Efficacy and tolerability of hyaluronic acid, tea tree oil and methyl-sulfonyl-methane in a new gel medical device for treatment of haemorrhoids in a double-blind, placebo-controlled clinical trial.

Topical formulations are widely used in anti-haemorrhoidal treatment, but often lacking controlled clinical trials. Here, we report the results from a controlled clinical trial performed with a new gel medical device (Proctoial) containing hyaluronic acid with tea tree oil and methyl-sulfonyl-methane as major components. The total number of 36 haemorrhoidal patients (grade 1-3) was enrolled in a double-blind, placebo-controlled clinical trial and divided into 2 equal parallel groups. The anal pain, pain during defecation, visible bleeding, pruritus and irritation/inflammation were recorded before and after 14-day treatment using a visual analogue scale both by the investigators and by the patients. Safety and tolerability of the treatments were also recorded. The new gel medical device statistically significantly reduced all the symptoms after the treatment compared to placebo. The results indicated also a very good tolerability and safety of the treatments.

Protective activity of ketoprofen lysine salt against the pulmonary effects induced by bradykinin in guinea-pigs.

We investigated the capacity of ketoprofen lysine salt (KLS) to counteract the pulmonary effects of some mediators of airway inflammation. The protective effect of KLS and its R-isomer against bradykinin (BK) induced plasma extravasation in the airways and bronchoconstriction was evaluated in anaesthetized guinea-pigs, in parallel with the capacity of KLS to inhibit the production of thromboxane A2 (TXA2). Moreover, we studied the ability of KLS to modulate leukotriene C4 (LTC4) and acetylcholine (ACH) induced bronchoconstriction and the associated production of TXA2. Nimesulide (NIM) was used as the reference compound. KLS dose-dependently inhibited the bronchoconstriction and the associated production of TXA2 induced by BK, with closely related ID50 values of 31.2 and 34.0 micrograms/kg i.v., respectively. The protection was evident 10 min after KLS administration and, at 100 micrograms/kg i.v., lasted up to 2h, Moreover, KLS dose-dependently inhibited the increase in capillary permeability induced by BK, with a potency (ID50 23.4 micrograms/kg i.v.) slightly higher than that shown against the bronchoconstriction. KLS also prevented the bronchoconstriction and TXA2 production triggered by LTC4, but not ACH induced bronchoconstriction. In all the models studied, KLS was about 10 times more potent than NIM. These data demonstrate the capacity of KLS to counteract the bronchoconstriction induced by BK and LTC4 and to a large extent the airway inflammation induced by BK. Blockade of prostanoid production is likely to account for this protective effect, since the R-isomer of KLS was devoid of significant activity.

Effectiveness of carbocysteine lysine salt monohydrate on models of airway inflammation and hyperresponsiveness.

We investigated the possible effects of the mucoactive drug Carbocysteine lysine salt monohydrate (CLS.H2O) on experimentally-induced airway inflammation and hyperresponsiveness. CLS.H2O given by the oral route (300 mg kg(-1)) significantly reduced neutrophil infiltration into the airway lumen induced by intratracheal injection of IL-1 beta in rats. In addition, CLS.H2O inhibited dose-dependently (100-300 mg kg(-1) p.o.) the formation of pleural exudate and leukocyte recruitment induced by intrapleural injection of carrageenan in rats. Because of the close interaction between the inflammatory process and the development of airway hyperresponsiveness we also tested CLS.H2O on cigarette-smoke-induced inflammation and hyperreactivity in anaesthetized guinea-pigs. The drug, given either by oral (300 mg kg(-1)) or aerosol route (30-100 mg ml(-1)), was able to reduce the increase in airway responsiveness induced by smoke and the associated cell recruitment detected in the bronchoalveolar lavage (BAL) fluids. These results suggest that CLS.H2O can exert an anti-inflammatory action in addition to its mucoregulatory activity. The anti-inflammatory and anti-hyperreactivity effect of the drug within the airways may be of advantage in the treatment of inflammatory lung diseases where mucus secretion together with airway inflammation and hyperreactivity contribute to airway obstruction.

