Non-adrenergic Non-cholinergic Mechanisms Research Articles

Innovative pharmacotherapies for women with overactive bladder: where are we now and what is in the pipeline?

The impressive prevalence of overactive bladder (OAB) and the relevant limitations of current treatments urge the need for novel therapeutic approaches. A systematic literature and web search was performed to identify investigational drugs that entered the early and late phases of clinical development for women with OAB symptoms. Approved pharmacological therapies for OAB (antimuscarinics, beta-3 agonists, and botulinum toxin) are evolving with the development of alternative administration methods, combination strategies, and novel compounds, expected to improve effectiveness, bladder selectivity, and dose flexibility. A wealth of investigational compounds, developed with both public and companies' indoor nonclinical disease-oriented studies, entered the early and late stages of clinical development in the last decade. Most non-anticholinergic compounds in ongoing clinical trials target central and peripheral neurotransmitter receptors involved in neurological modulation of micturition, nonadrenergic-noncholinergic mechanisms, cyclic nucleotide metabolism, different subtypes of ion channels or peripheral receptors of prostaglandins, vanilloids, vitamin D3, and opioids. Fascinating advances are ongoing also in the field of genetic therapy. New pharmaceutical formulations and drug combinations are expected to be available in the next decade in order to overcome the limitations of current drugs for OAB. Although proof-of-concept, patient-oriented studies yielded disappointing results for several tentative drugs, a lot of clinical research is ongoing that is expected to provide clinicians with novel therapeutic agents in the near future.

Excitatory effect of morphine and opioid peptides in the rat isolated colon.

Morphine and the opioid peptides cause isolated segments of rat colon to contract and relax rhythmically. This study re-examines two hypotheses to explain this phenomenon: Release of 5-hydroxytryptamine (5-HT)/acetylcholine by morphine or inhibition of a tonically active non-adrenergic, non-cholinergic (NANC) inhibitory mechanism. Rhythmic contractions induced by morphine (5 x 10(-6) M) were naloxone sensitive (10(-6) M) but unaffected by methysergide (10(-6) M), atropine (10(-6) M) or pretreatment of rats with p-chlorophenylalanine (200 mg kg-1 i.p. for four days) which lowered the 5-HT level in the colon from 3.73 +/- 0.83 mg g-1 in controls to 0.41 +/- 0.06 mg g-1 (P less than 0.001). The pattern of rhythmic contractions produced by morphine was unlike those produced by 5-HT (5 x 10(-6) M), acetylcholine (5 x 10(-6) M) or potassium chloride (30 mM). Tetrodotoxin (10(-6) M), apamin (10(-8) M), clonidine (2 x 10(-8) M), phentolamine (10(-5) M) or oxprenolol (10(-5) M) caused rhythmic contractions which were unaffected by naloxone. Clonidine contractions were inhibited by yohimbine (10(-7) M) but not by prazosin (10(-6) M). Electrical field stimulation at the peak of a contraction induced by morphine, apamin or clonidine, produced an inhibitory response which was unaffected by atropine, phentolamine, propranolol and guanethidine (all 10(-5) M). It persisted in colon segments from the rats with reserpine or 6-hydroxydopamine. These results suggest that neither the 5-HT/acetylcholine hypothesis nor inhibition of the NANC mechanism adequately explains the excitatory effect of morphine in the rat colon.

Investigational Drug Therapies for Overactive Bladder Syndrome: The Potential Alternatives to Anticolinergics

