; Magnetic Resonance Spectroscopy; Mass Spectrometry; Receptor, Cholecystokinin A; Receptor, Cholecystokinin B; Receptors, Cholecystokinin; Structure-Activity Relationship; Tetragastrin

Memory-modulating and reinforcing effects of the cholecystokinin (CCK) fragments, CCK-8S and Boc-CCK-4, after systemic application in rats were investigated. Habituation to the novelty of environmental stimuli was used to test for mnemonic effects using two different tasks (rearing behavior in an open field; head-dips in a hole-board). Immediate posttrial administration of CCK-8S and Boc-CCK-4 resulted in a reduction of rearing and head-dip behavior during testing, indicative of enhanced habituation and, thus, facilitation of memory. In contrast, administration of CCK-8S and Boc-CCK-4 with a delay of 2.5 or 5 h after training or pretrial injection of CCK-8S did not enhance habituation. No evidence for reinforcing or aversive properties of CCK-8S and Boc-CCK-4 was observed in a conditioned place preference task. In summary, the results indicate memory-enhancing effects of peripherally, posttrial-administered CCK-8S and Boc-CCK-4.

Topics: Amino Acid Sequence; Animals; Choice Behavior; Conditioning, Operant; Exploratory Behavior; Habituation, Psychophysiologic; Learning; Male; Memory; Molecular Sequence Data; Nootropic Agents; Rats; Rats, Wistar; Sincalide; Structure-Activity Relationship; Tetragastrin

The effects of iontophoretically applied cholecystokinin fragments and cholecystokinin antagonists on neurons of the dorsal lateral geniculate nucleus were investigated with extracellular recordings in rats anesthetized with urethane. The peptide cholecystokinin-8S, which has affinity for both cholecystokinin-A and -B receptors, altered the baseline firing as well as the responses to visual stimuli of about one half of the investigated neurons (90 out of 190). Excitatory effects predominated (P < 0.01, Wilcoxon test), although inhibitory effects were also observed. The effects of cholecystokinin-8S were dose-dependent. Neurons sensitive to cholecystokinin-8S could be found in all regions of the dorsal lateral geniculate nucleus, but they differed in their susceptibility to cholecystokinin in relation to their location. The B-agonist, BOC-cholecystokinin-4, also changed the baseline firing as well as the light-evoked activity of dorsal lateral geniculate nucleus neurons. The effects were either excitatory or inhibitory. Changes induced by cholecystokinin-8S could be effectively blocked by the cholecystokinin-B antagonist, CAM 1028 (19 out of 22 cholecystokinin-sensitive neurons tested). The cholecystokinin-A antagonist, Ge 410, blocked cholecystokinin-induced effects in 10 out of 16 neurons. These results indicate that the modulation of geniculate cell firing by cholecystokinin is mediated by both A-and B-receptor types.

Topics: Animals; Cholecystokinin; Geniculate Bodies; Iontophoresis; Male; Neurons; Rats; Rats, Wistar; Receptors, Cholecystokinin; Sincalide; Tetragastrin

The effects of the acutely administered cholecystokinin (CCK) agonists CCK tetrapeptide (BOC-CCK-4) and sulfated CCK octapeptide (CCK-8S) were examined in four animal models of anxiety in rats. In the elevated plus maze, BOC-CCK-4 reduced the time spent in the open arms and the number of entries into the open arms. BOC-CCK-4 but not the anorectic acting CCK-8S increased the suppression of feeding in a conflict paradigm based on novelty suppressed feeding in hungry rats. In the two-compartment black-and-white box, BOC-CCK-4 decreased the time spent and locomotor activity in the white compartment. In the ultrasound vocalization test, using rat pups separated from the mother, BOC-CCK-4 increased the number of distress calls. No evidence was found for inducing anxiety-like behaviour by CCK-8S.

