Investigating the TNP-OVA and direct popliteal lymph node assays for the detection of immunostimulation by drugs associated with anaphylaxis in humans

Using current animal models, it is not possible to identify low-molecular-weight compounds (LMWCs) that are likely to be associated with anaphylaxis. It is generally accepted that the ultimate effector mechanism involves drug-induced IgE antibody. The objective of the present study was to determine if diclofenac, zomepirac and glafenine, which are associated with anaphylaxis in humans, have immunostimulating potential in the murine TNP-OVA (trinitrophenyl-ovalbumin) popliteal lymph node assay (PLNA), and more specifically to determine if the immunostimulation caused by these LMWCs results in IgE antibody production. These LMWCs were chosen because both zomepirac and glafenine were removed from the market due to high association with anaphylaxis, and diclofenac, which remains on the market, is frequently associated with anaphylaxis. In addition to conducting a TNP-OVA PLNA, the immunostimulating potential of these compounds was examined in the direct PLNA. When co-administered with TNP-OVA, all three LMWCs caused dose-dependent (0.25, 0.50, 1.00 and 1.25 mg) increases in popliteal lymph node (PLN) weight and cellularity that were observed beginning with the 0.25-mg dose. In addition, beginning with the 0.25-mg dose, all three compounds caused dose-dependent increases in TNP-OVA specific IgM and IgG(1) antibody-forming cells (AFCs). Diclofenac induced an isotype switch and caused a dose-dependent increase in the number of IgE AFCs with no detectable IgG(2a) AFCs and minimal high-dose-only IgG(2b) AFCs. Zomepirac induced IgE, IgG(2a) and IgG(2b) AFCs following the injection of 0.50 mg only, and glafenine induced IgE, IgG(2a) and IgG(2b) AFCs following the injection of 0.50-1.00 mg. In the direct PLNA, diclofenac caused dose-dependent increases in PLN weight and cellularity that were observed beginning with dose of 0.50 mg, whereas zomepirac failed to increase any PLN parameter and glafenine only increased the PLN weight. These results suggest that diclofenac, zomepirac and glafenine are immunostimulating LMWCs in the TNP-OVA PLNA with the potential to induce IgE antibody against a co-administered hapten-conjugate. Furthermore, these results suggest that the TNP-OVA PLNA offered significant advantages over the direct PLNA. Although it is not realistic to suggest that a single assay, based on a low number of test compounds, can identify all LMWCs with the potential to cause anaphylaxis in humans, these observations do demonstrate the potential utility of the PLNAs in examining LMWC-induced immunomodulation and support further development and investigation of the assays.     

Investigation of the dose-response relationship upon intraperitoneal administration of coumarin and 7-hydroxycoumarin on the carrageenan induced edema of the rat hind paw

The dose-response relationship upon intraperitoneal administration of coumarin (C) and 7-hydroxycoumarin (7HC) in rats was evaluated using the carrageenan induced edema of the hind paw. In a preliminary study C was indicated to reduce the 4-h edema at doses of 10 to 60 mg/kg i.p. Doses of 2.5 and 5 mg/kg were not effective. In the same study 7HC did not appear to be effective over a dose range of 0.01 to 20 mg/kg. A second study based on a completely randomized design indicated 7HC to be ineffective at doses of 2.5, 5, 10 and 20 mg/kg for edema measurements made at 3, 4, 5 and 6 h after the injury stimulus. C, however, was found to be effective at doses of 5, 10 and 20 mg/kg at 3, 4 and 5 h. C dose of 2.5 mg/kg was ineffective at all times and the 20 mg/kg dose appeared to be ineffective at the 6-h measurement. Resolution of the edema after 5 h did not appear to be enhanced by either C or 7HC administration as the rate of decay appeared to be parallel for controls and the treated animals. The edema model used here would possibly be very useful for further pharmacological investigation of the benzopyrones due to its simplicity, reproducibility and speed.     

