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The objective was to explore the epileptogenic effects of several drugs specific to the antihistaminic H1 receptor on NMDA and GABA responses. With patch-clamp whole-cell recordings of hippocampal neurons of rats, we observe the effects of several histaminergic agonists and antagonists on the NMDA and GABA currents. During the NMDA rundown and GABA rundown, we applied the drugs and observe the effect on the responses, and compare them to control conditions. Mepyramine with nanomolar concentrations increases significantly the NMDA responses about 35 %, this effect is mimicked by another Anti H1 drug Triprolidine. Histamine alone has no effect on NMDA rundown but 100 microM Histamine can reduce partially the effect of Mepyramine. An H1 receptor agonist 2,3 Bromophenyl Histamine also shows the same properties as Histamine. The H1 receptor of Histamine seems to be crucial during these experiments, blockade of its constitutive activity by Mepyramine induces a significant increase of the NMDA responses. On the GABA response, the same nanomolar dose of Mepyramine has no effect on the rundown and we also observed no effect of Histamine on the GABA rundown. An interaction between Histamine H1 receptor and the NMDA is revelated by using Mepyramine in nanomolar concentration, Histamine has no effect on these responses also in these experiments Mepyramine acts as an inverse agonist blocking the constitutive activity of the H1 receptor. The constitutive activity of the H1 receptor seems to be crucial in the regulation of NMDA receptor activities.

The histamine H3 receptor was identified in the 80's by our group as a presynaptic autoreceptor inhibiting histamine synthesis and release in the rat brain. Sixteen years later, cloning of the related human H3 receptor revealed the existence of isoforms, species pharmacological differences and a high constitutive (spontaneous) activity of the receptor. All these molecular findings have to be taken into account for optimizing aimed at clinical applications ligands. H3 receptor inverse agonists, by increasing histamine neuron activity, promote arousal and enhance cognitive performances. Pharmaceutical firms have shown considerable interest for this new class of drugs and many programmes of clinical development of H3 receptor inverse agonists for the treatment of arousal and cognitive disorders are presently being conducted.

Starting from thioperamide, the first potent and selective H 3 -receptor histamine antagonist, analogues have been synthesized and tested in vitro on rat cerebral cortex to explore structure-activity relationships. The aim has been to design potent compounds which do not possess the thiourea group of thioperamide and which may have improved brain penetration. In a short series of open chain thiourea analogues, the optimum chain length for H 3 -antagonist potency was found to be (CH 2 ) 3 . Compounds derived from histamine and possessing an aromatic nitrogen-containing heterocycle on the side chain amino group in place of thiourea show H 3 -antagonist activity. Furthermore, when the heterocycle is 2-pyridyl, electron-withdrawing substituents (e.g. NO 2 , CF 3 , CO 2 Me) in the pyridine 5-position increased potency. The synthesis of 4-[4(5)-imidazolyl]piperidine and its conversion into the (trifluoromethyl)pyridyl analogue 5b of thioperamide is described ; however, 5b is not as potent as thioperamide. Replacing imidazole by pyridine or substituting imidazole on the remote N considerably reduced potency. Replacing the side-chain NH by S increased potency still further and the most potent compound is 2-{[2-[4(5)-imidazolyl]ethyl]thio}-5-nitropyridine (UCL 1199) which has K i = 4.8 nM

Summary In search for potential histamine H3-receptor agonists a series of mono- and dialkyl-substituted histamine derivatives was synthesized. All target compounds were tested in vitro for their agonist activity at H3-, H2-, and H1-receptors. Introduction of one ethyl or two methyl residues into histamine led to compounds with decreased histamine H3-agonist potency in most cases. However, the non-chiral α,α-dimethylhistamine (15) was identified to be three times as active as histamine itself at H3-receptors. In addition 15 shows high receptor selectivity being 20 000 times as active at H3- as at H2- and H1-receptors, respectively. (αR)-α,Nτ-Dimethylhistamine 23, which is a potential metabolite of (αR)-α-methylhistamine 1, proved to be inactive at all three histamine receptor subtypes.

