zaprinast and siguazodan

An increase of phosphodiesterase 4 (PDE4) has been described in blood mononuclear white cells of patients with atopic dermatitis (AD). This study was intended to search for the putative relationship between histamine and PDE4 in inflammatory cells.. The human monocyte cell line U-937 was used as a model of blood mononuclear leucocytes. PDE4 activity was determined as the increase of cAMP degradation, which is specifically inhibited by rolipram. Intracellular cAMP content was measured and correlated to PDE4 activity.. 1 microM histamine produced a 2- to 3-fold selective increase in PDE4 activity in U-937 cells after 4 hours of incubation. Enzyme activation was reversible, concentration- and time dependent. Cycloheximide (10 microM) prevented histamine-induced stimulation of PDE4. The H2-receptor antagonist, ranitidine, but not the H1-receptor antagonist, diphenhydramine, prevented histamine-induced activation of PDE4 in a concentration-dependent manner. Inhibition of cAMP degradation in the presence of histamine plus rolipram after 4 h of incubation further increased cAMP levels and PDE4 activity.. These results suggest that histamine-induced stimulation of PDE4 is mediated by the H2 receptor and related to intracellular levels of cAMP. AMPc enhancement of the cyclic nucleotide levels may trigger expression and synthesis of this enzyme.

Topics: 1-Methyl-3-isobutylxanthine; 3',5'-Cyclic-AMP Phosphodiesterases; Cyclic AMP; Cyclic Nucleotide Phosphodiesterases, Type 4; Diphenhydramine; Guanidines; Histamine; Humans; Kinetics; Purinones; Pyridazines; Ranitidine; Receptors, Histamine H2; Rolipram; U937 Cells

The present study was undertaken to investigate and compare responses to the cyclic nucleotide phosphodiesterase inhibitors siguazodan (type III, guanosine 3',5'-cyclic monophosphate (cGMP)-inhibited adenosine 3',5'-cyclic monophosphate (cAMP)), rolipram (type IV, cAMP-specific), and zaprinast (type V, cGMP-specific) in the feline pulmonary vascular bed. When tone in the pulmonary vascular bed was raised to a high steady level with a constant infusion of the thromboxane mimic U46619 (9,11-dideoxy-11, alpha9alpha-epoxymethano prostaglandin F(2alpha)), intralobar injections of the three phosphodiesterase inhibitors caused dose-related decreases in lobar arterial pressure. In terms of relative vasodilator activity, rolipram was more potent at higher doses than siguazodan, which was more potent than zaprinast. The duration of the pulmonary vasodilator response to zaprinast was shorter than for siguazodan or rolipram. Furthermore, siguazodan and rolipram, but not zaprinast, decreased systemic arterial pressure when injected into the perfused lobar artery in the range of doses studied. The present data demonstrate that the three phosphodiesterase inhibitors have potent, long-lasting vasodilator activity in the pulmonary vascular bed of the cat. These data suggest that there is rapid turnover of cAMP and cGMP in the pulmonary circulation and indicate that phosphodiesterase enzyme types III, IV, and V may play an important role in the regulation of vasomotor tone in the feline lung.

Topics: Animals; Blood Pressure; Cats; Dose-Response Relationship, Drug; Female; Guanidines; Male; Phosphodiesterase Inhibitors; Pulmonary Circulation; Purinones; Pyridazines; Rolipram