Metabolic interactions between nortriptyline and thioridazine in rats

Plasma drug concentrations in rats were higher when nortriptyline was given in combination with thioridazine than when nortriptyline was given alone, regardless of whether the route of administration was po, ip, or iv. Nortriptyline concentrations in brain were also elevated, and the half-life of nortriptyline in brain was prolonged, after thioridazine administration. Although metabolites of thioridazine reacted in the assay for plasma nortriptyline, the elevated drug contents were found to be due to nortriptyline, not to thioridazine metabolites, by measurement of drug concentrations in brain (in which case thioridazine metabolites did not interfere) and by experiments with radioactive nortriptyline and thioridazine. Concentrations of radioactivity after [¹⁴C]nortriptyline administration were elevated in all tissues studies, especially in the liver, by concurrent administration of thioridazine. Hypothermia occurred in rats given large oral doses of thioridazine, but not nortriptyline; greater hypothermic effects were produced by the drug combination. Large ip doses of nortriptyline produced marked hypothermia and gave high plasma drug concentrations comparable to those found when oral nortriptyline was given in combination with thioridazine. The enhanced hypothermia found with the drug combination, therefore, seemed to be due to the combined pharmacologic effects of thioridazine and nortriptyline and probably relates to the enhanced lethality of the drug combination observed by others. The interaction at high doses is not limited to this tricyclic antidepressant-phenothiazine combination, for enhanced hypothermia was also observed when amitryptyline and perphenazine were coadministered. Although the results with large doses in laboratory animals may not have direct relevance to clinical doses of these psychotropic drugs, they do suggest a need for caution when tricyclic antidepressant drugs and phenothiazines are used in combination clinically.     

Synthesis and analgesic activity of some quinazoline analogs of anpirtoline

New condensed derivatives of anpirtoline, in which the pyridine ring is replaced with quinoline, quinazoline, 7-chloroquinoline, and 7-chloroquinazoline nuclei, have been synthesized. Their receptor binding profiles (5-HT1A, 5-HT1B) and analgesic activity (hot plate, acetic acid induced writhing) have been studied. The analgesic activity of compounds 4e-4g, and 4l are at least comparable to that of clinically used drugs flupirtine and tramadol under the same conditions.     

Metabolism of atenolol in man.

1. The disposition and metabolism of 1-(4-carbamoyl[14C]methylphenoxy)-3-isopropylaminopan-2-ol (atenolol, Tenormin) has been studied in man following oral and intravenous doses. 2. Approx. 50% of an oral dose was eliminated in urine; the major radiolabelled component was atenolol (approx. 90%). Faecal extracts also contained largely unchanged atenolol, with small amounts of more polar metabolites. Biliary excretion of atenolol and its metabolites is not a major route of elimination in man. Metabolism of the compound is not extensive and route-dependent modes of metabolism do not appear to complicate the position. 3. Atenolol appeared to be the only major radiolabelled component in blood. 4. Oral doses of atenolol are incompletely absorbed (range 46-62%), even when formulated as a solution. 5. 1-[4-(C-Carbamoylhydroxymethyl)phenoxy]-3-isopropylaminopropan-2-ol was a minor urinary metabolite, which has only one tenth the activity of the parent compound as a beta-adrenergic blocking agent in the rat. 6. Pharmacological activity in man appears to be due to atenolol alone.     