BACKGROUND AND PURPOSE The histamine H(3) receptor was identified as the autoreceptor of brain histaminergic neurons. After its cloning, functional H(3) receptor isoforms generated by a deletion in the third intracellular loop were found in the brain. Here, we determined if this autoreceptor was the long or the short isoform. EXPERIMENTAL APPROACH We hypothesized that the deletion would affect H(3) receptor stereoselectivity. The effects of the enantiomers of two chiral ligands, N(α)-methyl-α-chloromethylhistamine (N(α) Me-αClMeHA) and sopromidine, were investigated on cAMP formation at the H(3(445)) and H(3(413)) receptor isoforms, common to all species. They were further compared with their effects at autoreceptors. They were also compared on [(35)S]GTPγ[S] binding to membranes of rat cerebral cortex, striatum and hypothalamus, the richest area in autoreceptors. KEY RESULTS The stereoselectivity of N(α) Me-αClMeHA enantiomers as agonists was similar at the H(3(413)) receptor isoform and autoreceptors, but lower at the long isoform. While (S) sopromidine did not discriminate between the isoforms, (R) sopromidine was an antagonist at the H(3(413)) receptor isoform and autoreceptors, but a full agonist at the long isoform. In rat brain, stereoselectivity of N(α) Me-αClMeHA was higher in the hypothalamus than in cerebral cortex or striatum, whereas the opposite pattern was found for sopromidine. CONCLUSIONS AND IMPLICATIONS The pharmacological profiles of H(3) receptor isoforms differed markedly, showing that the function of autoreceptors was fulfilled by a short isoform, such as the H(3(413)) receptor. Development of drugs selectively targeting autoreceptors might enhance their therapeutic efficacy and/or decrease incidence of side effects.

Summary In search for potential histamine H3-receptor agonists a series of mono- and dialkyl-substituted histamine derivatives was synthesized. All target compounds were tested in vitro for their agonist activity at H3-, H2-, and H1-receptors. Introduction of one ethyl or two methyl residues into histamine led to compounds with decreased histamine H3-agonist potency in most cases. However, the non-chiral α,α-dimethylhistamine (15) was identified to be three times as active as histamine itself at H3-receptors. In addition 15 shows high receptor selectivity being 20 000 times as active at H3- as at H2- and H1-receptors, respectively. (αR)-α,Nτ-Dimethylhistamine 23, which is a potential metabolite of (αR)-α-methylhistamine 1, proved to be inactive at all three histamine receptor subtypes.

Starting from thioperamide, the first potent and selective H 3 -receptor histamine antagonist, analogues have been synthesized and tested in vitro on rat cerebral cortex to explore structure-activity relationships. The aim has been to design potent compounds which do not possess the thiourea group of thioperamide and which may have improved brain penetration. In a short series of open chain thiourea analogues, the optimum chain length for H 3 -antagonist potency was found to be (CH 2 ) 3 . Compounds derived from histamine and possessing an aromatic nitrogen-containing heterocycle on the side chain amino group in place of thiourea show H 3 -antagonist activity. Furthermore, when the heterocycle is 2-pyridyl, electron-withdrawing substituents (e.g. NO 2 , CF 3 , CO 2 Me) in the pyridine 5-position increased potency. The synthesis of 4-[4(5)-imidazolyl]piperidine and its conversion into the (trifluoromethyl)pyridyl analogue 5b of thioperamide is described ; however, 5b is not as potent as thioperamide. Replacing imidazole by pyridine or substituting imidazole on the remote N considerably reduced potency. Replacing the side-chain NH by S increased potency still further and the most potent compound is 2-{[2-[4(5)-imidazolyl]ethyl]thio}-5-nitropyridine (UCL 1199) which has K i = 4.8 nM

In search for novel leads for histamine H3-receptor antagonists a number of amine derivatives of different (1 H-imidazol-4-yl)anilines and omega-(1 H-imidazol-4-yl)alkanamines were prepared. Pharmacological in vitro H3-receptor investigations of the prepared urea, amide, inverse amide, thioamide, and thiourea derivatives on synaptosomes of rat cerebral cortex proved that the aniline derivatives are inactive at H3-receptors, whereas derivatives of the omega-(1 H-imidazol-4-yl)-alkanamines showed moderate to good activity. Some compounds are active in the nanomolar concentration range. The most potent compounds in this series are the thioamide derivative 7 and the urea derivatives 11, 12 of 3-(1 H-imidazol-4-yl) propanamine. Therefore, the urea derivatives were tested in vitro on isolated organs of the guinea pig for their activity on the other two histamine receptor subtypes proving their high selectivity. In vivo studies of the effects of the urea derivatives 11 and 12 on brain Nt-methylhistamine levels, a test of central H3-receptor activity after peroral application to mice, showed no detectable activity. Thus, the urea type antagonists are useful potent and selective H3-receptor tools for in vitro studies and for investigations of ligand-receptor interactions.

