gr-73632 and Gerbillinae

The neurokinin-1 (NK-1) antagonist LY686017 showed activity in preclinical anxiety models. The clinical development of LY686017 included a PET study and a proof-of-concept in social anxiety disorder (SAD). [(11)C]GR205171 was used healthy volunteers receiving 1-100mg/d LY686017 for 28 days to determine brain receptor occupancy (RO). The mean NK-1 RO increased ranged from 25% with 1mg to 93% with 100mg. Subsequently, a 12-week randomized clinical trial tested LY686017 vs. paroxetine, or placebo in SAD. Pharmacokinetic (PK)/RO modeling based on the PET results predicted that once daily dosing of >30mg LY686017 led to sustained trough RO of over 80%. 189 outpatients(1) suffering from SAD were randomly assigned to 12-weeks treatment with 50mg/d LY686017 (N=77), placebo (N=74), or 20mg/d paroxetine (N=38). There was no significant difference between LY686017 and placebo as measured with the Liebowitz Social Anxiety scale (LSAS). The active comparator paroxetine showed positive trends on primary and secondary measures. The plasma concentrations were above the level expected to produce maximal brain NK-1 RO based on the PK/RO relationship obtained in the human PET investigation. Thus, further evaluation of LY686017 for the treatment of SAD does not seem warranted.

Topics: Adult; Animals; Anti-Anxiety Agents; Anxiety Disorders; Brain; Dose-Response Relationship, Drug; Gerbillinae; Humans; Male; Neurokinin-1 Receptor Antagonists; Paroxetine; Peptide Fragments; Pyridines; Radioligand Assay; Radionuclide Imaging; Receptors, Neurokinin-1; Stereotyped Behavior; Substance P; Treatment Outcome; Triazoles

Experiments were conducted to develop a model to study the effect of oral and topical administration of the NK1 receptor antagonist aprepitant, on scratching behaviour in gerbils. The gerbil was selected due to its relevance for human NK1 receptor pharmacology. Intradermal injection of a specific NK1 receptor agonist GR73632 (100 nmol/100 µl) at the rostral back of gerbils produced scratching of the injection site. This could be attenuated by intradermal co-administration of a selective NK1 receptor antagonist aprepitant (30-100-300 nmol), demonstrating the role of dermal NK1 receptor in elicitation of scratching behaviour. Likewise, scratching was attenuated by oral (0.3-3-30 mg/kg) or topical application (0.01-0.1-1% w/v) of aprepitant and pharmacokinetic analysis of aprepitant levels in brain, blood and skin supported that efficacy of topically applied aprepitant was due to dermal rather than central target engagement. In conclusion, we showed that NK1 agonist-induced scratching in the gerbil can be reversed by systemic and topical administration of aprepitant. This test system may provide a useful model for the in vivo assessment of putative antipruritic agents.

Topics: Administration, Oral; Administration, Topical; Animals; Antipruritics; Aprepitant; Disease Models, Animal; Drug Evaluation, Preclinical; Gerbillinae; Humans; Injections, Intradermal; Morpholines; Neurokinin-1 Receptor Antagonists; Peptide Fragments; Pruritus; Receptors, Neurokinin-1; Substance P

Data suggest that substance P could play an important role in pruritus, and therefore, blockade of the neurokinin (NK)-1 receptor might be antipruritic. Thus, we explored in the Mongolian gerbil the effect on scratching behaviour, induced by intra-dermal injection of the NK-1 receptor-specific agonist GR73632, of oral administration of the NK-1 receptor antagonist orvepitant. Orvepitant at all doses tested (0.1-10 mg/kg p.o.) produced a profound inhibition of GR73632 (30 nmol i.d.) induced hindlimb scratching; the minimum effective dose of orvepitant in this model was identified as ≤0.1 mg/kg. The data generated supported the proposition that the antipruritic potential of orvepitant should be evaluated in clinical trials.