Background Overactive bladder is a high prevalent and quality of life affecting disease. The mainstay of the medical therapy is represented by antimuscarinic drugs, but their side effects markedly affect patient compliance and prompt studies on novel investigational drugs. Methods A systematic literature search of peer-reviewed papers and meeting abstracts published by December 2008 was performed. PubMed databank was searched for original English articles, by using the following search terms: “overactive bladder” or “detrusor overactivity” or “urinary incontinence” and “treatment”, alone and linked to any potential molecular target or novel drug cited in the literature. Results Effective alternative pharmacological treatments are currently scarce, but many new promising compounds are emerging which target key molecular pathways involved in micturition control. The most promising potential therapeutic targets include central nervous system GABAergic inhibitory pathway, dopaminergic and serotoninergic systems, b-adrenoceptors and cAMP metabolism, nonadrenergic-noncholinergic mechanisms such as purinergic and neuropeptidergic systems, vanilloid receptor, bladder sensory nervous terminals, nonneuronal bladder signalling systems including urothelium and interstitial cells, prostanoids, Rho-kinase and different subtypes of potassium and calcium channels. Conclusions Despite the enormous amount of new biologic insight, very few novel pharmacological therapies seems to have passed the proof-of-concept clinical stage. The ultimate clinical utility of new drugs will depend on the ability to exploit tissue-specific differences and disease-related changes in molecular expression/function and to improve storage phase dysfunctions without interfering with the emptying phase. Further preclinical investigations and controlled clinical trials are urgently needed in this challenging field.

Mechanism for the Inhibition of Contractile Activity of the Gastric Antrum and Pylorus in Rabbits during Psychogenic Stress

Psychogenic stress in rabbits (fixation to a frame) was accompanied by the inhibition of contractile activity of the gastric antrum and pylorus. These changes persisted during blockade of muscarinic receptors, nicotinic receptors, alpha(2)-adrenoceptors, and beta(1)/beta(2) adrenoceptors. A stress-induced decrease in gastric motor activity was mediated by the nonadrenergic noncholinergic mechanism. It resulted from the influence of a hormonal stress factor on the stomach, which was probably realized through nonadrenergic inhibitory neurons of the enteric nervous system.

Emerging pharmacological targets in overactive bladder therapy: experimental and clinical evidences

Antimuscarinics are the mainstay of the medical therapy for overactive bladder, but their side effects and often modest success have prompted studies on novel pharmacological approaches. In this paper, we give a systematic literature review of peer-reviewed papers on the subject. Effective nonantimuscarinic treatments are currently scarce, but many new promising compounds are emerging, which target key molecular pathways involved in micturition control. The most promising potential therapeutic targets include: nervous GABAergic, glycinergic, dopaminergic, and serotonergic systems; b-adrenoceptors and cAMP metabolism; nonadrenergic-noncholinergic mechanisms such as purinergic and neuropeptidergic systems; vanilloid receptors; bladder afferent nerves; nonneuronal bladder signaling systems including urothelium and interstitial cells; prostanoids; Rho-kinase; and different subtypes of potassium and calcium channels. Despite the enormous amount of new biologic insight, very few drugs with mechanism of action other than antimuscarinics have passed as yet the proof-of-concept stage. Further preclinical and clinical studies are urgently needed in this rapidly moving field.

Phentolamine relaxes human corpus cavernosum by a nonadrenergic mechanism activating ATP-sensitive K+ channel

To investigate the pharmacodynamics of phentolamine in human corpus cavernosum (HCC) with special attention to the role of the K+ channels. Strips of HCC precontracted with nonadrenergic stimuli and kept in isometric organ bath immersed in a modified Krebs-Henseleit solution enriched with guanethidine and indomethacine were used in order to study the mechanism of the phentolamine-induced relaxation. Phentolamine caused relaxation (approximately 50%) in HCC strips precontracted with K+ 40 mM. This effect was not blocked by tetrodotoxin (1 microM) (54.6+/-4.6 vs 48.9+/-6.4%) or (atropine (10 microM) (52.7+/-6.5 vs 58.6+/-5.6%). However, this relaxation was significantly attenuated by L-NAME (100 microM) (59.7+/-5.8 vs 27.8+/-7.1%; P<0.05; n = 8) and ODQ (100 microM) (62.7+/-5.1 vs 26.8+/-3.9%; P<0.05; n = 8). Charybdotoxin and apamin (K(Ca)-channel blockers) did not affect the phentolamine relaxations (54.6+/-4.6 vs 59.3+/-5.2%). Glibenclamide (100 microM), an inhibitor of K(ATP)-channel, caused a significant inhibition (56.7+/-6.3 vs 11.3+/-2.3%; P<0.05; n = 8) of the phentolamine-induced relaxation. In addition, the association of glibenclamide and L-NAME almost abolished the phentolamine-mediated relaxation (54.6+/-5.6 vs 5.7+/-1.4%; P<0.05; n = 8). The results suggest that phentolamine relaxes HCC by a nonadrenergic-noncholinergic mechanism dependent on nitric oxide synthase activity and activation of K(ATP)-channel.

