bluuebunny/crossref_metadata_2025_split huggingface dataset:
Pharmacological characterization of the bradykinin B2 receptor: inter‐species variability and dissociation between binding and functional responses
Authors: Paquet J ‐L , Luccarini J ‐M , Fouchet C , Defrêne E , Loillier B , Robert C , Bélichard P , Cremers B , Pruneau D 
Published: January 2005

The present study addresses the differences in binding profiles and functional properties of the human and rat bradykinin (BK) B2 receptor using various kinin receptor peptide derivatives as well as the non‐peptide receptor antagonists WIN 64338 (phosphonium, [[4‐[[2‐[[bis(cyclohexylamino)methylene]amino]‐3‐(2‐naphtalenyl)1‐oxopropyl]amino]‐phenyl]‐methyl]tributyl, chloride, monohydro‐chloride), and FR173657 (E)‐3‐(6‐acetamido‐3‐pyridyl)‐N‐[‐N‐[2,4‐dichloro‐3‐[(2‐methyl‐8‐quinolinyl)oxymethyl]‐phenyl]N‐methylamino carbonyl methyl] acrylamide.
[3H]‐BK bound with a similar affinity to membranes of Chinese hamster ovary cells (CHO‐K1) expressing the cloned human (hB2‐CHO) or rat (rB2‐CHO) B2 receptor, human embryonic intestine cells (INT407) expressing the native B2 receptor, human umbilical vein (HUV) and rat uterus (RU). WIN 64338 and FR173657 bound with a 3.8–6.6 fold and 7.0–16.3 fold higher affinity the rat than the human B2 receptor, respectively. The affinity values of BK derivatives as well as non‐peptide antagonists were reduced by 6–23 fold in physiological HBSS compared to low ionic strength TES binding buffer.
BK (0.01–3000 nM) increased inositol triphosphates (IP3) levels in hB2‐CHO, rB2‐CHO and INT407 cells. The B2 receptor antagonist, Hoe 140 (D‐Arg0‐[ Hyp3, Thi5, D‐Tic7, Oic8]‐BK) at 10−7 M, significantly shifted to the right the IP3 response curves to BK giving apparent pKB values of 8.56, 9.79 and 8.84 for hB2‐CHO, rB2‐CHO and INT407 cells, respectively.
In human isolated umbilical vein, Hoe 140, D‐Arg0‐[Hyp3, D‐Phe7, Leu8]‐BK and NPC 567 had a lower potency in functional assays (pKB 8.18, 5.77 and 5.60, respectively) than expected from their affinity in binding studies (pKi 10.52, 8.64 and 8.27, respectively).
FR173657 behaved as a high affinity ligand with pKi values of 8.59 and 9.81 and potent competitive antagonist with pKB values of 7.80 and 8.17 in HUV and RU, respectively. FR173657 bound with a similar affinity the cloned and native bradykinin B2 receptor in human (pKi of 8.66 and 8.59, respectively) and in rat (pKi 9.67 and 9.81, respectively).
In conclusion, we suggest that the binding buffer composition has to be taken into account when screening new compounds and that inter‐species differences should be considered when setting up animal models with the aim of developing bradykinin B2 receptor antagonists as therapeutic agents.

British Journal of Pharmacology (1999) 126, 1083–1090; doi:10.1038/sj.bjp.0702403
Pro‐ and anti‐inflammatory actions of thrombin: a distinct role for proteinase‐activated receptor‐1 (PAR1)
Authors: Vergnolle Nathalie , Hollenberg Morley D , Wallace John L 
Published: January 2005