Comparative analgesic efficacy and tolerability of ketorolac tromethamine and glafenine in patients with post-operative pain

Summary

In a randomized, single-dose, double-blind, parallel comparative trial of analgesic efficacy, 96 adult patients received either 10?mg ketorolac tromethamine or 400?mg glafenine orally the morning after surgery if they requested pain relief medication. Each patient provided a baseline pain assessment and then received the assigned medication. Patients assessed pain intensity and pain relief and reported any adverse events in interviews held 30 minutes after drug administration and then hourly for 6 hours. The demographic characteristics, baseline pain intensity, and surgical categories of the 47 patients who received ketorolac tromethamine and the 49 who received glafenine were similar. Both drugs provided prompt, sustained pain relief throughout the 6-hour observation period, and there were no statistically significant differences between the two groups in any of the efficacy measures analyzed. The global assessment recorded by patients suggested a slight clinical advantage for ketorolac tromethamine (32.6% of ‘excellent’ responses) as compared to glafenine (12.5% ‘excellent’). The differences in overall response were statistically significant (p = 0.017). Fourteen (30%) patients who received ketorolac tromethamine and 17 (35%) who received glafenine reported adverse experiences that began or seemed to worsen after administration of the study drugs. The most prominent were drowsiness and sleeping, both of which are common in post-surgical patients.     

Importance of noradrenaline found in a functional pool in maintaining spontaneous locomotor activity in rats

1. Spontaneous locomotor activity (activity) in male Wistar rats was compared with the concentrations of brain noradrenaline (NA), dopamine (DA) and metaraminol.2. alpha-Methyl-m-tyrosine (alphaMMT) (400 mg/kg) reduced the concentrations of DA as well as NA but activity remained high in the presence of metaraminol formed from the alphaMMT. When tetrabenazine (TBZ) was given after alphaMMT pretreatment there was a fall in the levels of activity and in the concentrations of NA, DA and metaraminol.3. alpha-Methyl-p-tyrosine (alphaMT) produced a fall in activity which was correlated with falls in the concentrations of NA and DA. 5-Hydroxytryptamine (5-HT) did not appear to be affected.4. After depletion of NA and DA by alphaMT and TBZ, administration of L-dopa produced a return in activity which was significantly correlated with the concentration of NA but not DA. When alphaMMT was given to a similar group of pretreated animals there was no recovery of activity despite high concentrations of DA and metaraminol.5. The dopamine beta hydroxylase inhibitor, diethyldithiocarbamate (DDC), suppressed activity as well as the concentrations of NA and DA at high doses (750 mg/kg) but smaller doses (400 mg/kg) plus L-dopa gave high DA concentrations without activity.6. It is concluded that NA and not DA is associated with activity but that it is only part of the total NA which is in the biosynthetic storage granule affected by drugs like alphaMT and TBZ, which controls activity. Drugs that do not affect this pool may lower NA concentrations but not reduce activity.7. The replacement of NA by metaraminol in this functional pool does not restore activity.     

Synthesis and analgesic activities of some (4-substituted phenyl-1-piperazinyl)alkyl 2-aminobenzoates and 2-aminonicotinates.

A series of (4-substituted phenyl-1-piperazinyl)alkyl 2-aminobenzoates and 2-aminonicotinates has been prepared and screened for analgesic and antiinflammatory properties in mice and rats. The tabulated results reveal several 2-(4-substituted phenyl-1-piperazinyl)ethyl 2-(7- or 8-substituted 4-quinolinylamino)benzoates to be six to nine times more potent analgesics than the reference compounds (glafenine and aminopyrine) and to possess minor antinflammatory activity. Compound 45, 2-[4-[3-(trifluoromethyl)phenyl]-1-piperazinyl]ethyl 2-[[7-(trifluoromethyl)-4-quinolinyl]amino]benzoate (antrafenine), showed marked analgesic activity, long duration of action, and excellent tolerance in pharmacological and toxicological studies, as well as in clinical trials.     

(8-Chlor-3-ethyl-4-oxo-1,4-dihydro-7-chinolyloxy)-essigsäure,ein Reaktionsprodukt von Etacrynsäure mit Hydroxylamin

(8-Chloro-3-ethyl-1,4-dihydro-4-oxo-7-quinolyloxy)acetic Acid, a Product Obtained from Etacrynic Acid and Hydroxylamine A reaction for the determination of etacrynic acid (1) is proposed for the 2nd edition of the European Pharmacopoeia. It yields the fluorescent title substance 7B . The structure of 7B is determined by spectroscopic methods and by its conversion to the 4-chloroquinoline 8 .     