Involvement of nonadrenergic noncholinergic mechanisms in bronchomotor response to capsaicin and electrical field stimulation in immature guinea pigs

Involvement of nonadrenergic noncholinergic mechanisms in bronchomotor response to capsaicin and electrical field stimulation in immature guinea pigs

Influence of parasympathetic non-adrenergic, non-cholinergic mechanisms on [ 3H]thymidine uptake in rat parotid salivary glands in response to mastication following upon a liquid regimen

A role for parasympathetic non-adrenergic, non-cholinergic (NANC) mechanisms in parotid gland cell hyperplasia in response to mastication was investigated. Following a liquid regimen for 1 week, aiming at glandular inactivity, rats were offered hard chow for 2 h to throw their parotid glands into secretory activity. In response to the increasing demands, [ 3H]thymidine incorporation into the trichloroacetic acid-insoluble material increased by 76% in normally innervated glands, while there was no increase in glands parasympathetically denervated in advance. In innervated glands of rats pretreated with atropine and α- and β-adrenoceptor blockers the increase was 46%. The results suggest a potential contribution of parasympathetic NANC mechanisms to the mitotic response of the parotid gland under natural feeding conditions.

Gustatory-salivary reflexes induce non-adrenergic, non-cholinergic acinar degranulation in the rat parotid gland.

In the presence of the muscarinic blocker atropine, the alpha-adrenoceptor blocker phentolamine and the beta-adrenoceptor blocker propranolol (2 mg kg(-1) of each, I.P.), the numerical density of parotid acinar secretory granules was reduced by 32 % in response to ascorbic acid (0.5 M) applied on the tongue every 30 s over 30 min in awake rats. This non-adrenergic, non-cholinergic (NANC) response was entirely dependent on an intact auriculo-temporal nerve supply. The NANC mechanisms were found to be potentially responsible for almost all of the exocytotic response that occurs in the absence of the three autonomic receptor blockers. No sympathetic contribution to the exocytotic response was found and furthermore, studies in parasympathetically denervated glands showed that the sympathetic contribution to the salivary flow was small. Experimental Physiology (2001) 86.4, 475-480.

Non-adrenergic, non-cholinergic reflex secretion of parotid saliva in rats elicited by mastication and acid applied on the tongue.

Chewing and acid on the tongue evoked a flow of saliva from the duct-cannulated parotid gland of the conscious rat despite pretreatment with atropine and adrenoceptor antagonists. This non-adrenergic, non-cholinergic (NANC) response depended on an intact parasympathetic innervation and was abolished by a tachykinin antagonist. The present findings are consistent with a physiological role for the secretory NANC mechanisms of the salivary glands.

Parasympathetic non-adrenergic, non-cholinergic reflex secretion of parotid acinar granules in rats pretreated with atropine and adrenoceptor antagonists

In parotid glands of fasted rats, feeding for 60–90 min reduced numerical density of acinar secretory granules by 52% and total activity of glandular amylase by 38%. In the presence of adrenoceptor antagonists (phentolamine and propranolol, 2 mg/kg, i.p., of each) the corresponding decrease was 31% and 32%, and in combination with atropine (1 mg/kg, i.p.) 32% as to both these indices. These non-adrenergic non-cholinergic (NANC) responses depended on an intact parasympathetic auriculo-temporal nerve since they did not occur following section of this nerve. In anaesthetized animals, electrical stimulation of the sympathetic innervation (50 Hz, 1 s every 10th second) for 60 min caused secretion of parotid saliva and decreases in number of acinar granules, by 65%, and in total amylase activity, by 49%; the adrenoceptor antagonists at the above-mentioned doses completely abolished the expected responses of the parotid gland and thus showing that these antagonists at the dose level chosen achieved an effective receptor blockade. In conclusion, parasympathetic NANC mechanisms are reflexly mobilized in response to eating, and these mechanisms are potentially responsible for the whole parasympathetic response and for the major part of that response in the absence of any antagonists. Under natural reflex conditions adrenergic, cholinergic and non-adrenergic, non-cholinergic mechanisms are most likely engaged to varying extents in order to generate the most purposeful response.