Topics: Administration, Oral; Animals; Antipruritics; Bridged Bicyclo Compounds, Heterocyclic; Disease Models, Animal; Dose-Response Relationship, Drug; Gerbillinae; Neurokinin-1 Receptor Antagonists; Peptide Fragments; Piperidines; Pruritus; Substance P

The pharmacological properties of the novel neurokinin-1 (NK(1)) receptor antagonist FK886, ([3,5-bis(trifluoromethyl)phenyl][(2R)-2-(3-hydroxy-4-methylbenzyl)-4-{2-[(2S)-2-(methoxymethyl)morpholin-4-yl]ethyl}piperazin-1-yl]methanone dihydrochloride), were studied. FK886 potently inhibited the binding of [(125)I]Bolton-Hunter-labeled substance P ([(125)I]BH-SP; 100 pM) to human NK(1) receptors expressed in Chinese hamster ovary (CHO) cells (IC(50)=0.70 nM). It also possessed high affinities for dog, ferret, gerbil and guinea pig NK(1) receptors, but not for rat NK(1) receptor. FK886 was highly selective for the NK(1) receptor, with 250- and >20000-fold selectivity for human NK(1) over NK(2) and NK(3), respectively. Further, it did not inhibit radioligand binding at 54 different sites, including receptors, ion channels and transporters. FK886 inhibited substance P (3.2 nM)-induced inositol phosphate formation in human NK(1) receptor-expressing CHO cells (IC(50)=1.4 nM) without stimulating NK(1) receptors. The antagonism exerted by FK886 against human NK(1) receptor was insurmountable in saturation binding experiments, with both the affinity and B(max) of [(125)I]BH-SP being significantly reduced. After intravenous administration, FK886 (0.01-0.1 mg/kg) dose-dependently inhibited the foot-tapping behavior induced by intracerebroventricular administration of a selective NK(1) receptor agonist, GR73632 (10 pmol), in gerbils, with significant inhibition being observed at doses of 0.032-0.1 mg/kg, indicating excellent brain penetration. The brain penetration of FK886 was further demonstrated by the cerebral distribution of radioactivity after intravenous injection of radiolabeled FK886. Taken together, these results demonstrate that FK886 is a potent, highly selective and centrally active, insurmountable antagonist of the NK(1) receptor, and suggest that FK886 antagonizes various NK(1) receptor-mediated biological effects in the central nervous system.

Topics: Animals; CHO Cells; Cricetinae; Cricetulus; Gerbillinae; Humans; Male; Morpholines; Neurokinin-1 Receptor Antagonists; Peptide Fragments; Piperazines; Rats; Rats, Sprague-Dawley; Receptors, Neurokinin-1; Recombinant Proteins; Substance P; Tissue Distribution

The pharmacological properties of T-2328 were evaluated as an antagonist of the tachykinin neurokinin 1 (NK(1)) receptor. T-2328 inhibited the specific binding of [(3)H][Sar(9),Met(O(2))(11)]substance P to tachykinin NK(1) receptors in human lymphoblastic IM9 cells with K(i) of 0.08 nM. In the same assay, K(i) for aprepitant, a brain-penetrating NK(1) antagonist, was 1.3 nM. The antagonism of T-2328 is highly selective for the human NK(1) receptors since the affinities for human NK(2), NK(3) receptors, and 13 other kinds of receptors and ion channels were >1000-fold lower than for NK(1) receptors. Reduction in Bmax with no change in affinity suggests the non-competitive nature of T-2328 interaction with the NK(1) receptor. T-2328 (0.03-0.1 mg/kg, i.v.) and aprepitant (1 - 3 mg/kg, i.v.) significantly prevented the GR73632 (i.c.v.)-induced foot tapping response in gerbils. The potencies of T-2328 in both in vitro and in vivo studies were more than 10 times greater than those of aprepitant. I.v. administration of T-2328 (0.1-0.3 mg/kg) potently blocked both acute and delayed emetic responses induced by cisplatin (5 mg/kg, i.p.) in ferrets. It is concluded that T-2328 is a potent, centrally active NK(1) antagonist. T-2328 may have potential as a novel therapeutic agent for the treatment of chemotherapy-induced emesis.