The possible interaction between spontaneously synthesized relaxant prostaglandins and the relaxation produced by three different isoenzyme-selective phosphodiesterase inhibitors was investigated in the isolated guinea-pig trachea in vitro. The relaxant action of siguazodan (phosphodiesterase III inhibitor), rolipram (phosphodiesterase IV inhibitor) and zaprinast (phosphodiesterase V inhibitor) was investigated in preparations with either spontaneously tone alone or in preparations with spontaneous tone and additionally stimulated with histamine (1 microM). In addition, relaxant effects were assessed in preparations without spontaneous tone (inhibited by indomethacin 2 microM) and precontracted with histamine (1 microM) or prostaglandin F2 alpha (10 microM), either alone or in the presence of a non-relaxant concentration (20 nM) of prostaglandin E2. All three phosphodiesterase inhibitors preferentially relaxed preparations with spontaneous tone and showed increased relaxant effects in preparations with spontaneous tone and additionally stimulated with histamine compared to preparations contracted by histamine alone. This enhanced relaxing effect observed in the presence of initial spontaneous tone was mimicked by exogenous application of prostaglandin E2 to indomethacin treated preparations either precontracted by histamine or prostaglandin F2 alpha. Furthermore, the study revealed marked differences in the relaxant profiles of siguazodan, rolipram and zaprinast, differences which most likely are related to the functional importance of the phosphodiesterase isoenzymes inhibited by these drugs. It is concluded that endogenously synthesized relaxant prostaglandins and exogenously applied prostaglandin E2 are capable of enhancing the relaxant action of the phosphodiesterase inhibitors siguazodan, rolipram and zaprinast and that cyclooxygenase inhibition is an important way to avoid this interaction in experimental studies of airway smooth muscle relaxants in isolated guinea-pig trachea in vitro.

Topics: Animals; Cyclooxygenase Inhibitors; Dinoprost; Dinoprostone; Drug Interactions; Female; Guanidines; Guinea Pigs; Histamine; In Vitro Techniques; Indomethacin; Male; Muscle Contraction; Muscle Relaxation; Muscle, Smooth; Oxytocics; Phosphodiesterase Inhibitors; Purinones; Pyridazines; Pyrrolidinones; Rolipram; Trachea

The ability of low concentrations of salbutamol to potentiate the relaxant effects of the phosphodiesterase (PDE) inhibitors, rolipram, Ro 20-1724 (PDE type IV inhibitor), siguazodan and milrinone (PDE type III inhibitor) was studied on guinea pig isolated trachea. These PDE inhibitors were strong relaxants of guinea pig trachealis under basal tone, but had only a weak activity on tissues precontracted with histamine (10(-5) M). In both cases, PDE type IV inhibitors showed a relaxant effect composed of two phases. The first phase represented 20 and 40% and the second, 90 and 140%, respectively, of relaxation of basal tone and histamine-induced tone. A second characteristic of PDE type IV inhibitors was the very fast and partially reversible relaxation observed at concentrations greater than 3 x 10(-8) M (for histamine-induced tone) at the first addition of inhibitor, followed by a residual relaxant activity. The latter relaxant effect was stable at concentrations of 3 x 10(-8)-10(-5) M and was equivalent to a 20% relaxation (for histamine-induced tone). In the presence of low concentrations (10(-9) and 10(-8) M) of salbutamol, there was a significant concentration-dependent potentiation of the effects of PDE inhibitors on trachea precontracted with histamine. Salbutamol, at a concentration of 10(-9) M, potentiated the effects of PDE inhibitors between 1.4- and 3.6-fold. In the presence of salbutamol 10(-8) M, the potentiation was more marked for siguazodan (37.9-fold), milrinone (11.0-fold) and Ro 20-1724 (14.5-fold) than for rolipram (4.3-fold). These results suggest that low concentrations of salbutamol can potentiate the relaxant effects of both PDE type III and PDE type IV inhibitors. Thus, PDE type IV inhibitors, which have antiinflammatory properties, could also provide adequate bronchodilation when used in combination with lower than usual doses of beta 2-agonists.