Novel ω-phenyl substituted and unsubstituted alkyl and alkenyl imidazole derivatives were prepared and tested for their antagonist activity in vitro and in vivo at histamine H 3 -receptors. Some compounds showed high in vitro and in vivo H 3 -receptor activity despite their structure bearing no polar moiety in the centre of the molecule which is a common structural feature of all other antagonists known. Quite probably there are further in vivo effects for some compounds resulting from other receptor interactions.

The histamine H3 receptor was identified in the 80's by our group as a presynaptic autoreceptor inhibiting histamine synthesis and release in the rat brain. Sixteen years later, cloning of the related human H3 receptor revealed the existence of isoforms, species pharmacological differences and a high constitutive (spontaneous) activity of the receptor. All these molecular findings have to be taken into account for optimizing aimed at clinical applications ligands. H3 receptor inverse agonists, by increasing histamine neuron activity, promote arousal and enhance cognitive performances. Pharmaceutical firms have shown considerable interest for this new class of drugs and many programmes of clinical development of H3 receptor inverse agonists for the treatment of arousal and cognitive disorders are presently being conducted.

With the recent development of new hybrid compounds having histamine H3 receptor antagonist with combined histamine Ntau-methyltransferase (HMT) inhibitory potency an innovative approach was described in the research of novel lead compounds modulating histaminergic neurotransmission. Several compounds containing an ether moiety derived from the recently published 4-(3-piperidinopropoxy)phenylaminoquinoline derivatives (like FUB 836), were synthesized in this study and tested for their affinity at cloned human histamine H3 (hH3) receptors and on the inhibition of rat HMT. Besides different heterocycles, e.g. nitro- or amino-substituted pyridines, quinolines, benzothiazole or pyrroline, three classes of compounds were produced: heteroaromatic 3-piperidinopropyl ethers, keto- or imino-substituted 4-(3-piperidinopropyl)phenyl ethers and 4-(3-piperidinopropyl)phenyl ethers with substituted (alkyl)aminopyridines. Whereas the (3-piperidinopropoxy)heterocycles showed only moderate activities on both test models, the 4-(3-piperidinopropoxy)phenyl derivatives were identified as potent histamine H3 receptor ligands and/or HMT inhibitors. Ki values up to 0.42 nM were found for the affinity to the hH3 receptor. HMT inhibitory potency was identified with IC50 values about 0.3 microM for the most potent compounds in this series. Comparison of the pyridine-containing derivatives to recently published quinoline analogues showed a decrease in potencies for the pyridines. The dual activity, H3 receptor affinity and HMT inhibition, was moderate to good. For all compounds affinities at hH3 receptors were higher than their inhibitory HMT potencies. The described new histamine H3 receptor antagonists with an ether moiety represent a further promising step in our investigations for a dual activity.

The histamine H3 receptor has been identified in the rat brain as a presynaptic autoreceptor inhibiting histamine synthesis and release. Following its recent cloning, more than fifteen years later, the existence of isoforms, species pharmacological differences and the high constitutive activity of the receptor were established. All these molecular findings have to be taken into account for the optimization of the ligands aiming at a clinical use. They show the interest of antagonists/inverse agonists in the symptomatic treatment of schizophrenia and cognitive and attentional deficits.

Novel derivatives of 3-(1H-imidazol-4-yl)propanol were designed on the basis of lead compounds belonging to the carbamate or ether series possessing (partial) agonist properties on screening assays of the histamine H(3) receptor. One pair of enantiomers in the series of alpha-methyl-branched chiral carbamates was stereoselectively prepared in high optical yields. Enantiomeric purity was checked by Mosher amide derivatives of precursors and capillary electrophoresis of the final compounds with trimethyl-beta-cyclodextrin as chiral selector, and was determined to be/=95%. The novel compounds were investigated in various histamine H(3) receptor assays in vitro and in vivo. Some compounds displayed partial agonist activity on synaptosomes of rat brain cortex, whereas others exhibited antagonist properties only. Selected compounds were investigated in [(125)I]iodoproxyfan binding studies on the human histamine H(3) receptor and showed high affinity in the nanomolar concentration range. Under in vivo conditions after oral administration to mice, some of the compounds exhibited partial or full agonist activity in the brain at low dosages. The (S)-enantiomer of one pair of chiral carbamates (9) proved to be the eutomer; thus, the (S)-enantiomer was selected for further pharmacological studies. In a peripheral in vivo test model in rats, measuring the level of inhibition of capsaicin-induced plasma extravasation, (S)-9 again proved its high oral agonist potency with full intrinsic activity (ED(50) values of 0.07-0.1 mg/kg depending on tissue).