Topics: Animals; Anxiety; Anxiety, Separation; Behavior, Animal; Conflict, Psychological; Feeding Behavior; Male; Motor Activity; Rats; Rats, Wistar; Sincalide; Tetragastrin; Vocalization, Animal

Replacement of Met31 by (N-Me)Nle in CCK8 or CCK4 has been shown to improve the affinity and selectivity for CCK-B receptors. In order to obtain molecules with enhanced bioavailability, two novel series of protected tetrapeptides of the general formula Boc-Trp30-X-Asp-Y33 have been developed. Introduction of (N-Me)Nle and the bulky, aromatic naphthylalaninamide (Nal-NH2) in positions X and Y, respectively, does not greatly modify the affinity for guinea pig brain CCK-B receptors. In contrast, incorporation of hindering N-methyl amino acids such as (N-Me)Phe, (N-Me)Phg, or (N-Me)Chg, but not their non-methylated counterparts, in position X induced a large decrease in affinity for the CCK-B binding sites. Among the various peptides synthesized, Boc-[(N-Me)Nle31,1Nal-NH2(33)]CCK4 (2) (KI = 2.8 nM), Boc-[Phg31,1Nal-NH2(33)]CCK4 (15) (KI = 14 nM), and Boc-[Phg31,1Nal-N(CH3)2(33)]CCK4 (17) (KI = 39 nM) displayed good affinities for brain CCK-B receptors and had good selectivity ratios. These pseudopeptides, in which the presence of unnatural and hydrophobic residues is expected to improve their penetration of the central nervous system, were shown to be very resistant to brain peptidases. Interestingly, whereas compounds 2 and 15 proved to be full agonists for rat hippocampal CCK-B receptors when measured in an electrophysiological assay, compound 17 behaved as a potent antagonist in the same test and displayed a good affinity in rat brain KI(CCK-B) = 51 nM as compared to the Merck antagonist L365,260,KI(CCK-B) = 12 nM. This illustrates a simple means to obtain CCK-B antagonists and suggests that the free, CONH2 group plays a critical role in the recognition of the agonist state of brain CCK-B receptors.

Topics: Amino Acid Sequence; Animals; Benzodiazepinones; Brain; Cholecystokinin; Guinea Pigs; Male; Molecular Sequence Data; Peptide Fragments; Phenylurea Compounds; Rats; Rats, Sprague-Dawley; Receptors, Cholecystokinin; Structure-Activity Relationship; Tetragastrin

Novel Boc-CCK-4 derivatives were communicated recently as having high potency and selectivity for the CCK-A receptor (Shiosaki et al. J. Med. Chem. 1990, 33, 2950-2952). While Boc-CCK-4 binds selectively to the CCK-B receptor, replacement of the methionine with an N epsilon-substituted lysine dramatically reversed receptor selectivity, leading to the development of this novel series of tetrapeptides. A detailed structure-activity analysis of a series of urea-substituted tetrapeptides, represented by the general structure Boc-Trp-Lys(N epsilon-CO-NHR)-Asp-Phe-NH2, revealed that a number of substituted phenyl, naphthyl, and aliphatic urea residues in the lysine side chain yielded potent and selective CCK-A ligands. These tetrapeptides elicit full agonist responses in stimulating pancreatic amylase release that are effectively blocked by a selective CCK-A receptor antagonist. Conversion of the urea to a thiourea significantly reduced CCK-A binding potency as did replacement of the lysine with the homologous ornithine or homolysine. Tetrapeptides that were partial agonists (less than 80% efficacy) in phosphoinositide (PI) hydrolysis relative to CCK-8 did not exhibit high-dose inhibition of amylase secretion in guinea pig acini.

Topics: Amino Acid Sequence; Amylases; Animals; Guinea Pigs; Hydrolysis; Molecular Sequence Data; Pancreas; Phosphatidylinositols; Receptors, Cholecystokinin; Tetragastrin; Urea

Cholecystokinin octapeptide (CCK-8) administered either systemically (IP) or centrally (ICV) suppresses several types of behavior in mice including exploratory locomotion, rearing and grooming. At doses equimolar to those active for CCK-8, neither desulfated CCK-8 (CCK-8-DS), nor the protected C-terminus tetrapeptide fragment, BOC-CCK-4, is behaviorally active when administered either centrally or systemically. A potent and selective antagonist to the peripheral type (Type A) CCK receptor, A-65186, when given systemically, blocked the effects of systemically administered CCK-8, but failed to block the effects of ICV administered CCK-8. Central administration of A-65186 blocked the effects of ICV administered CCK-8. These results demonstrate that administration of exogenous CCK-8 to mice can suppress exploratory locomotion by acting either centrally or peripherally and that in either case the demonstrated behavioral effects are mediated via a "peripheral" type (Type A) CCK receptor.

Topics: Animals; Dose-Response Relationship, Drug; Drug Interactions; Injections, Intraperitoneal; Injections, Intraventricular; Male; Mice; Motor Activity; Quinolines; Receptors, Cholecystokinin; Sincalide; Tetragastrin