新抗疟药M6407的合成

A new drug, 4-(3′, 5′-bis-pyrrolidin-l-yl-methylene-4′-hydroxyphenyl)amino-7-chloroquinoline, was synthesized. After two years of pharmacological studies, it was found that the drug exhibited lower toxicity and bigher potency against Plasmodium berghei in mice when compared with chloroquine.     

Novel 4-aminoquinolines active against chloroquine-resistant and sensitive P. falciparum strains that also inhibit botulinum serotype A

We report on the initial result of the coupling of 4-amino-7-chloroquinoline with steroidal and adamantane constituents to provide small molecules with excellent in vitro antimalarial activities (IC90 (W2) down to 6.74 nM). The same entities also inhibit the botulinum neurotoxin serotype A light chain metalloprotease at low micromolar levels (7-31 microM). Interestingly, structural features imparting increased antimalarial activity also provide increased metalloprotease inhibition, thus allowing for simultaneous compound optimizations against distinct targets.     

New derivatives of benzopyran-4-one with analgesic, anti-inflammatory and antiplatelet clumping activity

A series of 35 new derivatives of 3-benzylidene benzopyran-4-one carboxylic acid was synthesized; their antiinflammatory and analgesic activities were investigated and compared with those of acetylsalicylic acid, phenylbutazone, ketoprofene and glafenine. Among these compounds, four were found to be more potent than the references compounds in tests of antiinflammatory and analgesic activity. Seven compounds were tested on rat blood platelet aggregation induced by ADP. From these tests, HD-039, HD-041, HD-044 and HD-049 appear to be promising antiinflammatory, analgesic and anticlotting agents, although this original series shows little structural analogy with classical non-steroidal antiinflammatory agents.     

Comparative analgesic efficacy and tolerability of ketorolac tromethamine and glafenine in patients with post-operative pain

In a randomized, single-dose, double-blind, parallel comparative trial of analgesic efficacy, 96 adult patients received either 10 mg ketorolac tromethamine or 400 mg glafenine orally the morning after surgery if they requested pain relief medication. Each patient provided a baseline pain assessment and then received the assigned medication. Patients assessed pain intensity and pain relief and reported any adverse events in interviews held 30 minutes after drug administration and then hourly for 6 hours. The demographic characteristics, baseline pain intensity, and surgical categories of the 47 patients who received ketorolac tromethamine and the 49 who received glafenine were similar. Both drugs provided prompt, sustained pain relief throughout the 6-hour observation period, and there were no statistically significant differences between the two groups in any of the efficacy measures analyzed. The global assessment recorded by patients suggested a slight clinical advantage for ketorolac tromethamine (32.6% of 'excellent' responses) as compared to glafenine (12.5% 'excellent'). The differences in overall response were statistically significant (p = 0.017). Fourteen (30%) patients who received ketorolac tromethamine and 17 (35%) who received glafenine reported adverse experiences that began or seemed to worsen after administration of the study drugs. The most prominent were drowsiness and sleeping, both of which are common in post-surgical patients.     

Synergistic interaction of a chloroquine metabolite with chloroquine against drug-resistant malaria parasites

We have previously shown that structural modification of chlorpromazine to introduce a basic side chain converts this chloroquine (CQ) resistance-reversing agent into a compound that has activity against Plasmodium falciparum in vitro. In an effort to further dissect the structural features that determine quinoline antimalarial activity and drug resistance-reversing activity, we have studied a series of aminoquinolines that are structurally related to CQ. We have analysed their haematin-binding activities, their antimalarial activities and their abilities to synergise the effect of CQ against drug-resistant P. falciparum. We found that a number of the aminoquinolines were able to interact with haematin but showed no or very weak antiparasitic activity. Interestingly, 4-amino-7-chloroquinoline, which is the CQ nucleus without the basic side chain, was able to act as a resistance-reversing agent. These studies point to structural features that may determine the resistance-modulating potential of weakly basic amphipaths. Interestingly, 4-amino-7-chloroquinoline is a metabolic breakdown product of CQ and may contribute to CQ activity against resistant parasites in vivo.     