Control of the human ileocaecal junction: an in vitro analysis of adrenergic and non-adrenergic non-cholinergic mechanisms.

Circular muscle strips from the human ileocaecal junction developed myogenic tone (sustained resting tension) in vitro and showed an obvious component of relaxation in response to electrical field stimulation. These features are considered typical of sphincteric muscle because circular muscle strips from terminal ileum 1-2 cm proximal to the junction and from the caecum or ascending colon 1-2 cm distal to the junction developed little or no tone and showed predominantly contractile responses to electrical-field stimulation. In 18 out of 29 specimens, junctional strips showed excitatory responses to adrenaline or noradrenaline. The relaxation of junctional strips was non-adrenergic non-cholinergic (NANC) in nature. Nitric oxide appears to be involved in the NANC relaxation.

Contribution of neural intrapancreatic non-cholinergic non-adrenergic mechanisms to glucose-induced insulin release in the isolated rat pancreas.

In the isolated rat pancreas the effect of intrapancreatic non-adrenergic non-cholinergic nerves was examined upon insulin, glucagon and somatostatin release during perturbations of perfusate glucose. Elevation of glucose from 1.6 to 8.3 mmol/l increased insulin and somatostatin secretion and inhibited glucagon release. The first phase of insulin secretion was significantly reduced by the neurotoxin tetrodotoxin to 55% of the controls (p < 0.05). The somatostatin response was attenuated by tetrodotoxin while the change of glucagon remained unaffected. In contrast the combined adrenergic and cholinergic blockade with atropine, phentolamine and propranolol (10(-5) mol/l) did not modify the insulin, glucagon and somatostatin response. When glucose was changed from 8.3 to 1.6 mmol/l, the reduction of insulin and somatostatin release was not modified by tetrodotoxin, but stimulation of glucagon was significantly attenuated by 60-70% (p < 0.03), which was similar to the effect of combined adrenergic and cholinergic blockade. Subsequently, the effect of neural blockade was examined during more physiological perturbations of perfusate glucose levels. When glucose was changed from 3.9 to 7.2 mmol/l, tetrodotoxin also attenuated first phase insulin response by 40% while cholinergic and adrenergic blockade had no effect. The nitric oxide synthase inhibitor NG-Nitro-L-arginine-methyl-ester (L-NAME) did not alter the glucose-induced insulin response indicating that nitric oxide is not involved in this mechanism. It is concluded that neural non-adrenergic non-cholinergic mechanisms contribute to the first, but not second phase of glucose-induced insulin release. Non-adrenergic non-cholinergic effects do not participate in regulation of glucagon and somatostatin secretion under the conditions employed.(ABSTRACT TRUNCATED AT 250 WORDS)

NON‐ADRENERGIC, NON‐CHOLINERGIC NEURAL CONTROL OF THE AIRWAYS

1. In addition to the classical cholinergic bronchoconstrictor and adrenergic bronchodilator neural mechanisms, there is a large volume of evidence to suggest the existence of neural pathways within the airways of a variety of species which are neither adrenergic nor cholinergic, the non-adrenergic, non-cholinergic (NANC) mechanisms. With respect to airway smooth muscle tone, NANC neural responses may induce either contraction (excitatory, e-NANC) or relaxation (inhibitory, i-NANC). Early investigations of NANC mechanisms in both human and other animal airways suggested a role for neuropeptides as the putative neurotransmitters. 2. Excitatory NANC (e-NANC) bronchoconstrictor responses are believed to be mediated by the release of sensory neuropeptides from a subpopulation of non-myelinated C-fibre primary afferent neurones in the airways. e-NANC nerves, which release tachykinins such as substance P (SP), neurokinin A (NKA) and the peptide calcitonin gene-related peptide (CGRP, produced as a result of alternative splicing of the calcitonin gene) are selectively degenerated by the nerve toxin capsaicin (an extract from hot peppers), with the subsequent abolition of the e-NANC responses. Tachykinin receptors have been detected by radio-ligand receptor binding studies and visualized by autoradiographic mapping, and exogenous addition of these peptides elicits a bronchoconstrictor response in both human and other animal airways. In addition to these effects on airway smooth muscle tone, tachykinins produce an increase in microvascular permeability (and associated oedema formation), mucus hypersecretion and cause an exaggerated cholinergic bronchoconstrictor response. Thus, tachykinins may play a role in the inflammatory process and contribute to the neurogenic inflammation as seen in asthma.(ABSTRACT TRUNCATED AT 250 WORDS)