Thrombin has well characterized pro‐inflammatory actions that have recently been suggested to occur via activation of its receptor, proteinase‐activated receptor‐1 (PAR1).
In the present study, we have compared the effects of thrombin to those of two peptides that selectively activate the PAR1 receptor, in a rat hindpaw oedema model. We have also examined whether or not thrombin can exert anti‐inflammatory activity in this model.
Both thrombin and the two PAR1 activating peptides induced significant oedema in the rat hindpaw following subplantar injection.
The oedema induced by thrombin was abolished by pre‐incubation with hirudin, and was markedly reduced in rats in which mast cells were depleted through treatment with compound 48/80 and in rats pretreated with indomethacin. In contrast, administration of the PAR1 activating peptides produced an oedema response that was not reduced by indomethacin and was only slightly reduced in rats pretreated with compound 48/80.
Co‐administration of thrombin together with a PAR1 activating receptor resulted in a significantly smaller oedema response than that seen with the PAR1 activating peptide alone. This anti‐inflammatory effect of thrombin was abolished by pre‐incubation with hirudin.
These results demonstrate that the pro‐inflammatory effects of thrombin occur through a mast‐cell dependent mechanism that is, at least in part, independent of activation of the PAR1 receptor. Moreover, thrombin is able to exert anti‐inflammatory effects that are also unrelated to the activation of PAR1.

British Journal of Pharmacology (1999) 126, 1262–1268; doi:10.1038/sj.bjp.0702408
Evidence for in vitro expression of B1 receptor in the mouse trachea and urinary bladder
Authors: Trevisani Marcello , Schmidlin Fabien , Tognetto Michele , Nijkamp Frans P , Gies Jean‐Pierre , Frossard Nelly , Amadesi Silvia , Folkerts Gert , Geppetti Pierangelo 
Published: January 2005

Motor responses to des‐Arg9‐bradykinin and bradykinin were studied in the isolated mouse trachea (precontracted with carbachol, 10 μM) and the urinary bladder of either Swiss, C57Bl/6J or bradykinin B2 receptor knockout (Bk2r−/−) mice after 1–6 h in vitro. The expression of mRNA for the mouse B1 receptor in tracheal and urinary bladder tissues was also studied by using Northern blot analysis.
In isolated tracheae, des‐Arg9‐bradykinin produced a relaxant response that increased over time: no response was observed after 1 h of incubation, whereas after 6 h the maximum response (1 μM) was 68–84% of the relaxation produced by isoproterenol (1 μM) in the three mouse strains. The relaxant response to bradykinin (1 μM) observed at 1 h (38–51% of isoproterenol) was increased (62–65% of isoproterenol) after 6 h in Swiss and C57Bl/6J mice, but was absent in Bk2r−/− mice. In the presence of cycloheximide, des‐Arg9‐bradykinin did not cause any response at 6 h.
Similar findings were obtained in the urinary bladder: at 1 h des‐Arg9‐bradykinin (1 μM) did not cause any motor effect, whereas at 6 h it caused a contraction that was 28–59% of that produced by carbachol (1 μM) in the three mouse strains. Cycloheximide blocked the response to des‐Arg9‐bradykinin. Bradykinin (1 μM) contracted urinary bladders at 1 h (34–35% of carbachol), as well as at 6 h (66–77% of carbachol) in Swiss and C57Bl/6J strains, but was without effect in Bk2r−/− mice.
Northern blot hybridization with a specific cDNA probe against mouse B1 receptor mRNA using total RNA extracted from tracheae and urinary bladders freshly removed from Swiss and Bk2r−/− mice revealed minimal expression. However, marked hybridization was detected 150 min after in vitro exposure in both tissues.
Evidence is provided that in vitro exposure of mouse trachea and urinary bladder causes a time‐dependent induction of B1 receptors that cause relaxation and contraction, respectively.

British Journal of Pharmacology (1999) 126, 1293–1300; doi:10.1038/sj.bjp.0702410
Increased dopaminergic and 5‐hydroxytryptaminergic activities in male rat brain following long‐term treatment with anabolic androgenic steroids
Authors: Thiblin Ingemar , Finn Anja , Ross Svante B , Stenfors Carina 
Published: January 2005