In search of novel ways to enhance histaminergic neurotransmission in the central nervous system, a new class of nonimidazole histamine H(3) receptor ligands were developed that simultaneously possess strong inhibitory activity on the main histamine metabolizing enzyme, histamine N-methyltransferase (HMT). The novel compounds contain an aminoquinoline moiety, which is an important structural feature for HMT inhibitory activity, connected by different spacers to a piperidino group (for H(3) receptor antagonism). Variation of the spacer structure provides two different series of compounds. One series, having only an alkylene spacer between the basic centers, led to highly potent HMT inhibitors with moderate to high affinity at human histamine H(3) receptors. The second series possesses a p-phenoxypropyl spacer, which may be extended by another alkylene chain. This latter series also showed strong inhibitory activity on HMT, and in most cases, the H(3) receptor affinity even surpassed that of the first series. One of the most potent compounds with this dual mode of action is 4-(4-(3-piperidinopropoxy)phenylamino)quinoline (34) (hH(3), K(i) = 0.09 nM; HMT, IC(50) = 51 nM). This class of compounds showed high antagonist potency and good H(3) receptor selectivity in functional assays in guinea pig on H(1), H(2), and H(3) receptors. Because of low or missing in vivo activity of two selected compounds, the proof of concept of these valuable pharmacological tools for the supposed superior overall enhancing effect on histaminergic neurotransmission failed to appear hitherto.

The hypoxia-inducible factor (HIF-1alpha), a basic helix-loop-helix transcription factor, is known to heterodimerize with ARNT1, a nuclear translocator, to trigger the overexpression in many cells of genes involved in resistance to hypoxia. Although HIF-1alpha and ARNT1 are both expressed in brain, their cellular localization and function therein are unknown. Here, using in situ hybridization and immunocytochemistry, we show that HIF-1alpha is expressed in normoxic cerebral neurons together with not only ARNT1 but also ARNT2, a cerebral translocator homologous to ARNT1 but displaying, unlike ARNT1, a selective neuronal expression. In contrast, other potential partners of the translocators, i.e. the aryl hydrocarbon receptor (AHR) and the single-minded protein 2 (SIM2), are not expressed in the adult brain. We also identify two splice variants of HIF-1alpha in brain, one of which dimerizes with ARNT2 even more avidly than with ARNT1. The resulting heterodimer, in contrast with the HIF-1alpha/ARNT1 complex, does not recognize the HIF-1-binding site of the hypoxia-induced erythropoietin (Epo) gene, suggesting that it controls transcription of a distinct set of genes. We therefore propose that HIF-1alpha and ARNT2 function as preferential dimerization partners in neurons to control specific responses, some of which may not be triggered by hypoxia. In support of this proposal, in nonhypoxic PC12 cells constitutively coexpressing HIF-1alpha, ARNT1 and ARNT2, downregulation of either HIF-1alpha or ARNT2, obtained with selective antisense nucleotides, resulted in inhibition of [3H]thymidine incorporation.

The processes responsible for the limited ability to divide and long survival of neurons are not well understood but may involve aryl hydrocarbon receptor nuclear translocator 2 (ARNT2), a recently identified protein, apparently belonging to the basic helix-loop-helix superfamily of transcription factors, which is expressed almost exclusively in brain during the whole lifetime. In agreement, we show, in the rat, that ARNT2 immunoreactivity could be observed only within nuclei of brain neurons and of dividing and neuronal PC12 cells, a localization consistent with a role in transcription regulation. Cell death elicited either by focal ischaemia in brain or oxidative stress in PC12 cells was largely preceded by an almost complete suppression of ARNT2 expression. In contrast, when PC12 cell cycle progression was impaired, ARNT2 expression was enhanced. Finally, the downregulation of ARNT2 levels induced by antisense oligonucleotides prevented PC12 cell proliferation and induced apoptosis. These observations support the hypothesis that ARNT2 is a neuronal transcription factor, regulating cell cycle progression and preventing cell death, whose sustained expression might ensure brain neuron survival.