Topics: Animals; Antiemetics; Antineoplastic Agents; Aprepitant; Binding Sites; Brain; Cell Line, Tumor; CHO Cells; Cisplatin; Cricetinae; Cricetulus; Disease Models, Animal; Dose-Response Relationship, Drug; Ferrets; Gerbillinae; Humans; Kinetics; Male; Morpholines; Motor Activity; Neurokinin-1 Receptor Antagonists; Peptide Fragments; Piperidines; Receptors, Neurokinin-1; Substance P; Transfection; Vomiting

Intracerebroventricular (i.c.v.) administration of tachykinin NK(1) receptor agonists induces tapping of the hind legs in gerbils, so-called gerbil foot tapping, which is thought to reflect a fear-related response. The aim of the present study was to examine how ligands selective for NK(1), NK(2) and NK(3) receptors affect the gerbil foot tap response. Agonists selective for NK receptor subtypes were administered i.c.v. and the gerbil foot tap response was monitored. The effect of systemically administered antagonists was also studied. The interaction of ligands with gerbil NK(1) receptors was evaluated using autoradiography on gerbil brain slices with [(3)H]-Sar,Met(O(2))-substance P or [(3)H]GR205171 as radioligand. The effects of ligands on NK(1) and NK(3) receptor-mediated increases in intracellular calcium in vitro were studied in Chinese hamster ovary cells expressing the cloned gerbil receptors. The selective NK(1) receptor agonist ASMSP and the selective NK(3) receptor agonist senktide induced dose-dependent increases in gerbil foot tapping with similar potency. The maximal effect of senktide was approximately 40% of the maximal response evoked by ASMSP. The effects of ASMSP and senktide were blocked by administration of the selective NK(1) receptor antagonist CP99,994 (10 micromol/kg s.c.). The effects of senktide, but not ASMSP, were blocked by administration of the selective NK(3) receptor antagonist SB223412 (50 micromol/kg i.p.). Senktide did not displace NK(1) receptor radioligand binding and was >1000-fold less potent than ASMSP at activating gerbil NK(1) receptors. The selective NK(3) receptor agonist senktide evokes fear-related gerbil foot tapping, an effect which probably involves indirect enhancement of NK(1) receptor signalling.

Topics: Animals; Autoradiography; Behavior, Animal; Brain; Calcium; CHO Cells; Cloning, Molecular; Cricetinae; Cricetulus; Dose-Response Relationship, Drug; Foot; Gerbillinae; Injections, Intraventricular; Male; Neurokinin A; Neurokinin-1 Receptor Antagonists; Peptide Fragments; Piperidines; Quinolines; Receptors, Neurokinin-2; Receptors, Neurokinin-3; Substance P

The synthesis and structure-activity relations for a new class of centrally active NK-1 receptor antagonists are described. The new compounds are based on piperazine 2 and contain an oxime ether functionality. Several new compounds have high affinity for the NK-1 receptor and show good antagonistic activity in the gerbil foot-tapping assay.

Topics: Administration, Oral; Animals; Anti-Anxiety Agents; CHO Cells; Cricetinae; Drug Evaluation, Preclinical; Ethers; Gerbillinae; Humans; Molecular Conformation; Motor Activity; Neurokinin-1 Receptor Antagonists; Oximes; Peptide Fragments; Piperazines; Structure-Activity Relationship; Substance P

Identification of the selective neurokinin NK(1) receptor antagonist, 2-(R)-(1-(R)-3,5-Bis(trifluromethyl)phenylethoxy)-3-(S)-(4-fluoro)phenyl-4-(3-oxo-1,2,4-triazol-5yl)methylmor-phine (MK-869), as a novel therapeutic approach for anxiety/depression has led to increased use of the Mongolian gerbil in behavioural studies since the gerbil NK(1) receptor pharmacology is similar to human, but not rat or mouse. Within this species, foot tapping and immobility elicited by aversive conditioning, as well as social interaction have been shown to be sensitive to clinically used anxiolytic and antidepressant agents and also NK(1) receptor antagonists. The high levels of NK(1) receptor binding in the amygdala as well as preclinical studies demonstrating increased release of substance P and corresponding internalisation of NK(1) receptors in the basolateral amygdala in response to stressful stimuli suggest that the BLA may represent a potential site of action for NK(1) receptor antagonists in anxiety and/or depression. Therefore, in the current study, we assessed the effect of bilateral BLA lesions in male Mongolian gerbils on footshock-induced foot tapping and immobility, social interaction, and NK(1)-agonist-induced foot tapping. Lesioned gerbils exhibited reduced immobility time during fear conditioning, a non-significant reduction in immobility time when re-exposed to the conditioned stimulus (CS) 24 h later, and increased social interaction in the gerbil social interaction task. In contrast, BLA lesions had no effect on NK(1)-agonist-induced foot tapping. These data provide further support that the gerbil BLA is a potential site for NK(1) receptor antagonists to attenuate anxiety-related behaviours.