Effects of a new foam formulation of ketoprofen lysine salt in experimental models of inflammation and hyperalgesia.

The anti-inflammatory and analgesic profile of a new topical foam formulation of ketoprofen lysine salt (CAS 57469-78-0, Artrosilene Schiuma, KLS-foam) was characterized in comparison with marketed gel formulations containing KLS (KLS-gel) or diclofenac diethylammonium salt (DCF-gel). KLS-foam dose-dependently inhibited oedema formation and hyperalgesia induced by subplantar injection of carrageenan or substance P, being more potent than KLS-gel. At equieffective anti-inflammatory doses, KLS-foam provided a more pronounced analgesic effect than DCF-gel. KLS-foam also markedly inhibited exudate formation and prostaglandin production induced by subcutaneous implantation of carrageenan soaked sponges. In carrageenan induced paw inflammation, KLS-foam provided the same anti-inflammatory effect as orally administered KLS, but induced significantly less gastric damages. Oral administration of KLS resulted in sustained systemic absorption of ketoprofen, whereas after topical application of KLS-foam no appreciable ketoprofen plasma levels were detected. These data support the anti-inflammatory and particularly the analgesic effectiveness of the new foam formulation of KLS, a finding that, together with the high gastric tolerability, further emphasizes the usefulness of KLS-foam in the treatment of localized flogistic diseases and associated pain.

Effect of S-carboxymethylcysteine lysine salt on mucociliary clearance in rabbits with secretory cell metaplasia.

A single intratracheal instillation of porcine pancreatic elastase (PPE, 100 U/Kg) induces in rabbits bronchial secretory cell metaplasia as well as emphysematous changes. The mucus hypersecretion and the marked reduction of ciliated cells matched by a high percentage of atypical cilia are responsible for the delayed mucociliary clearance in this model. S-Carboxymethylcysteine lysine salt (SCMC-LYS, 0.35 g/Kg b.w.), given per os daily for 10 days starting 2 days before elastase administration, significantly ameliorated the mucociliary clearance. The pharmacological treatment did not modify the degree of secretory cell metaplasia and the percentage of atypical cilia, or prevent the alveolar wall destruction. At TEM examination, the morphological aspects of secretion occurring in bronchial tree of PPE-treated animals were rarely visible in the PPE + SCMC-LYS treated group. The beneficial effect of SCMC-LYS on mucociliary clearance may be ascribed to an antisecretagogue effect of this drug through elastase inhibition and to a reduction of mucus viscosity.

Stimulation of Cl- secretion by the mucoactive drug S-carboxymethylcysteine-lysine salt in the isolated rabbit trachea.

Ion transport by the airway epithelium contributes to the regulation of the quantity and composition of respiratory tract fluid, thereby affecting mucociliary clearance. We have investigated the effect of the mucoactive drug S-carboxymethylcysteine-lysine salt (S-CMC-Lys) on the transepithelial bioelectric properties of isolated rabbit trachea. Transepithelial potential difference (Vms), short-circuit current (Isc) and resistance (R) were measured in the isolated rabbit trachea mounted between flux half-chambers, in the presence and in the absence of S-CMC-Lys (100 microM), added to the mucosal or submucosal chamber. In some experiments, tissues were also exposed to ion channel-inhibitors, in order to evaluate the contribution of Na+ and Cl- active transport to Isc. The excised rabbit trachea expressed transepithelial bioelectric properties based on an active ion transport supported by the Na(+)-K(+)-adenosine triphosphatase (ATPase) activity, since ouabain (500 microM) completely abolished the transepithelial potential difference. In control preparations, Vms and Isc declined significantly during 300 min recording, whereas R remained constant. The Isc decline was essentially attributable to a decrease in Cl- transport. Bumetanide (100 microM) almost completely abolished the Isc fraction related to Cl- transport. Treatment of the tissues with S-CMC-Lys reduced the progressive fall in Isc, with the most clear-cut and significant effect observed for the mucosal treatment. In parallel, S-CMC-Lys significantly lowered R, without affecting Vms. Either mucosal or submucosal exposure to S-CMC-Lys significantly increased Cl- secretion to normal values, whilst Na+ absorption was not modified.(ABSTRACT TRUNCATED AT 250 WORDS)

Levodropropizine reduces capsaicin- and substance P-induced plasma extravasation in the rat trachea.