Histamine has been converted into a non-imidazole H3-receptor histamine antagonist by addition of a 4-phenylbutyl group at the N alpha-position followed by removal of the imidazole ring. The resulting compound, N-ethyl-N-(4-phenylbutyl)amine, remarkably has a Ki = 1.3 microM as an H3 antagonist. Using this as a lead compound, a novel series of homologous O and S isosteric tertiary amines was synthesised and structure-activity studies furnished N-(5-phenoxypentyl)pyrrolidine (Ki = 0.18 +/- 0.10 microM, for [3H]histamine release from rat cerebral cortex synaptosomes) which, more importantly, was active in vivo. Substitution of NO2 into the para position of the phenoxy group gave N-(5-p-nitrophenoxypentyl)pyrrolidine, UCL 1972 (Ki = 39 +/- 11 nM), ED50 = 1.1 +/- 0.6 mg/kg per os in mice on brain tele-methylhistamine levels.

Novel ω-phenyl substituted and unsubstituted alkyl and alkenyl imidazole derivatives were prepared and tested for their antagonist activity in vitro and in vivo at histamine H 3 -receptors. Some compounds showed high in vitro and in vivo H 3 -receptor activity despite their structure bearing no polar moiety in the centre of the molecule which is a common structural feature of all other antagonists known. Quite probably there are further in vivo effects for some compounds resulting from other receptor interactions.

A series of 4-(omega-(arylalkyloxy)alkyl)-1H-imidazoles and related sulphur-containing compounds have been prepared and evaluated for their histamine H3-autoreceptor antagonist in vitro potency in an assay on synaptosomes of rat cerebral cortex. In addition, the in vivo potency has been determined from the changes in N tau-methylhistamine levels in brain after p.o. administration to mice. Compounds with different alkyl chains and various aryl moities have been synthesized and tested to explore structure-activity relationships. Within this series of novel antagonists, (1H-imid-azol-4-yl)methyl and 2-(1H-imidazol-4-yl)ethyl ether derivatives showed low to moderate H3-receptor antagonist potency, whereas the corresponding allyl and propyl derivatives were compounds with high antagonist in vitro potency. Corresponding thioether or sulphoxide derivatives also showed antagonists activity. Additionally, some ether derivatives possessed high in vivo potency as well. The most active ether derivatives under in vivo conditions were 4-(3-(3-(4-fluorophenyl)propyloxy)propyl)-1H-imidazole (11b) and the corresponding chloro compound 11c (FUB 181) with ED50 values of 0.76 and 0.80 mg/kg, respectively. On the other hand, all compounds tested showed weak activity at histamine H1 or H2 receptors. Furthermore, the most promising ether FUB 181 exhibited low activity at adrenergic alpha 1, beta 1/2, serotonergic 5-HT2A, 5-HT3, and muscarinic M3 receptors. Time-course investigations of FUB 181 in mice showed a rapid mode of action with the highest value 3 h after p.o. application. Thus, FUB 181 appears to block histamine H3 receptors potently and selectively.

This study was performed in order to develop new leads for antagonists of the histamine H3-receptor subtype. omega-(1 H-Imidazol-4-yl)alkyl derivatives with ester, ketone or alcohol functionality in the side chain were synthesized and tested concerning their H3-receptor antagonist activity on synaptosomes of rat cerebral cortex. The novel compounds, which possess no nitrogen-containing polar group in the side chain of the imidazole moiety, presented moderate to high antagonist potency in vitro. In this series 3-(1 H-imidazol-4-yl)propyl-3-cyclopentylpropanoate (4) was the most potent compound in vitro with -log Ki = 8.5. Unfortunately, no central antagonist H3-receptor activity was detectable for ester derivatives in the in vitro H3-receptor assay based upon measurement of brain N tau-methylhistamine levels after p.o. administration to mice. Some of these novel antagonists are useful tools for investigations on ligand-receptor interaction because of their distinct receptor activities. On the other hand, the ketone derivative 1-(1 H-imidazol-4-yl)-7-phenyl-4-heptanone (9) in vitro presented an ED50-value of 3.5 +/- 1.5 mg/kg p.o. thus proving to be a new lead for further drug investigations. The most potent compounds in vitro and in vivo also showed high H3-receptor selectivity when tested at other histamine receptor subtypes.