The effects of treating groups of rats with four different anabolic androgenic steroids (AAS) (testosterone, nandrolone, methandrostenolone, and oxymetholone) on 5‐hydroxytryptamine (5‐HT) and dopamine (DA) neurones in different brain regions were examined. The AAS was injected six times with 1 week's interval and the rats were sacrificed 2 days after the final injection. 5‐HT and its metabolite 5‐hydroxyindoleacetic acid (5‐HIAA), DA and its metabolites 3,4‐dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) were measured. The effect on DA and 5‐HT synthesis rate was analysed as the accumulation of 3,4‐dihydroxyphenyl‐alanine (DOPA) and 5‐hydroxytryptophan (5‐HTP), respectively, after inhibition of the amino acid decarboxylase with NSD‐1015 (3‐hydroxy‐benzylhydrazine dihydrochloride). Additionally, the monoamine oxidase (MAO) activity was analysed in the hypothalamus.
The DOPAC+HVA/DA ratio was increased in the striatum in all treatment groups. However, the synthesis rate of DA was significantly increased only in the methandrostenolone treated group.
The 5‐HIAA/5‐HT ratio was increased in all treatment groups in the hippocampus, in the frontal cortex in the methandrostenolone‐treated animals and in the hypothalamus in the testosterone‐ and oxymetholone‐treated rats, while the 5‐HT synthesis rate was not affected by the AAS‐treatments.
The MAO‐A activity was increased in the oxymetholone‐treated rats while the other treatment groups were unaffected. The MAO‐B activity was not changed.
The results indicate that relatively high doses of AAS increase dopaminergic and 5‐hydroxytryptaminergic metabolism in male rat brain, probably due to enhanced turnover in these monaminergic systems.

British Journal of Pharmacology (1999) 126, 1301–1306; doi:10.1038/sj.bjp.0702412
Effects of short chain alkanols on the inducible nitric oxide synthase in a glial cell line
Authors: Syapin Peter J , Rendon Alexia , Huron David R , Militante Julius D 
Published: January 2005

Ethanol inhibits inducible nitric oxide synthase (iNOS) expression in C6 glioma cells by an unknown mechanism. Because relatively high concentrations are needed for inhibition in drug‐naïve cells (IC50∼150 mM), suppression due to cytotoxicity is one possible mechanism that has not been ruled out. Therefore, the present study examined the effects of ethanol and other alkanols on C6 glioma cell viability and iNOS activity to better understand the mechanism for inhibition.
iNOS expression was induced in cell culture with lipopolysaccharide and phorbol ester treatment. Nitrite accumulation in culture medium, the in vitro conversion of [3H]‐L‐arginine to [3H]‐L‐citrulline, and immunoblotting were used to quantify iNOS induction and activity. Trypan blue exclusion, extracellular release of lactate dehydrogenase, and quantity of total cell protein were used as measures of viability.
Short chain alkanols, methanol through 1‐heptanol, concentration‐dependently inhibited nitrite accumulation. Longer chain alkanols, 1‐octanol and 1‐decanol, did not except at cytotoxic concentrations. Experiments indicated short chain alkanol inhibition was not due to direct actions on iNOS catalytic activity, but that it transpires during iNOS induction. Immunoblots showed reduced iNOS protein levels.
Correlation analysis ruled out iNOS inhibition as being due to decreased cell number, total cell protein, or cell viability. In contrast, there was significant correlation with physical measures of lipophilicity.
In conclusion, inhibition of iNOS expression by ethanol and other short chain alkanols is not due to cytotoxicity. Instead, the strong correlation with lipophilicity suggests the inhibition derives from an interaction with unknown hydrophobic cellular sites.

British Journal of Pharmacology (1999) 126, 1253–1261; doi:10.1038/sj.bjp.0702417
Pharmacological activities of TEI‐8362, a novel inhibitor of human neutrophil elastase
Authors: Mitsuhashi Hiroaki , Nonaka Takashi , Hamamura Ichiro , Kishimoto Tadashi , Muratani Emiko , Fujii Katsuhiko 
Published: January 2005