We investigated the effect of the non-opioid, peripherally acting antitussive agent levodropropizine to reduce neurogenic plasma extravasation in the rat trachea. Levodropropizine (10, 50 and 200 mg/kg) reduced in a dose-dependent manner the extravasation of Evans blue dye evoked by capsaicin. Levodropropizine inhibited also substance P-evoked extravasation, whereas it did not affect the extravasation evoked by platelet activating factor. Levodropropizine (10 and 100 microM) did not affect the contraction produced by [Sar9,Met(O2)11]substance P, a selective agonist for tachykinin NK1 receptors, in the rat urinary bladder in vitro. These data indicate that levodropropizine inhibits capsaicin-induced plasma extravasation: (a) acting at a postjunctional level; (b) exhibiting neuropeptide selectivity and; (c) via a mechanism independent of tachykinin NK1 receptor blockade. Irrespective of the mechanism, this novel antiinflammatory action of levodropropizine underlines its potential role in inflammatory airway diseases such as bronchial asthma.

Virus-induced airway inflammation and hyperresponsiveness in the guinea-pig is inhibited by levodropropizine.

Intratracheal Parainfluenza type 3 (PI-3) virus inoculation of guinea pigs leads to a non-specific airway hyperresponsiveness in vivo and in vitro which coincides with a significant increase in the number of inflammatory cells in the broncho-alveolar lavage fluid (90% increase, 4 days after inoculation). The activity of the bronchoalveolar cells, as measured by the chemiluminescence production of infected animals is significantly diminished (34.2%, 4 days after inoculation) after renewed stimulation with PI-3 virus in vitro as compared to the chemiluminescence production by bronchoalveolar cells obtained from control guinea pigs. Pretreatment of the guinea-pigs with the antitussive agent levodropropizine, administered intra-peritoneally twice a day for five successive days at a dose of 10 mg/kg, prevents the virus-induced airway hyperresponsiveness in vivo and in vitro, and inhibits the influx of broncho-alveolar cells. Levodropropizine at a dose of 1 mg/kg did not modulate these responses. Further, the decrease in chemiluminescence production of broncho-alveolar cells obtained from virus-infected animals after PI-3 virus stimulation in vitro was inhibited by levodropropizine (10 mg/kg). These data demonstrate the ability of levodropropizine to counteract the hyperresponsiveness phenomenon and the associated inflammatory event induced by PI-3 virus, an effect which may be due to its capacity to act on the peptidergic system or may be due to the anti-allergic/bronchoconstrictor property of this compound.

Effectiveness of levodropropizine against cigarette smoke-induced airway hyperreactivity: possible mechanism.

We verified the possible effect of the new antitussive drug levodropropizine on airway hyperreactivity and lung inflammation induced by cigarette smoke exposure in anaesthetized guinea-pigs. Levodropropizine, administered by aerosol at 25 mg/ml for 30 s completely prevented smoke induced airway hyperreactivity. The protective effect was early in onset (3 min) and lasted up to 30 min. The same dose of codeine, administered in the form of an aerosol, decreased the increase in airway responsiveness induced by smoke inhalation slightly but not significantly. In parallel with the functional results, levodropropizine also inhibited the recruitment of inflammatory cells triggered by smoke exposure within the airway lumen. When levodropropizine was administered i.v. to anaesthetized guinea-pigs, it reduced the bronchocontractile effect of capsaicin dose-dependently, whereas it was without effect against substance P-induced bronchoconstriction. These data demonstrate the ability of levodropropizine to counteract the hyperreactive phenomenon and the associated inflammatory event induced by cigarette smoke exposure, an effect which might depend on its capacity to modulate the activation of the peptidergic system.

Changes in airway reactivity to exogenous and endogenous acetylcholine and substance P after anaphylactic bronchoconstriction in anaesthetized guinea-pigs.