In search for novel leads for histamine H3-receptor antagonists a number of amine derivatives of different (1 H-imidazol-4-yl)anilines and omega-(1 H-imidazol-4-yl)alkanamines were prepared. Pharmacological in vitro H3-receptor investigations of the prepared urea, amide, inverse amide, thioamide, and thiourea derivatives on synaptosomes of rat cerebral cortex proved that the aniline derivatives are inactive at H3-receptors, whereas derivatives of the omega-(1 H-imidazol-4-yl)-alkanamines showed moderate to good activity. Some compounds are active in the nanomolar concentration range. The most potent compounds in this series are the thioamide derivative 7 and the urea derivatives 11, 12 of 3-(1 H-imidazol-4-yl) propanamine. Therefore, the urea derivatives were tested in vitro on isolated organs of the guinea pig for their activity on the other two histamine receptor subtypes proving their high selectivity. In vivo studies of the effects of the urea derivatives 11 and 12 on brain Nt-methylhistamine levels, a test of central H3-receptor activity after peroral application to mice, showed no detectable activity. Thus, the urea type antagonists are useful potent and selective H3-receptor tools for in vitro studies and for investigations of ligand-receptor interactions.

5-Lipoxygenase-activating protein (FLAP) is an 18-kDa integral membrane protein required, in peripheral cells, for the activation of 5-lipoxygenase (5-LO) and for the resulting synthesis of leukotrienes from arachidonic acid. In the brain, the leukotrienes have been implicated in several pathophysiological events and in the electrophysiological effect of somatostatin, yet the cellular origin and role of these messenger molecules are still poorly understood. In the present study, we used reverse transcriptase-polymerase chain reaction, in situ hybridization, and immunohistochemistry to demonstrate that 5-LO and FLAP are expressed in various regions of the rat brain, including hippocampus, cerebellum, primary olfactory cortex, superficial neocortex, thalamus, hypothalamus, and brainstem. Highest levels of expression were observed in cerebellum and hippocampus. In the latter we demonstrate the colocalization of 5-LO and FLAP in CA1 pyramidal neurons. Moreover, electrophysiological experiments show that selective inhibition of FLAP with the compound MK-886 (0.25-1 microM) prevents the somatostatin-induced augmentation of the hippocampal K+ M-current. Our results provide necessary evidence for the presence and signaling role of 5-LO and FLAP in central neurons and strongly support their proposed participation in somatostatin-receptor transmembrane signaling.

Starting from thioperamide, the first potent and selective H3-receptor histamine antagonist, analogues have been synthesised and tested in vitro on rat cerebral cortex to explore structure-activity relationships. The aim has been to design compounds which do not possess the thiourea group of thioperamide and which may have improved brain penetration. Some compounds derived from histamine and possessing an aromatic nitrogen-containing heterocycle on the side-chain amino group show strong H3-antagonist activity. These have served as leads to provide aryloxyethyl- and aryloxy-propylimidazoles which are potent H3 antagonists of histamine. Structure-activity studies of the very potent and selective agonist, imetit (S-[2-imidazol-4-yl)ethyl]isothiourea), have explored the the transition between agonist, partial agonist and antagonist. The isosteric isourea is also a potent agonist. N,N'-Dibutyl-[S-[3-(imidazol-4-yl)propyl]isothiourea is a very potent antagonist having Ki = 1.5 nM.

By using a strategy based on nucleotide sequence homology, we have cloned an intronless DNA encoding a new putative member of G protein-coupled receptors. The deduced amino acid sequence of the rat OL1 receptor, together with its expression at high levels in a small subset of cells in the olfactory neuroepithelium indicate that OL1 is related to the recently discovered olfactory multigene family. PCR and in situ hybridization analyses showed the OL1 transcripts to be not only expressed in the olfactory epithelium, but also in the heart. This unexpected cardiac expression was developmentally regulated, being maximal at early postnatal stages and hardly detectable at adult stages. Moreover, this observation was not restricted to OL1 since it was extended to other putative olfactory receptors. Although its functional significance remains unknown, this transient cardiac expression suggests that receptors belonging to the olfactory superfamily, could be not only involved in odor coding, but also in cardiac morphogenesis. Another olfactory-specific gene transcript encoding PTP NE-3, a recently cloned receptor-type protein-tyrosine phosphatase, could also be detected in heart. The very low levels of expression observed in rat embryo and at early postnatal stages as compared to adult stages, suggest that protein-tyrosine phosphatases, as well as protein-tyrosine kinases, may play a role in the control of cardiac cell growth and morphogenetic processes.