TEI‐8362, 4‐(N‐(3‐((3‐carboxypropyl)amino)‐8‐methyl‐1‐oxo‐4‐azaisochromen‐6‐yl)carbamoyl)‐4‐((phenylmethoxy)carbonylamino)butanoic acid (C26H28N4O9) is a novel inhibitor of human neutrophil elastase (HNE). We evaluated its pharmacological profile in vitro and in vivo.
TEI‐8362 demonstrated potent inhibition of HNE with a Ki value of 1.38×10−9 M. Its selectivity for HNE among a variety of proteases ranged from 163 fold to 68,000 fold in favour of HNE.
The pulmonary haemorrhage that occurred after i.t. instillation of HNE to hamsters was inhibited by either i.t., i.v., or inhalant administration of TEI‐8362.
Intratracheal administration of lipopolysaccharide induced pulmonary neutrophilia. Twenty‐four hours after lipopolysaccharide administration, the additional treatment with formyl‐methionyl‐leucyl‐phenylalanine resulted in a specific neutrophil‐dependent acute lung injury. In this model, lung injury was significantly attenuated by i.t., i.v., or inhalant administration of TEI‐8362.
These pharmacological actions of TEI‐8362 suggest that this drug has therapeutic value in the treatment of destructive lung diseases due to neutrophils.

British Journal of Pharmacology (1999) 126, 1147–1152; doi:10.1038/sj.bjp.0702425
Prostaglandin endoperoxide‐dependent vasospasm in bovine coronary arteries after nitration of prostacyclin synthase
Authors: Zou Minghui , Jendral Michelle , Ullrich Volker 
Published: January 2005

In the present study we used a bioassay to study the effects of peroxynitrite (ONOO−) on angiotensin II (A‐II)‐triggered tension in isolated bovine coronary arteries in order to show the consequences of the previously reported PGI2‐synthase inhibition by ONOO− in this model. The following results were obtained:

1 μmol L−1 ONOO− impaired A‐II‐induced vasorelaxation and caused a second long lasting constriction phase. Indomethacin (10−5M) prevented both effects. U51605, a dual blocker of PGI2‐synthase and thromboxane (TX)A2‐synthase mimicked the effects of ONOO−.
The selective TXA2/prostaglandin endoperoxide (PGH2) receptor antagonist SQ29548 antagonized the second vasoconstriction phase after ONOO−‐treatment. Since a generation of TXA2 and 8‐iso‐prostaglandin F2α could be excluded a direct action of unmetabolized PGH2 on the TXA2/PGH2 receptor was postulated.
ONOO− dose‐dependently inhibited the conversion of 14C‐PGH2 into 6‐keto‐PGF1α in isolated bovine coronary arteries with an IC50‐value of 100 nM.
Immunoprecipitation of 3‐nitrotyrosine‐containing proteins with a monoclonal antibody revealed PGI2‐synthase as the only nitrated protein in bovine coronary arteries treated with 1 μmol  l−1 ONOO−.
Using immunohistochemistry a co‐localization of PGI2‐synthase and nitrotyrosine‐containing proteins was clearly visible in both endothelial and vascular smooth muscle cells. We concluded that ONOO− not only eliminated the vasodilatory, growth‐inhibiting, antithrombotic and antiadhesive effects of PGI2 but also allowed and promoted an action of the potent vasoconstrictor, prothrombotic agent, growth promoter, and leukocyte adherer, PGH2.

British Journal of Pharmacology (1999) 126, 1283–1292; doi:10.1038/sj.bjp.0702434
Mechanisms of melatonin‐induced vasoconstriction in the rat tail artery: a paradigm of weak vasoconstriction
Authors: Lew Michael J , Flanders Samantha 
Published: January 2005