Topics: 4-(3-Butoxy-4-methoxybenzyl)-2-imidazolidinone; Adrenergic beta-Agonists; Albuterol; Animals; Drug Synergism; Guanidines; Guinea Pigs; Histamine; In Vitro Techniques; Male; Milrinone; Muscle Contraction; Muscle Relaxation; Muscle Tonus; Muscle, Smooth; Osmolar Concentration; Phosphodiesterase Inhibitors; Purinones; Pyridazines; Pyridones; Pyrrolidinones; Rolipram; Theophylline; Trachea

Antigen (ovalbumin)-induced contraction of guinea-pig isolated trachea, which largely resulted from the endogenous release of peptidoleukotrienes, was strongly inhibited by the non-selective phosphodiesterase (PDE) inhibitor theophylline and, more potently, by the selective PDE type IV inhibitors rolipram and Ro 20-1724. It was also strongly inhibited by the PDE type V inhibitor zaprinast, but much less so by the PDE type III inhibitor siguazodan and milrinone. Similar results were obtained in trachea minus epithelium. In contrast to their effects vs. allergic airway smooth muscle contraction, both milrinone and siguazodan potently relaxed leukotriene C4 (LTC4)-induced contraction in isolated trachea from non-sensitized animals. In this assay, rolipram, Ro 20-1724 and zaprinast were less active compared to their effects vs. ovalbumin-induced contraction, whereas theophylline had equivalent potency in the two tests. The relative potencies of rolipram and siguazodan in relaxation of trachea were similar when added prior to or after either LTC4 or histamine. These results suggest that the higher potency of selective PDE type IV & V inhibitors compared with PDE type III inhibitors vs. ovalbumin-induced contraction is due to their greater inhibition of anaphylactic mediator release. The converse is true if we consider their bronchodilator actions, although the superior efficacy of selective PDE type III inhibitors over PDE type IV inhibitors may vary in sensitized vs. non-sensitized animals. The present results are in agreement with a previous study showing that low concentrations of a beta 2-agonist increased the relaxant effect of selective PDE type IV inhibitors in guinea-pig trachea. The present data indicate that prophylactic use of selective PDE type IV inhibitors combined with therapeutic use of low dose inhaled beta-agonist might represent an alternative to the use of antiallergic or steroid therapy in asthma.

Topics: 4-(3-Butoxy-4-methoxybenzyl)-2-imidazolidinone; Animals; Bronchoconstrictor Agents; Bronchodilator Agents; Guanidines; Guinea Pigs; Histamine Antagonists; In Vitro Techniques; Leukotriene C4; Male; Milrinone; Muscle Tonus; Ovalbumin; Phosphodiesterase Inhibitors; Purinones; Pyridazines; Pyridones; Pyrrolidinones; Rolipram; Theophylline; Trachea

Nonadrenergic, noncholinergic relaxations were elicited by field stimulation (1-16 Hz, 1 msec, 8 V for 15 sec) of guinea pig trachea desensitized with capsaicin (3 microM), pretreated with atropine (1 microM), propranolol (1 microM), indomethacin (3 microM) and treated with alpha-chymotrypsin (2 U/ml) and contracted with 3 microM histamine. The effect of the phosphodiesterase (PDE) isozyme selective inhibitors siguazodan (PDE III-selective), rolipram (PDE IV-selective), denbufylline (PDE IV-selective) and zaprinast (PDE V-selective) was examined on the relaxant responses to field stimulation and on relaxations elicited by the nitric oxide donor 3-morpholinosydnonimine-N-ethylcarbamide (SIN-1). The response to field stimulation in the presence of alpha-chymotrypsin (the putative nitric oxide component), at all the frequencies tested, was potentiated significantly by the PDE IV inhibitors rolipram (1 and 10 microM) and denbufylline (3 and 10 microM) as were responses to SIN-1. The PDE V inhibitor zaprinast (30 microM) potentiated relaxations elicited by field stimulation at 8 and 16 Hz and also potentiated responses to SIN-1. The PDE III inhibitor siguazodan (1 microM), however, was without effect on relaxant responses to field stimulation or to SIN-1. These results suggest that the nitric oxide component of the nonadrenergic, noncholinergic relaxant response is mediated primarily via cyclic AMP whose action is inactivated by a PDE IV isozyme and also by cyclic GMP which is inactivated by a PDE V isozyme.