Histamine, a ubiquitous cell-to-cell messenger, exerts its numerous actions through interaction with three pharmacologically distinct receptor subtypes, termed H1, H2 and H3. The design of selective agonists and antagonists has allowed to establish their respective pharmacological profile. Radioligand binding studies and, very recently, molecular biological studies have shown that they all belong to the superfamily of G-protein coupled receptors. H1 and H2-receptor antagonists have been successfully used for a long time in the treatment of allergy and ulcer, respectively. Some of them have been designed as highly potent and selective radioligands and have allowed to analyze the precise distribution of H1 and H2 receptors in various tissues including the brain. Recently, H1- and H2-receptor genes have been cloned in various animal species. Transfection of mammalian cells with these intronless genes has confirmed the respective coupling of H1 and H2 receptors with phospholipase C and adenylylcyclase. However, other known or unknown intracellular signals, could also be triggered by the stimulation in a transfected cell of a single H1 or H2 receptor through coupling to different G-proteins. A third histamine receptor subtype, the H3 receptor was evidenced in rodent and human brain by the inhibition of histamine release and synthesis it mediates in various areas. Thus, H3 receptors were considered as autoreceptors localized on histaminergic terminals. With the design of several potent and selective H3-receptor agonists and of an antagonist thioperamide, the critical role of H3 receptors in the control of histaminergic neurons in vivo was established.(ABSTRACT TRUNCATED AT 250 WORDS)

The effects of a new agonist of histamine (HA) H3 receptors, Imetit (S-[2-(4-(imidazolyl)ethyl]isothiourea) were investigated in vitro and in vivo and compared to those of (R)-alpha-methylhistamine [(R)-alpha-MeHA], a prototypic drug. Imetit inhibited the binding of [3H](R-alpha-MeHA to rat brain membranes with a Ki value of 0.1 +/- 0.01 nM. The release of endogenously synthesized [3H]HA induced by K(+)-depolarization from rat brain slices and synaptosomes was inhibited by Imetit with EC50 values of 1.0 +/- 0.3 and 2.8 +/- 0.7 nM, respectively. Imetit behaved as a full agonist and was about 4 times more potent than (R)-alpha-MeHA and 60 times more potent than HA. Thioperamide, a selective H3 receptor antagonist, elicited a parallel rightward shift of the concentration-response curve for Imetit with an apparent Ki value of 5.6 +/- 1.4 nM. Imetit potencies relative to HA were less than 0.1% and only 0.6% at HA H1 and H2 receptor reference systems, respectively. Imetit was found not to be a substrate or an inhibitor of HMT. After p.o. administration to mice or rats, Imetit decreased (by approximately 50%) the tele-MeHA level in the cerebral cortex with ED50 values of 1.0 +/- 0.3 and 1.6 +/- 0.3 mg/kg, respectively. This effect was still maximal after 6 hr. The in vivo potency and duration of action of Imetit were in the same range as those of (R)-alpha-MeHA. It is therefore concluded that Imetit represents a new potent and selective HA H3 receptor agonist.

Author(s): Ruat, M; Traiffort, E; Leurs, R; Arrang, JM; Kouyoumdjian, G; Tardivel-Lacombe, J; Piomelli, D; Diaz, G; Schwartz, JC

Among a series of methylated histamine derivatives, (R) alpha, (S) beta,-dimethylhistamine has been identified as a novel potent and selective H3-receptor agonist, even slightly more potent than (R) alpha-methylhistamine and displaying similar properties in vivo. Several studies suggesting that (R) alpha-methylhistamine might find various clinical applications, it was submitted to toxicological studies and initial clinical trials. The drug seems to display a very low animal toxicity, and, in humans, is well absorbed orally and metabolized via ring methylation before urinary excretion.

Several alkyl-substituted histamine derivatives have been synthesized and investigated for their agonistic potency on three classes of histamine receptors. While all investigated compounds are full agonists at H3-receptors their relative potency vs. histamine is strongly varying and culminates in alpha R, beta S-dimethylhistamine which is a highly potent and selective H3-receptor agonist.

Together with the two previous volumes of the "Handbook of Experimental Pharmacology" on histamine and antihistamines (volume 18, part 1, 1966 and part 2, 1978), the present volume yields a picture of a still rapidly developing field of research. It covers the process in histamine research during the past two decades. As a result of a consensus meeting in Munich in December 1988, a panel of eminent specialists arrived at common recommendations as to the usefulness of the available histamine assays for the most common experimental biomedical conditions. New information is presented about the mechanism of exocytosis. The old question about the role of histamine in the mechanism of gastric secretion and in cardiovascular and respiratory functions has been studied with new techniques, and the role of H1 and H2 receptors discussed. New observations have been made on the occurence and possible functions of histaminergic neurons and histamine receptors in CNS where a new type of receptor, the H3, seems to be widely represented. The structure and structure-activity relationships of H1 and H2 receptor antagonists are extensively treated. The indirect effects of histamine mediated via neuropeptides are discussed. Another topic covered is the presence of histamine in blood and food. New facts about the pathophysiology of histamine in anesthesia and surgery as well as in the pathogenesis of duodenal ulcer and other gastro-intestinal lesions are presented. Chapters are also devoted to new knowledge about the enzymatic formation and breakdown of histamine and about the distribution of the involved enzymes. The book should be useful for clinicians, practitioners, and researchers working in this exciting field. This monograph on pharmacology/ toxicology, physiology, immunology, dermatology is intended for clinicians and researchers.