Vasoconstrictor effects of melatonin were examined in isolated rat tail arteries mounted either in an isometric myograph or as cannulated pressurized segments. Melatonin failed by itself to mediate observable responses but preactivation of the arteries with vasopressin (AVP) reliably uncovered vasoconstriction responses to melatonin with maxima about 50% of maximum contraction. Further experiments were conducted with AVP preactivation to 5–10% of the maximum contraction.
Responses to melatonin consisted of steady contractions with superimposed oscillations which were large and irregular in isometric but small in isobaric preparations. Nifedipine (0.3 μM) reduced the responses and abolished the oscillations. Charybdotoxin (30 nM) increased the magnitude of the oscillations with no change in the maximum response.
Forskolin (0.6 μM) pretreatment increased the responses to melatonin compared to control and sodium nitroprusside (1 μM) treated tissues. The AVP concentration required for preactivation was 10 fold higher than control in both the forskolin and nitroprusside treated groups.
In isometrically‐mounted arteries treated with nifedipine, melatonin receptor agonists had the potency order 2‐iodomelatonin>melatonin>S20098>GR196429, and the MT2‐selective antagonist luzindole antagonized the effects of melatonin with a low pKB of 6.1±0.1.
It is concluded that melatonin elicits contraction of the rat tail artery via an mt1 or mt1‐like receptor that couples via inhibition of adenylate cyclase and opening of L‐type calcium channels. Calcium channels and charybdotoxin‐sensitive K+ channels may be recruited into the responses via myogenic activation rather than being coupled directly to the melatonin receptors.
It is proposed that the requirement of preactivation for overt vasoconstrictor responses to melatonin results from the low effector reserve of the melatonin receptors together with the tail artery having threshold inertia. Potentiative interactions between melatonin and other vasoconstrictor stimuli probably also result from the threshold inertia. A simple model is presented and a general framework for consideration of interactions between weak vasoconstrictor agonists and other vasoconstrictor stimuli is discussed.

British Journal of Pharmacology (1999) 126, 1408–1418; doi:10.1038/sj.bjp.0702435
Characterization of receptors for calcitonin gene‐related peptide and adrenomedullin on the guinea‐pig vas deferens
Authors: Poyner David R , Taylor Gillian M , Tomlinson A Elaine , Richardson Alan G , Smith David M 
Published: January 2005

The receptors which mediate the effects of calcitonin gene‐related peptide (CGRP), amylin and adrenomedullin on the guinea‐pig vas deferens have been investigated.
All three peptides cause concentration dependant inhibitions of the electrically stimulated twitch response (pD2s for CGRP, amylin and adrenomedullin of 7.90±0.11, 7.70±0.19 and 7.25±0.10 respectively).
CGRP8–37 (1 μM) and AC187 (10 μM) showed little antagonist activity against adrenomedullin.
Adrenomedullin22–52 by itself inhibited the electrically stimulated contractions of the vas deferens and also antagonized the responses to CGRP, amylin and adrenomedullin.
[125I]‐adrenomedullin labelled a single population of binding sites in vas deferens membranes with a pIC50 of 8.91 and a capacity of 643  fmol  mg−1. Its selectivity profile was adrenomedullin> AC187>CGRP=amylin. It was clearly distinct from a site labelled by [125I]‐CGRP (pIC50=8.73, capacity=114  fmol  mg−1, selectivity CGRP>amylin=AC187>adrenomedullin). [125I]‐amylin bound to two sites with a total capacity of 882 fmol mg−1.
Although CGRP has been shown to act at a CGRP2 receptor on the vas deferens with low sensitivity to CGRP8–37, this antagonist displaced [125I]‐CGRP with high affinity from vas deferens membranes. This affinity was unaltered by increasing the temperature from 4°C to 25°C, suggesting the anomalous behaviour of CGRP8–37 is not due to temperature differences between binding and functional assays.

British Journal of Pharmacology (1999) 126, 1276–1282; doi:10.1038/sj.bjp.0702437
A non‐pungent triprenyl phenol of fungal origin, scutigeral, stimulates rat dorsal root ganglion neurons via interaction at vanilloid receptors
Authors: Szallasi A , Bíró T , Szabó T , Modarres S , Petersen M , Klusch A , Blumberg P M , Krause J E , Sterner O 
Published: January 2005