1. In anaesthetized, actively sensitized guinea-pigs, the anaphylactic shock induced by antigen aerosol challenge (5 s; 50 mg ml-1) was followed by increase in airway reactivity to both acetylcholine and substance P. In particular dose-response curves to acetylcholine (3-1000 micrograms kg-1 i.v.) and to substance P (5-80 micrograms kg-1 i.v.) obtained in antigen exposed animals were significantly shifted to the left of those performed in control guinea-pigs (exposed to saline aerosol). 2. The hyperreactive phenomenon after antigen aerosol was also evident when capsaicin-induced bronchoconstriction (1-4 micrograms kg-1 i.v.) was tested; the degree of hyperresponsiveness was similar to that observed with acetylcholine and substance P as agonists. 3. The frequency-response curves to vagal stimulation, either cholinergic or NANC in nature, were not significantly modified in guinea-pigs challenged with the antigen in respect to those aerosolized with saline. 4. The data obtained in the present study indicate that the airway hyperresponsiveness present in the animal model used is non-specific, involving both cholinergic and peptidergic effects. On the other hand, the lack of potentiation of the bronchoconstriction response to electrical stimulation might suggest that the establishment of a clear hyperreactive phenomenon is under the control of different mechanisms unrelated to increased bronchial reactivity.

Peripheral site of action of levodropropizine in experimentally-induced cough: role of sensory neuropeptides.

The mechanism of action of levodropropizine has been investigated in different models of experimentally-induced cough in guinea-pigs. In particular it has been demonstrated that the antitussive drug has a peripheral site of action by injecting the drug intracerebroventricularly (i.c.v.). In these experiments levodropropizine (40 micrograms/50 microliters i.c.v.) did not prevent electrically-induced cough. On the other hand, codeine (5 micrograms/50 microliters i.c.v.) markedly prevented coughing. A difference in the potency ratio of levodropropizine and codeine has been demonstrated in capsaicin-induced cough; after oral administration, codeine was about two to three times more potent than levodropropizine. However, after aerosol administration the two compounds were equipotent. These data might suggest a peripheral site of action for levodropropizine which is related to sensory neuropeptides. Further support for the role of sensory neuropeptides in the mechanism of action of levodropropizine comes from the results obtained in capsaicin-desensitized animals. In this experimental model levodropropizine failed to prevent the vagally elicited cough in neuropeptide-depleted animals, whereas codeine did not differentiate between control and capsaicin-treated animals. In conclusion, our results support the suggestion that levodropropizine has a peripheral site of action. In addition, the interference with the sensory neuropeptide system may explain, at least in part, its activity in experimentally-induced cough.

Nedocromil sodium prevents airway hyperreactivity induced by cigarette smoke in anaesthetized guinea-pigs.

Increased airway reactivity and influx of inflammatory cells into the airways have been demonstrated both in smokers and after smoke exposure in animal studies. We investigated the ability of nedocromil sodium and hydrocortisone to protect from the pathological alterations induced by direct cigarette smoke exposure in anaesthetized guinea-pigs. Active inhalation of cigarette smoke (15 s/min for 10 min) induced airway hyperreactivity, as shown by the enhanced bronchoconstrictor effect of histamine and was associated with an increase in total cells, macrophages and eosinophils in the BAL fluid. Nedocromil sodium given by aerosol (3 and 10 mg/ml for 30 s) completely prevented the ability of cigarette smoke to potentiate histamine induced bronchoconstriction. In parallel, nedocromil sodium inhibited the development of the inflammatory reaction triggered by smoke exposure. Hydrocortisone pretreatment (50 mg/kg s.c. twice) did not abolish the smoke induced airway hyperreactivity, nor did it inhibit the recruitment of proinflammatory cells within the airway lumen. Sensory neuropeptides have been demonstrated to be involved in the development of smoke induced airway hyperreactivity. The efficacy of nedocromil sodium in this model might depend on its ability to modulate the activation of the peptidergic system.

Interleukin-1 beta: a possible mediator of lung inflammation and airway hyperreactivity.