In 25 cases of germinal testicular tumours of the adult, a technique of indirect immunoperoxydase was applied to histological sections with the aim of demonstrating a substance reacting with an anti-beta HCG antibody (sub-unit of chorionic gonadotrophin hormone). If the experimental conditions were appropriate, the substance should actually be beta HCG.

The recent development of technics of specific fixation using radio isotope-labelled tracer elements permits the demonstration of various types of receptors at the level of the lymphocyte membrane. Thus a new route is open in the understanding of the biochemical mechanisms controlling the physiology of the lymphocyte and its role in the immune response. Furthermore these lymphocyte receptors may represent to some degree, the state of the rarely accessible tissue receptors in human pathology. However, at present, the practical interest is limited by the absence of standardisation of technics and the heterogeneity of athe lymphocyte population studied.

In brain, histamine (HA) is the transmitter of a neuronal system resembling other monoaminergic systems. It is also present in mast cells from which it may control vascular and inflammatory processes. Its various actions are mediated not only by the two well known H1 and H2-receptor subclasses but also by the recently discovered H1-receptors, with distinct localization and pharmacology. H1-receptors mediate a series of biochemical responses which have several features in common: they require intact cells to be observed and largely depend upon the availability of Ca2+. H1 receptor-mediated responses include glycogenolysis, stimulation of cyclic GMP formation, potentiation of cyclic AMP formation. Recent studies indicate that H1-receptors are linked with phosphatidylinositol breakdown and generation of two intracellular signals which both contribute to the final response (e.g. in the cyclic AMP generation). H2-receptors seem.to be directly linked with an adenylate cyclase and their stimulation results in enhanced electrophysiologically recorded responses to excitatory agents. Finally whereas H1 and H2-receptors appear to be postsynaptically located, a novel subclass (H3) of HA receptors was recently revealed with a presynaptic localization. H3-receptors are autoreceptors mediating inhibition of HA release from and biosynthesis in histaminergic nerve terminals in the CNS. The physiological and pharmacological implications of three distinct receptor subclasses for HA will be discussed.

A simple derivative of histamine, alpha-methylhistamine i.e. 4-(2-aminopropyl)-imidazole, was shown to potently inhibit the K+-induced release of [3H] histamine from slices of rat cerebral cortex previously incubated in the presence of [3H] histidine. The maximal inhibition elicited by alpha-methylhistamine was of about 60% i.e. similar to that elicited by exogenous histamine. The effect occurred with an EC50 value of 4.3 +/- 1.1 X 10(-9) M about 10 times lower than that of histamine and was reversed by a H3-receptor antagonist. Since alpha-methylhistamine is known to display negligible potency at H1- and H2-receptors, this compound appears to be the first highly potent and selective H3-receptor agonist to be identified.

In contrast to cerebral histamine H1 and H2-receptors, histamine H3-receptors are presynaptically located on histamine-synthesizing nerve terminals (autoreceptors) and control the synthesis and release of the amine in cerebral neurons. Two imidazole derivatives were designed to interact at H3-receptors: (R) alpha-methylhistamine (alpha-MeHA), a chiral agonist, and thioperamide, a competitive antagonist derived from imidazolyl piperidine, both display high selectivity and potency at nanomolar concentrations in vitro. (R) alpha-MeHA, being about 15 times as potent as histamine itself, constitutes, when tritiated, a suitable probe for the radioassay of H3-receptors. Outside the brain, H3-receptor sites could be detected in the lung. The availability of new ligands made it possible to assess in vivo the physiological role of central and peripheral H3-receptors. The agonist and the antagonist modified in opposite directions histamine synthesis in the lung as in the brain, confirming the presence of H3-receptors in this peripheral organ. A large part of lung histamine being present within mast-cells, it is likely that these cells are endowed with H3-receptors controlling the amine synthesis and, possibly, release. Therefore, the novel agents may be of great practical interest in the field of allergy and inflammation.