A [3H]‐resiniferatoxin (RTX) binding assay utilizing rat spinal cord membranes was employed to identify novel vanilloids in a collection of natural products of fungal origin. Of the five active compounds found (scutigeral, acetyl‐scutigeral, ovinal, neogrifolin, and methyl‐neogrifolin), scutigeral (Ki=19 μM), isolated from the edible mushroom Albatrellus ovinus, was selected for further characterization.
Scutigeral induced a dose‐dependent 45Ca uptake by rat dorsal root ganglion neurons with an EC50 of 1.6 μM, which was fully inhibited by the competitive vanilloid receptor antagonist capsazepine (IC50=5.2 μM).
[3H]‐RTX binding isotherms were shifted by scutigeral (10–80 μM) in a competitive manner. The Schild plot of the data had a slope of 0.8 and gave an apparent Kd estimate for scutigeral of 32 μM.
Although in the above assays scutigeral mimicked capsaicin, it was not pungent on the human tongue up to a dose of 100 nmol per tongue, nor did it provoke protective wiping movements in the rat (up to 100 μM) upon intraocular instillation.
In accord with being non‐pungent, scutigeral (5 μM) did not elicit a measurable inward current in isolated rat dorsal root ganglion neurons under voltage‐clamp conditions. It did, however, reduce the proportion of neurons (from 61 to 15%) that responded to a subsequent capsaicin (1 μM) challenge. In these neurons, scutigeral both delayed (from 27 to 72 s) and diminished (from 5.0 to 1.9 nA) the maximal current evoked by capsaicin.
In conclusion, scutigeral and its congeners form a new chemical class of vanilloids, the triprenyl phenols. Scutigeral promises to be a novel chemical lead for the development of orally active, non‐pungent vanilloids.

British Journal of Pharmacology (1999) 126, 1351–1358; doi:10.1038/sj.bjp.0702440
Characterization of the electrophysiological and pharmacological effects of 4‐iodo‐2,6‐diisopropylphenol, a propofol analogue devoid of sedative‐anaesthetic properties
Authors: Sanna Enrico , Motzo Costantino , Usala Marcello , Serra Mariangela , Dazzi Laura , Maciocco Elisabetta , Trapani Giuseppe , Latrofa Andrea , Liso Gaetano , Biggio Giovanni 
Published: January 2005

Several derivatives and analogues of the general anaesthetic 2,6‐diisopropylphenol (propofol) have been recently synthesised with the aim of exploring the structure‐activity relationships.
In the present study, the effects of one such compound, 4‐iodo‐2,6‐diisopropylphenol (4‐I‐Pro), on γ‐aminobutyric acid type A (GABAA) receptors in vitro were compared with its in vivo effects in rodents. Human GABAA receptors were expressed in Xenopus oocytes, and the actions of 4‐I‐Pro on receptor function were compared with those of propofol by two‐electrode voltage‐clamp recording.
Similar to propofol, 4‐I‐Pro directly activated Cl− currents in the absence of GABA at all combinations of receptor subunits tested. However, the efficacy of 4‐I‐Pro in inducing direct activation of α1β2γ2S receptors was markedly less than that of propofol.
Similarly to propofol, 4‐I‐Pro potentiated in a concentration‐dependent manner GABA‐evoked Cl− currents measured at different GABAA receptor constructs.
As expected, intraperitoneal injection of propofol induced sedation, ataxia, and loss of the righting reflex in rats. In contrast, administration of 4‐I‐Pro failed to produce any of these behavioural effects.
Administration of 4‐I‐Pro to rats reduced in a dose‐dependent manner the incidence of tonic‐clonic seizures induced by pentylenetetrazol and induced an anticonflict effect as measured in the Vogel test.
Microdialysis revealed that, like propofol, administration of 4‐I‐Pro reduced acetylcholine release in the hippocampus of freely moving rats.
These results demonstrate that para‐substitution of the phenol ring of propofol with iodine yields a compound that exhibits anticonvulsant and anticonflict effects, but is devoid of sedative‐hypnotic and anaesthetic properties. Thus, 4‐I‐Pro possesses pharmacological characteristics more similar to anxiolytic and anticonvulsant drugs than to general anaesthetics.

British Journal of Pharmacology (1999) 126, 1444–1454; doi:10.1038/sj.bjp.0702449