抗疟药双咯喹及其类似物的合成

报道了双咯喹(4)和同类抗疟药阿莫地喹(1)、环喹(2)、阿莫吡喹(3)及衍生物5～14的合成,均用烷基胺与4-(4-羟基苯胺基)-7-氯喹啉进行Mannich反应而得。对鼠伯氏疟原虫的作用,以4及3最强;毒性均低于氯喹;与氯喹的交叉抗性指数分别为4及5。4治疗间日疟病人的疗效与氯喹相当。1,2,3,7,8还具有抗小鼠日本血吸虫的作用。     

Taloximine, a new respiratory stimulant with bronchodilator properties

1. A novel phthalazine analogue taloximine (1-hydroxyimino-4(2-dimethyl-aminoethoxy)-1,2-dihydrophthalazine monohydrochloride monohydrate) stimulated respiration in conscious rabbits at doses of 7 mg/kg and above.2. Taloximine antagonized the depressant action of morphine on respiration in rabbits at doses of 10 mg/kg. At high doses it resuscitated rabbits after they had been given lethal doses of sodium pentobarbitone.3. In in vitro preparations of the trachea or bronchus taloximine was about equiactive with aminophylline. In the Konzett-Rössler preparation the intravenous ED50 for taloximine was about 18% of that of aminophylline, whereas after oral administration the two drugs were equiactive.4. Taloximine, unlike aminophylline, did not protect guinea-pigs against anaphylactic microshock to egg albumen.5. Taloximine shortened the duration of the loss of righting reflex in mice due to hexobarbitone more effectively than bemegride, nikethamide or vanillic acid diethylamide.6. In the dose required to stimulate respiration taloximine had only slight cardiovascular effects. Up to four times this dose produced no evidence of general excitation of the central nervous system.     

Cytosolic myocardial calcium modulation by ATP-dependent potassium channel openers and NO donors

The goal of this study was to evaluate, in rat cardiomyocytes, the effects on cytosolic calcium of a pure K-adenosine triphosphate (ATP) channel opener, aprikalim, and those of nicorandil, a dual-acting agent that increases cyclic guanosine monophosphate (cGMP) levels and opens K-ATP channels. These effects were compared with those of a pure NO donor, 3-morpholino-sydnonimine (Sin-1). Ventricular myocytes were isolated from the hearts of adult rats. Changes in cytosolic calcium concentration ([Ca2+]i) were measured by using a Ca2+ indicator, indo-1/AM. Alterations in indo-1 fluorescence were recorded during regular electrical stimulation. After 10 min of pacing, end-diastolic [Ca2+]i was significantly increased as compared with control without significant changes in calcium transient. For doses of 10(-7) to 10(-4) M, aprikalim and nicorandil did not affect significantly the calcium transient. Sin-1 produced a significant decrease in calcium transient (by approximately 20%), which was already maximal at 10(-7) M. When given with the potassium channel antagonist glibenclamide (10(-5) M), nicorandil induced the same effects as those observed with Sin-1. We conclude that potassium channel openers aprikalim and nicorandil do not not decrease calcium transient. Thus the NO-donor properties of nicorandil are not apparent when given alone but appear when ATP-dependent potassium channels are blocked.     