Interleukin-1 (IL-1), a peptide released from monocytes/macrophages, plays an important role in the inflammatory and immune responses. Airway hyperreactivity and the underlying airway inflammation are common features in asthma pathology. We investigated and characterized the inflammatory alterations induced within the guinea-pig respiratory system by IL-1 beta. Injection of IL-1 beta (1-6 micrograms/animal) into the pleural space resulted in a dose-dependent inflammatory response, as shown by the formation of pleural exudate and leucocyte recruitment. A threshold dose of IL-1 beta (1 micrograms/animal) markedly potentiated the inflammatory reaction triggered by the classical proinflammatory agent croton oil, underlining the amplifying role of IL-1 beta in the inflammatory events. The inflammatory process induced by intrapleural injection of IL-1 beta (6 micrograms/animal) was associated with the development of a hyperreactive phenomenon which involved both the peripheral and large airways. In fact, increased contractile activity of histamine was evident in the tracheas and parenchymal strips isolated from guinea-pigs exposed to IL-1 beta. These results provide evidence for a possible role of IL-1 beta in the genesis of airway inflammation and bronchial hyperreactivity.

Adrenaline-induced tracheal relaxation ex vivo is depressed following aerosol antigen challenge of sensitized guinea-pigs.

We investigated the changes in beta-adrenoceptor responses induced in guinea-pig tracheal and cardiac tissues by the anaphylactic reaction. Antigen aerosol challenge in sensitized guinea-pigs resulted in a marked reduction in adrenaline relaxation in isolated trachea ex vivo. The isoprenaline effect was also slightly decreased by antigen exposure, suggesting a possible impairment of tracheal beta-adrenoceptor function. On the other hand, the chronotropic and inotropic activity of adrenaline in isolated atria was not modified by the anaphylactic shock, suggesting a specific involvement of lung beta-adrenoceptors in the allergic reaction.

Differences in activity between noradrenaline and other alpha-agonists in rat vas deferens.

1 The stimulating activity of methoxamine on rat vas deferens differed from that of noradrenaline since it induced a strong rhythmic activity which was not removed by the wash-out of the drug. 2 Clonidine showed a dose-response curve with a pD2 of 5.05 +/- 0.14 and an intrinsic activity value of 0.6 +/- 0.1%; B-HT 920, a specific alpha 2-agonist, elicited a very low stimulating effect (pD2 = 2.87 +/- 0.04; i.a. = 0.08 +/- 0.001). 3 In a calcium-free medium the maximum responses to synthetic alpha-adrenoceptor agonists were reduced by 98 +/- 0.8% compared with the control value. The residual response to noradrenaline, however, was significantly higher (15 +/- 0.9% of the control value). 4 At high concentration of the Ca-channel antagonists, nicardipine and verapamil, only noradrenaline showed a residual response that was resistant to the calcium channel blockers. This residual response was completely inhibited by chloroethylclonidine (10(-5) M). 5 It is proposed that the stimulating activity of the physiological adrenergic agonist, noradrenaline, is more complex when compared to that of synthetic agonists and it might result from an interaction with different alpha-adrenoceptors. 6 Both salbutamol and forskolin were able to abolish the rhythmic activity of methoxamine, suggesting a regulatory role of cAMP on membrane stability.

Comparative study of postsynaptic alpha-adrenoceptors in aorta obtained from human and other mammalian species.

The alpha-adrenoceptor populations in aortic strips from humans, rats, guinea-pigs and rabbits were investigated in vitro, using specific agonists and antagonists. In rabbit and human preparations the activities of the two agonists tested, noradrenaline and methoxamine, were competitively antagonized by prazosin, whereas in the other animal species prazosin showed a competitive antagonism for methoxamine induced contraction, but an uncompetitive behaviour against noradrenaline. The alpha 2-selective agonists B-HT 920 and detomidine did not elicit any effect on aortic strips up to 10(-3) M. On the basis of these results, alpha 1-receptors seem to represent an homogeneous population in human and rabbit aortae, but not in rat and guinea-pig tissues. On the other hand, the contractile response of noradrenaline and methoxamine on aortic strips from the four animal species examined cannot be ascribed to the activation of postsynaptic alpha 2-receptors. Moreover we present preliminary evidence that rat and guinea-pig aortae do not contain a clear subdivision in alpha 1a- and alpha 1b-receptors.