Topics: Amygdala; Analysis of Variance; Animals; Aprepitant; Behavior, Animal; Conditioning, Classical; Dose-Response Relationship, Drug; Drug Interactions; Fear; Gerbillinae; Ibotenic Acid; Immobility Response, Tonic; Interpersonal Relations; Male; Morpholines; Motor Activity; Neurokinin-1 Receptor Antagonists; Peptide Fragments; Substance P

The ability of the substance P (NK(1) receptor) antagonist (SPA) L-760735 to inhibit conditioned fear was assessed in gerbils using a four plate apparatus. Animals that had been treated with diazepam (3 mg/kg) or L-760735 (3 mg/kg) 30 min before a 3 min conditioning session in the apparatus exhibited a release of plate crossings during the retest session approximately 3 h later. Plate crossings were also increased when animals received diazepam or L-760735 30 min before the retest session. In contrast, fluoxetine and venlafaxine (30 mg/kg) did not exhibit anxiolytic-like effects. During the retest session, gerbils drummed their hind feet on the floor; this behaviour was not observed spontaneously in gerbils that were naïve to the apparatus. Foot drumming was abolished by pretreatment with L-760735 or diazepam (3 mg/kg) but was markedly increased following administration of fluoxetine or venlafaxine (30 mg/kg). Foot drumming elicited by aversive conditioning alone or in combination with fluoxetine was abolished by administration of L-760735 and by amygdala lesions involving the basolateral and lateral nuclei, indicating that this behaviour is an alarm signal or fear response mediated via release of substance P in brain circuits involving the amygdala. The observations provide further evidence for an anxiolytic-like profile of SPAs in preclinical assays and demonstrate a clear difference between the actions of SPAs and established antidepressant drugs.

Topics: Amygdala; Animals; Anti-Anxiety Agents; Conditioning, Psychological; Fear; Female; Gerbillinae; Ibotenic Acid; Male; Morpholines; Neurokinin-1 Receptor Antagonists; Peptide Fragments; Substance P

The ability of tachykinin NK1 receptor antagonists to inhibit GR73632 (D-Ala-[L-Pro9,Me-Leu8]substance P-(7-11))-induced foot tapping in gerbils was employed as an indirect measure of brain penetration and this was compared with their ability to prevent acute emesis induced by cisplatin in ferrets. (+)-GR203040 ((2S,3S and 2R,3R)-2-methoxy-5-tetrazol-1-yl-benzyl-(2-phenyl-piperidin- 3-yl)-amine), CP-99,994 ((2S,3S)-cis-3-(2-methoxybenzylamino)-2-phenyl piperidine) dihydrochloride), and L-742,694 (2-(S)-(3,5-bis(trifluoromethyl)benzyloxy)-3-(S)-phenyl-4-(5-(3-oxo-1,2, 4-triazolo)methylmorpholine) potently inhibited GR73632-induced foot tapping (ID50 < or = 0.85 mg/kg), and acute retching induced by cisplatin (ID50 < or = 0.18 mg/kg). RPR100893 ((3aS,4S,7aS)-7,7-diphenyl-4-(2-methoxyphenyl)-2-[(S)-2-(2-m ethoxyphenyl)proprionyl] perhydroisoindol-4-ol) was not a potent antagonist of retching (ID50 4.1 mg/kg) or foot tapping (ID50 > 10 mg/kg). High doses (3-10 mg/kg) of CGP49823 ((2R,4S)-2-benzyl-1-(3,5-dimethylbenzoyl)-N-[(4-quinolinyl)methyl] -4-piperineamine) dihydrochloride), FK888 (N2-[(4R)-4-hydroxy-1-(1-methyl-1H-indol-3-yl)carbonyl-L-propyl]-N-methy l-N-phenylmethyl-L-3-(2-naphthyl)-alaninamide), and LY303870 ((R)-1-[N-(2-methoxybenzyl)acetylamino]-3-(1H-indol-3-yl)-2-[N-(2-(4-(pi peridinyl)piperidin-1-yl)acetyl)amino]propane) were required to inhibit foot tapping; these agents were not anti-emetic in this dose range. SR140333 ((S)-1-[2-[3-(3,4-dichlorphenyl)-1 (3-isopropoxyphenylacetyl)piperidin-3-yl] ethyl]-4-phenyl-1 azaniabicyclo [2.2.2]octane; 3-10 mg/kg) failed to inhibit foot tapping or emesis. Affinities for the human and ferret tachykinin NK1 receptor were highly correlated (r = 0.93, P = 0.0008). Inhibition of foot tapping in gerbils, but not NK1 receptor binding affinity, predicted anti-emetic activity in ferrets (r = 0.75, P < 0.01). These findings confirm that the anti-emetic activity of tachykinin NK1 receptor antagonists is dependent on brain penetration.