Topics: 3',5'-Cyclic-AMP Phosphodiesterases; Animals; Atropine; Cyclic AMP; Cyclic GMP; Cyclic Nucleotide Phosphodiesterases, Type 3; Cyclic Nucleotide Phosphodiesterases, Type 4; Electric Stimulation; Guanidines; Guinea Pigs; Indomethacin; Isoenzymes; Muscle Relaxation; Nitric Oxide; Phosphodiesterase Inhibitors; Phosphoric Diester Hydrolases; Propranolol; Purinones; Pyridazines; Pyrrolidinones; Receptors, Adrenergic; Receptors, Cholinergic; Rolipram; Trachea; Vasoactive Intestinal Peptide; Xanthines

1. We examined various type-selective phosphodiesterase (PDE) inhibitors on glucose-induced insulin secretion from rat isolated islets, on islet PDE activity and on islet cyclic AMP accumulation in order to assess the relationship between type-selective PDE inhibition and modification of insulin release. 2. The non-selective PDE inhibitor, 3-isobutyl-1-methylxanthine (IBMX, 10(-5)-10(-3) M), as well as the type III selective PDE inhibitors SK&F 94836 (10(-5)-10(-3) M), Org 9935 (10(-7)-10(-4) M), SK&F 94120 (10(-5)-10(-4) M) and ICI 118233 (10(-6)-10(-4) M) each caused concentration-dependent augmentation (up to 40% increase) of insulin release in the presence of a stimulatory glucose concentration (10 mM), but not in the presence of 3 mM glucose. 3. Neither the type IV PDE inhibitor rolipram (10(-4) M) nor the type I and type V PDE inhibitor, zaprinast (10(-4)-10(-3) M) modified glucose-induced insulin release when incubated with islets, although a higher concentration of rolipram (10(-3) M) inhibited secretion by 55%. However, when islets were preincubated with these drugs followed by incubation in their continued presence, zaprinast (10(-6)-10(-4) M) produced a concentration-dependent inhibition (up to 45% at 10(-4) M). Under these conditions, rolipram inhibited insulin secretion at a lower concentration (10(-4) M) than when simply incubated with islets. 4. A combination of SK&F 94836 (10(-5) M) and forskolin (5 x 10(-8) M) significantly augmented glucose-induced insulin secretion (30% increase), although neither drug alone, in these concentrations, produced any significant effect. 5. Islet cyclic AMP levels, which were not modified by forskolin (10-6 M), SK&F 94836 (10-4 M) or Org 9935 (10-5 M) were significantly elevated (approximately 3.7 fold increase) by forskolin inc ombination with either SK&F 94836 or Org 9935.6 Homogenates of rat islets showed a low Km (1.7 microM) and high Km (13 microM) cyclic AMP PDE in the supernatant fractions (from 48,000 g centrifugation), whereas the particulate fraction showed only a low Km (1.4 microM) cyclic AMP PDE activity.7. The PDE activity of both supernatant and pellet fractions were consistently inhibited by SK&F94836 or Org 9935, the concentrations required to reduce particulate PDE activity by 50% being 5.5 and 0.05 microM respectively.8 Rolipram (10-5 10-4 M) did not consistently inhibit PDE activity in homogenates of rat islets and zaprinast (10-4 M) consistently inhibited activity by 30% in the supernata

Topics: 1-Methyl-3-isobutylxanthine; 3',5'-Cyclic-AMP Phosphodiesterases; Analysis of Variance; Animals; Colforsin; Cyclic AMP; Dose-Response Relationship, Drug; Drug Interactions; Glucose; Guanidines; Insulin; Insulin Secretion; Islets of Langerhans; Isoenzymes; Male; Phosphodiesterase Inhibitors; Phosphoric Diester Hydrolases; Purinones; Pyrazines; Pyridazines; Pyrrolidinones; Rats; Rats, Sprague-Dawley; Rolipram; Thiophenes