Effects of TCDD on vitamin A status and liver microsomal enzyme activities in a TCDD-susceptible and a TCDD-resistant rat strain

To investigate the relationship between vitamin A status and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) lethality, the influence of TCDD on tissue and serum vitamin A levels was determined in the most TCDD-susceptible (Long-Evans) and the most TCDD-resistant (Han/Wistar) rat strains. The TCDD LD50 values of these two strains differ by a factor of more than 300. Groups of three rats per strain were used in a dose-response study (given single ip doses of 0, 4, 40, 400, 800 or 1600 micrograms TCDD/kg body weight and killed on day 11) and in a time-course experiment (given single ip doses of 0, 4 and, in the case of Han/Wistar rats only, 1600 micrograms TCDD/kg body weight, and killed on days 4, 11, 23, 50 and 76). The strains showed similar response over the 76-day study with respect to vitamin A levels in the liver, kidneys, testicles and serum after exposure to a sublethal dose of TCDD (4 micrograms/kg body weight). In contrast, TCDD doses lethal to the Long-Evans strain only (40-1600 micrograms/kg, day 11) markedly increased kidney and serum vitamin A levels in Han/Wistar rats, while they were practically without effect in Long-Evans rats. Hepatic cytochrome P-450 concentration, and the activities of 7-ethoxyresorufin O-deethylase, ethylmorphine N-demethylase, and uridine diphosphate glucuronosyltransferase (towards p-nitrophenol) were affected by the TCDD doses in much the same manner in both strains. These findings show that the correlations between TCDD lethality and changes in vitamin A status found among species of laboratory animals do not hold for Long-Evans and Han/Wistar strains of rat.     

Discriminating effects of a nucleotide-rich yeast extract,ProbioticumR, as an immunomodulator contrasted with actions in chronic immuno-inflammatory disease (adjuvant-induced arthritis) in rodents

Following previous observations that yeast ribonucleic acid (RNA) and nucleotide components exhibit immuno-stimulation in mice it was decided to determine the effects of a commercially-available yeast extract used for growth enhancement in domestic animals which is rich in RNA and nucleotides, Probioticum^R (Chemoforma AG, Augst, Switzerland) in standard immune models in mice and for effects on the progression of adjuvant-induced polyarthritis in rats. Probioticum (PB) (1 g kg⁻¹ day⁻¹) given p.o. for 6 weeks to mice, which were also immunized during the last 6 weeks with sheep red blood cells, produced activation of macrophages (detected by specific monoclonal antibodies) in cells of the bone marrow, increased antigen specific cells in the spleen, and a small reduction of thymus T-lymphocytes compared with control animals. High oral doses (up to 300–500 mg kg⁻¹ day⁻¹) of PB given alone or with indomethacin p.o. during the inductive or established phases of adjuvant-induced arthritis failed to affect the course of this disease. Thus, the immunological properties of PB appear principally to involve macrophage- and B-cell activation, with some possible minor alterations in T-cell activity. The immunological effects of PB do not appear to be important for the prevention of adjuvant arthritis nor do they influence the prostaglandin-dependent components of the disease influenced by indomethacin.     

Distribution of recombinant human erythropoietin following multiple intravenous administration and effects of age on the distribution in rats.

Multiple administration of 125I-recombinant human erythropoietin (125I-rh-EPO) did not cause an accumulation of intact EPO in tissue. The gel filtration profile of tissue from rats given multiple doses of 125I-rh-EPO was very similar to that for rats given single doses of 125I-rh-EPO, suggesting that intact rh-EPO (CAS 11096-26-7) does not accumulate. Pretreatment of rats with non-labeled rh-EPO did not cause a change in the ratio of metabolites to intact rh-EPO, as judged by the gel filtration data. This indicates that multiple administrations do not cause a change in metabolic pathways in the tissues. The tissue to plasma concentration ratio (t/p ratio) of rh-EPO in the target organ, i.e., the bone marrow and spleen, decreased with increase of age; the spleen showed a marked decrease in t/p ratio as compared to the bone marrow. On the other hand, t/p ratios in the kidney and liver did not change for periods of 4 to 31 weeks.