Control of the bronchial tone.

The tone of airway smooth muscle is the functional expression of a dynamic equilibrium between various excitatory and inhibitory mechanisms. On the excitatory side, the vagal reflex mechanism, mainly activated by central airway irritant receptors, is the most important. Bronchial epithelium plays a pivotal role in the regulation of the responsiveness of these receptors. Cholinergic responses of the bronchial muscle are mediated by muscarinic (M2) receptors. On the inhibitory side, the main mechanism is the adrenergic one: beta 2-receptors are mainly stimulated by circulating adrenaline. A dense population of beta 2-receptors has recently been observed in bronchial epithelium. Nonadrenergic-noncholinergic mechanisms--both excitatory and inhibitory--participate in the regulation of bronchial motor tone, but their role in man is still to be defined. Nonsteroidal anti-inflammatory drugs do not modify bronchial motor tone; thus, in this respect, at least in physiological conditions, a modulatory function of the prostaglandin system can be excluded. Probably the same applies to leukotrienes, but for a definite answer, specific antagonists are needed.

Autonomic innervation of pulmonary artery: evidence for a nonadrenergic noncholinergic inhibitory system.

We have investigated the presence and significance of the nonadrenergic noncholinergic relaxant system in isolated strips of guinea pig and cat pulmonary artery (PA), using transmural electrical field stimulation (EFS). Changes in PA strip tension were measured isometrically, in response to EFS with 50-70 V at 20 Hz and pulse durations of 1 or 2 ms. We examined the influence on these responses of alpha- and beta-adrenergic and cholinergic muscarinic receptor blockade, the neurotoxin tetrodotoxin, and the local anesthetic lidocaine. EFS induced an initial brief contraction followed by a slow prolonged relaxation. Phenoxybenzamine, 1.5 X 10(-5) M, greatly reduced the contraction (74.1-87.6% in guinea pig, 65.7-67.4% in cat) and enhanced the subsequent relaxation (23.9-35.5% in guinea pig, 66.8-67.8% in cat). The relaxation was moderately reduced by propranolol, 7.7 X 10(-6) M (30.5-35.0% in guinea pigs, 6.0-21.2% in cat), and by atropine, 1.4 X 10(-6) M (23.7-32.1% in guinea pig, 24.9-40.6% in cat). The major portion of the relaxation, however, persisted in the presence of these blockers. The neurogenic origin of this relaxation was confirmed by its loss with either tetrodotoxin or lidocaine. We conclude that in guinea pig and cat lungs (1) neurogenic pulmonary vascular contraction is principally mediated by the alpha-adrenergic system, (2) the beta-adrenergic and cholinergic components have a moderate relaxant influence, but (3) the dominant part of neurogenic relaxation is mediated by a nonadrenergic noncholinergic mechanism.

Vagally mediated gastric relaxation induced by intestinal nutrients in the dog.