We studied the effect of inhibiting phosphodiesterase (PDE) isozyme types III, IV and V on the cholinergic and noncholinergic (tachykinergic) contractile responses to electrical field stimulation (EFS) in the guinea pig isolated bronchus. SKF 94836, a PDE III inhibitor, had a slight (approximately 30%) but significant inhibitory effect on the noncholinergic contractions. Rolipram, an inhibitor of PDE IV isozymes, dramatically inhibited the noncholinergic contractions by nearly 70%. The EC50 for rolipram was approximately 20 nM. Rolipram (1 microM) had no effect on contractions elicited by either capsaicin or neurokinin A. EFS, but not direct vagus nerve stimulation, elicits small nonadrenergic, noncholinergic relaxations of the bronchus that were potentiated by rolipram. Rolipram had the same inhibitory effect on EFS- and vagus nerve stimulation-induced noncholinergic contractions. The effect of rolipram was mimicked by another PDE IV inhibitor, RO-201724. Inhibition of PDE V with zaprinast (3 microM) had no effect on the tachykinergic contractile response. None of the PDE inhibitors affected the EFS-induced cholinergic contractions. The data suggest that the contraction due to stimulation of tachykinergic fibers is significantly reduced by selective inhibition of PDE IV and, to a lesser extent, PDE III isozymes. This is unlikely to be due to functional antagonism at the level of the smooth muscle. It is also unlikely to be due to potentiation of the nonadrenergic relaxant response to nerve stimulation. Rather, the data are in agreement with the hypothesis that selective inhibition of PDE isozymes leads to inhibition of electrically evoked tachykinin release from capsaicin-sensitive fibers in the guinea pig bronchus.

Topics: 4-(3-Butoxy-4-methoxybenzyl)-2-imidazolidinone; Animals; Bronchi; Bronchoconstriction; Electric Stimulation; Guanidines; Guinea Pigs; Isoenzymes; Male; Phosphodiesterase Inhibitors; Purinones; Pyridazines; Pyrrolidinones; Rolipram; Vagus Nerve

Theophylline has been reported to inhibit excitatory noncholinergic but not cholinergic-neurotransmission in guinea-pig bronchi. As theophylline might exert this effect through an inhibition of phosphodiesterases (PDE), and since many types of PDE have now been described, the aim of this study was to investigate the effects of three specific inhibitors of PDE on the electrical field stimulation (EFS) of the guinea-pig isolated main bronchus in vitro. The drugs used were siguazodan, rolipram and zaprinast, which specifically inhibit PDE types, III, IV and V, respectively. Guinea-pig bronchi were stimulated transmurally with biphasic pulses (16 Hz, 1 ms, 320 mA for 10 s) in the presence of indomethacin 10(-6) M and propranolol 10(-6) M. Two successive contractile responses were observed: a rapid cholinergic contraction, followed by a long-lasting contraction due to a local release of neuropeptides from C-fibre endings. Rolipram (10(-9) to 10(-6) M) but not siguazodan or zaprinast, inhibited the peptidergic contraction in a concentration-dependent manner. Conversely, the cholinergic response was unaffected. Contractile responses induced by exogenous acetylcholine (10(-8) to 10(-3) M) or [Nle10]NKA(4-10) (10(-10) to 10(-6) M) were also unaffected by rolipram, siguazodan and zaprinast (10(-7) M). These results demonstrate that concentrations of rolipram, similar to those which inhibit PDE, reduce the release of sensory neuropeptides from C-fibre endings, and suggest that the cyclic adenosine monophosphate (AMP) PDE type IV is specifically involved in this effect, as in other anti-inflammatory effects.

Topics: Animals; Bronchi; Electric Stimulation; Female; Guanidines; Guinea Pigs; In Vitro Techniques; Male; Nerve Fibers; Phosphodiesterase Inhibitors; Purinones; Pyridazines; Pyrrolidinones; Rolipram; Synaptic Transmission