Topics: Administration, Oral; Animals; Antiemetics; Brain; CHO Cells; Cisplatin; Cloning, Molecular; Cricetinae; Ferrets; Gerbillinae; Humans; Infusions, Intravenous; Membranes; Neurokinin-1 Receptor Antagonists; Peptide Fragments; Radioligand Assay; Substance P

1. Stimulation of sensory nerves causes release of tachykinins, including substance P (SP) and neurokinin A (NKA), which produce a variety of respiratory effects via NK-1 and NK-2 receptors, respectively. Hence, development of a compound which could potently and equivalently antagonize both receptors was pursued. 2. MDL 105,172A ((R)-1-[3-(3,4-dicholorophenyl)-1-(3,4,5-trimethoxybenzoyl)- 3-pyrrolidinyl]-4- phenyl-piperidine-4-morpholinecarboxamide) exhibited high affinity for NK-1 (4.34 nM) and NK-2 (2.05 nM) receptors. In vitro, the compound antagonized SP (pA2 = 8.36) or NKA (pA2 = 8.61)-induced inositol phosphate accumulation in tachykinin monoreceptor cell lines. 3. In anaesthetized guinea-pigs, MDL 105,172A inhibited SP-induced plasma protein extravasation (ED50 = 1 mg kg-1, i.v.) and [beta-Ala8]NKA 4-10-induced bronchoconstriction (ED50 = 0.5 mg kg-1, i.v.) indicating NK-1 and NK-2 antagonism, respectively. 4. Capsaicin was used to elicit respiratory effects in anaesthetized and conscious guinea-pigs; the latter were inhibited by MDL 105,172A following i.v. (ED50 = 1 mg kg-1) or oral (ED50 = 20 mg kg-1) administration. Hence, MDL 105,172A can inhibit pulmonary responses to tachykinins released endogenously in the airways. 5. At doses up to 200 mg kg-1, p.o., MDL 105,172A failed to inhibit repetitive hind paw tapping induced by i.c.v GR 73632, and NK-1 selective agonist, in gerbils, whereas CP-99,994 (0.87 mg kg-1, s.c.) completely ablated the effect. These data suggest that MDL 105,172A does not penetrate the central nervous system (CNS) and its tachykinin antagonism is restricted to the periphery. 6. MDL 105,172A is a non-peptide, potent, equivalent antagonist of NK-1 and NK-2 receptors. Its ability to inhibit both exogenously administered as well as endogenously released tachykinins support its use in examining the role of sensory neuropeptides in diseases associated with neurogenic inflammation including asthma.