Using a gastric barostat to measure gastric tone, we previously demonstrated that nutrient perfusion into the intestine induces gastric relaxation. To investigate the pathway of this enterogastric reflex we surgically isolated the vagi either in a cervical skin tunnel (3 dogs) or within an implanted supradiaphragmatic cooling jacket (3 dogs). In the conscious fasted dogs, cervical or supradiaphragmatic vagal blockade by cooling (5 degrees C X 10 min) induced a reversible gastric relaxation. Bethanechol (0.2 mg X kg-1 X h-1 iv) alone or in combination with adrenergic blockers (phentolamine 1.5 mg X kg-1 X h-1 + propranolol 0.3 mg X kg-1 X h-1) suppressed the cooling-induced relaxation but did not abolish gastric relaxation induced by intestinal nutrient perfusion (Osmolite, 3.1 ml/min). At this point, vagal cooling, either cervical or supradiaphragmatic, reversibly blocked the nutrient-induced gastric relaxation: gastric tone significantly increased driven by the cholinergic background and reverted after vagal rewarming to the previous relaxed state. We conclude that intestinal nutrients induce gastric relaxation by a nonadrenergic noncholinergic mechanism. This reflex is mediated by fibers contained in the vagus nerves at both cervical and supradiaphragmatic levels.

Capsaicin activates neurogenic non-adrenergic non-cholinergic relaxations of the isolated rat duodenum

Capsaicin induces a transient relaxation of the isolated rat duodenum in the presence of atropine plus guanethidine. The relaxant effect of capsaicin was not observed in vitro in the duodenum of capsaicin-pretreated animals. Capsaicin-induced relaxation was antagonized by tetrodotoxin or cold storage (24 h at 4°C). It is concluded that capsaicin produces a neurogenic relaxation of the rat duodenum by activating an intramural non-adrenergic non-cholinergic mechanism which inhibits intestinal motility.

The effect of hypoxia on neuroeffector transmission in the bovine retractor penis and rat anococcygeus muscles

The effects of reducing the PO2 of the bathing fluid were studied on non-adrenergic non-cholinergic (NANC) transmission in isolated preparations of the bovine retractor penis muscle, the rat anococcygeus muscle, the guinea-pig taenia caeci and the guinea-pig urinary bladder. Hypoxia rapidly and reversibly impaired NANC transmission in the bovine retractor penis and rat anococcygeus muscles but did not affect transmission in the guinea-pig taenia caeci or bladder, suggesting that different NANC mechanisms are involved. Although neurally-evoked relaxation of the bovine retractor penis was impaired by hypoxia, relaxations produced by vasoactive intestinal peptide, prostaglandin E1, sodium nitroprusside or an inhibitory factor isolated from the bovine retractor penis were unaffected. Since the inhibitory factor is similar to, and may actually be the NANC transmitter, the results suggest that the site of action of hypoxia in impairing transmission is prejunctional at the inhibitory nerve endings.

The internal anal sphincter in the cat: a study of nervous mechanisms affecting tone and reflex activity.

1. Smooth muscle activities in rectum and internal anal sphincter have been recorded using intraluminal balloons.2. Reflex activation of the sphincter, caused by distension of the rectum, has been assessed before and after various combinations of blocking drugs.3. Responses to stimulation of hypogastric or sacral nerves, or to the administration of drugs with autonomic actions have been tested before and after various combinations of blocking drugs.4. Results indicate that the tone of the internal anal sphincter is influenced by a number of neural mechanisms. These include motor pathways involving both alpha-adrenergic and cholinergic mechanisms and inhibitory pathways involving both beta-adrenergic and non-adrenergic non-cholinergic mechanisms.5. Cholinergic contractions of the sphincter were converted to relaxations after alpha-adrenergic blockade. This indicates that the contractions are an indirect effect operating through an adrenergic reflex. Cholinergic relaxations may also be indirect and operate through reflex inhibition secondary to rectal contractions.6. Sphincteric motor activity is controlled largely through alpha-adrenergic mechanisms by adrenergic nerves acting directly on the muscle. beta-Adrenergic inhibitory mechanisms are thought to operate indirectly via ganglia.7. The over-all control of the sphincter is by complex reflex mechanisms involving numerous pathways and the activity of the sphincter at any one time is determined by the net balance between motor and inhibitory influences.8. Sacral nerve stimulation indicated that it contains cholinergic nerves to the rectum, non-adrenergic non-cholinergic inhibitory axons to the sphincter and variable numbers of adrenergic axons to the sphincter.9. Responses of the sphincter to drugs and nerve stimulation were often variable, as has been described many times in the literature. It is considered that this is due to complex combinations of indirect reflex effects, secondary to activation of structures outside the sphincter, operating with or against direct effects on the sphincter itself.