Ganglion-blocking agents enhance neurally mediated bronchoconstriction in the guinea-pig: possible role of sensory neuropeptides.

The effect of ganglion blockade by hexamethonium bromide (0.1-100 mumol.kg-1) and pentolinium tartrate (0.01-3 mumol.kg-1) on the bronchoconstriction induced by vagal nerve stimulation (15 Hz, 0.2 ms, 3 s, 7-20 V) was evaluated in the anaesthetized guinea-pig. Both ganglion-blocking agents potentiated this response dose dependently. When the neural bronchoconstriction was suppressed by atropine, hexamethonium restored this response dose dependently. Hexamethonium produced inhibitory effects on vagally induced bronchoconstriction in capsaicin-desensitized and in propranolol- or reserpine-pretreated guinea-pigs. Propranolol (0.03-3 mumol.kg-1) produced a marked dose-dependent increase of neural bronchoconstriction (which was markedly reduced, about 10 times) in capsaicin-desensitized animals. Our results show that ganglion-blocking agents potentiate neural bronchoconstriction in the guinea-pig and that sensory neuropeptides may have a role in this effect. Moreover, beta-adrenergic modulation of the release of neuropeptides from vagal sensory fibers is suggested.

Mechanism of the potentiation of neurally-induced bronchoconstriction by gallamine in the guinea-pig.

1. Electrical stimulation of the cervical vagi (15 Hz, 0.2 ms, 3 s, 7-15 V) produced a slight bronchoconstriction in the anaesthetized guinea-pig. This effect was fully abolished by atropine, while gallamine (0.1-10 mumol kg-1) produced a dose-dependent increase up to ten fold. 2. Gallamine-induced potentiation of neurally-mediated bronchoconstriction was not inhibited by depletion of sensory neuropeptides with capsaicin or by pretreatment with pyrilamine. In propranolol-pretreated guinea-pigs the potentiation induced by gallamine 3 and 10 mumol kg-1 was inhibited by 40 and 46%, respectively. 3. Physostigmine (0.5 mg kg-1) produced a very slight and slowly developing bronchoconstriction in the anaesthetized guinea-pig, which was also potentiated dose-dependently by gallamine (0.1-10 mumol kg-1). 4. Gallamine (10 mumol kg-1) potentiated the bronchial anaphylactic response induced by aerosol challenge with ovalbumin in actively sensitized guinea-pigs. 5. These results suggest that neither sensory neuropeptides nor histamine are involved in the gallamine-induced potentiation of neurally-mediated bronchoconstriction, while inhibition of the sympathetic nervous system may play a minor role. They are in general agreement with the hypothesis that gallamine antagonizes acetylcholine selectively at prejunctional muscarinic receptors in the guinea-pig airways, thus increasing its release from parasympathetic nerve terminals. These autoreceptors appear to be operant during anaphylactic bronchoconstriction.

Beta-adrenoceptor desensitization induced by antigen challenge in guinea-pig trachea.

As beta-adrenoceptor function in the lung could be relevant in asthma, we carried out a functional and biochemical study of the possible occurrence of beta-receptor desensitization after the anaphylactic reaction induced in vitro in actively sensitized guinea-pig tracheas. The relaxing effect of epinephrine and vasoactive intestinal polypeptide (VIP) was tested in tracheal strips. Binding was studied with tracheal membranes and 125I-cyanopindolol. Antigen challenge resulted in a marked decrease of epinephrine-induced relaxation paralleled by a 50% reduction in beta-receptor number. The adrenergic system was specifically affected since VIP-induced relaxation was not modified by the anaphylactic reaction. In some experiments tissues were pretreated with hydrocortisone or indomethacin. Both these drugs prevented antigen exposure from impairing epinephrine relaxation, suggesting the involvement of eicosanoids in this phenomenon. Our data clearly indicate the occurrence of beta-receptor desensitization as a consequence of the anaphylactic reaction, thus impairment of the adrenergic system might play a role in asthma.