Topics: Animals; Behavior, Animal; Capillary Permeability; Gerbillinae; Guinea Pigs; In Vitro Techniques; Inositol Phosphates; Male; Morpholines; Neurokinin A; Neurokinin-1 Receptor Antagonists; Peptide Fragments; Piperidines; Plethysmography, Whole Body; Radioligand Assay; Receptors, Neurokinin-2; Respiratory Mechanics; Substance P; Tachykinins

Intravenous administration of the NK1 receptor antagonist L-733,060 to gerbils 3 h before intraplantar injection of formalin caused a dose-dependent and complete inhibition of the late, but not early, phase nociceptive response (paw licking). The ID50 for L-733,060 (0.17 mg/kg) revealed a greater than 50-fold separation in potency over its less active enantiomer L-733,061 (ID50 > or = 10 mg/kg). In contrast, the non-brain penetrant quaternary ketone NK1 receptor antagonist, L-743,310 (3 mg/kg), did not attenuate the response to formalin, indicating that the antinociceptive effect of blockade of NK1 receptors by L-733,060 in this assay is centrally-mediated. These findings add to the preclinical evidence that NK1 receptor antagonists may be of therapeutic use as centrally-acting analgesics.

Topics: Animals; Female; Foot; Formaldehyde; Gerbillinae; Injections; Male; Motor Activity; Neurokinin-1 Receptor Antagonists; Nociceptors; Peptide Fragments; Piperidines; Stereoisomerism; Substance P; Time Factors

The in vivo pharmacological profiles of the selective tachykinin NK1 receptor agonists, [Sar9,Met(O2)11]substance P and GR 73632, were examined in gerbils. Both agonists induced a pronounced chromodacryorrhea following intravenous injection which was stereoselectively antagonised by the tachykinin NK1 receptor antagonist, CP-99,994, but not by its inactive enantiomer, CP-100,263, or the rat-selective tachykinin NK1 receptor antagonist, RP 67,580. In contrast, chromodacryorrhea was not observed following intravenous injection of the selective tachykinin NK2 receptor agonist, [beta-Ala8]neurokinin A-(4-10), or the selective tachykinin NK3 receptor agonist, senktide. These results suggest that [Sar9,Met(O2)11]substance P-induced chromodacryorrhea results from activation of peripheral tachykinin NK1 receptors. Repetitive hind paw tapping was also observed in gerbils but only following intracerebroventricular injection of [Sar9,Met(O2)11]substance P or GR 73632. Furthermore, GR 73632-induced hind paw tapping was significantly attenuated by co-administration of the peptide tachykinin NK1 receptor antagonist, GR 82334, or intravenous injection of CP-99,994. Thus, in contrast to chromodacryorrhea, repetitive hind paw tapping may result from activation of central tachykinin NK1 receptors.

Topics: Animals; Behavior, Animal; Dose-Response Relationship, Drug; Drug Synergism; Gerbillinae; Harderian Gland; Indoles; Injections, Intravenous; Injections, Intraventricular; Isoindoles; Male; Neurokinin A; Peptide Fragments; Piperidines; Receptors, Neurokinin-1; Substance P

The inhibition of GR73632-induced foot tapping in gerbils by central nervous system (CNS) penetrant of tachykinin NK1 receptor antagonists was investigated. Intracerebroventricular infusion of the highly selective tachykinin NK1 receptor agonist GR73632 (3 pmol) induced a vigorous repetitive hind foot tapping response which was inhibited by CP-99,994 (ID50 = 0.06 mg/kg i.v.) but not by its less active enantiomer, CP100,263 (10 mg/kg i.v.). Similarly, the poorly CNS penetrant quaternised compound, L-743,310, failed to inhibit foot tapping at doses up to 3 mg/kg i.v. In contrast, all three compounds inhibited chromodacryorrhoea induced by systemic administration of GR73632 (0.5 nmol i.v.) (ID50 = 0.06, 2.95 and 0.004 mg/kg i.v., respectively). These findings confirm that foot tapping and chromodacryorrhoea in gerbils provide simple assays for the central and peripheral activation of tachykinin NK1 receptors.

Topics: Analysis of Variance; Animals; Behavior, Animal; Biological Assay; Blood-Brain Barrier; Female; Gerbillinae; Injections, Intravenous; Injections, Intraventricular; Male; Neurokinin-1 Receptor Antagonists; Peptide Fragments; Substance P; Tears