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1.
Eur J Pharmacol ; 945: 175603, 2023 Apr 15.
Article in English | MEDLINE | ID: mdl-36804548

ABSTRACT

The motor activity of the epididymis duct is an essential process for male fertility and it is regulated by hormonal, neuronal and epithelial mechanisms. However, although there is evidence for the presence of histamine in the epididymis, its effects on epididymal motor activity are unknown. This study sought to evaluate the contractile effects of histamine on the rat distal cauda epididymis duct. Segments of the distal cauda epididymis duct from male Wistar rats were isolated and used in isolated organ bath experiments to evaluate the contractile effects of histamine in the absence or presence of antagonists of histamine receptors, α1-adrenoceptors and muscarinic acetylcholine receptors. The effects of histamine on noradrenaline induced contractions were also investigated. Histamine was able to induce phasic contractions on rat distal cauda epididymis duct which were prevented by promethazine 10-1000 nM (H1 receptor antagonist), ranitidine 1-100 µM (H2 receptor antagonist), atropine 100 nM (muscarinic antagonist), and prazosin 100 nM (α1-adrenoceptor antagonist). In addition, histamine was also able to modify noradrenaline-induced contractions possibly via activation of H1 and H2 receptors. In conclusion, this study demonstrates that histamine can induce phasic contractions of rat distal cauda epididymis via H2 receptors and autonomic neurotransmitters. Histamine may also exert modulatory actions on contractions of rat distal cauda epididymis duct induced by adrenergic receptor agonists. Further studies are necessary to unveil the localization of histamine receptors within the epididymal duct and the consequences of manipulation of the histaminergic system on epididymal function and male fertility.


Subject(s)
Epididymis , Histamine , Rats , Male , Animals , Rats, Wistar , Histamine/pharmacology , Prazosin/pharmacology , Norepinephrine/pharmacology , Receptors, Adrenergic, alpha-1
2.
Purinergic Signal ; 18(4): 421-433, 2022 12.
Article in English | MEDLINE | ID: mdl-35821454

ABSTRACT

Within the family of purinergic receptors, the P2X1 receptor is a ligand-gated ion channel that plays a role in urogenital, immune and cardiovascular function. Specifically, the P2X1 receptor has been implicated in controlling smooth muscle contractions of the vas deferens and therefore has emerged as an exciting drug target for male contraception. In addition, the P2X1 receptor contributes to smooth muscle contractions of the bladder and is a target to treat bladder dysfunction. Finally, platelets and neutrophils have populations of P2X1 receptors that could be targeted for thrombosis and inflammatory conditions. Drugs that specifically target the P2X1 receptor have been challenging to develop, and only recently have small molecule antagonists of the P2X1 receptor been available. However, these ligands need further biological validation for appropriate selectivity and drug-like properties before they will be suitable for use in preclinical models of disease. Although the atomic structure of the P2X1 receptor has yet to be determined, the recent discovery of several other P2X receptor structures and improvements in the field of structural biology suggests that this is now a distinct possibility. Such efforts may significantly improve drug discovery efforts at the P2X1 receptor.


Subject(s)
Receptors, Purinergic P2X1 , Male , Humans , Urinary Bladder , Muscle Contraction , Vas Deferens/physiology , Blood Platelets , Receptors, Purinergic P2X , Adenosine Triphosphate
3.
PLoS One ; 17(7): e0271735, 2022.
Article in English | MEDLINE | ID: mdl-35900970

ABSTRACT

Stinging nettle root and leaf extracts were tested for their effect on prostatic smooth muscle contractility. Root extract did not affect electrical field stimulation induced-nerve mediated contractions of isolated rat prostates. On the other hand, leaf extract attenuated electrical field stimulation-induced contractions at all frequencies. Similarly, contractions elicited by exogenous administration of ATP and αß-methylene ATP were inhibited by leaf extract, whereas contractions elicited by exogenous administration of noradrenaline or acetylcholine were unaffected. The active component was present within the aqueous phase of the leaf extract. In mouse mating studies, stinging nettle leaf extract (50 mg p.o. daily) reduced male fertility by 53% compared to vehicle-treated male mice. Cardiovascular parameters were unaffected by administration of stinging nettle leaf extract (p ≥ 0.057). Treated mice exhibited normal mating behaviour. Bladder and testes weighed less in stinging nettle leaf extract treated mice. All other organs and total body weight were unaffected. It is concluded that stinging nettle leaf extract reduces contractility of genitourinary smooth muscle by acting as an antagonist at postjunctional P2X1-purinoceptors. These data indicates that blocking sperm transport through pharmacological blockade of P2X1-purinoceptors via oral administration is consistent with an effective and convenient biological strategy male contraception.


Subject(s)
Urtica dioica , Adenosine Triphosphate , Animals , Fertility , Male , Mice , Plant Extracts/pharmacology , Purinergic P2 Receptor Antagonists , Rats , Receptors, Purinergic , Receptors, Purinergic P2 , Seeds
4.
Biol Reprod ; 103(2): 323-332, 2020 08 04.
Article in English | MEDLINE | ID: mdl-32648904

ABSTRACT

Sympathetically mediated contractions of smooth muscle cells in the vasa deferentia are mediated by neuronally released adenosine 5'-triphosphate (ATP) and noradrenaline, which stimulate P2X1-purinoceptors and α1A-adrenoceptors, respectively. This process is crucial for sperm transport, as demonstrated in knockout mouse studies where simultaneous genetic deletion of P2X1-purinoceptors and α1A-adrenoceptors resulted in male infertility. We hypothesize that dual pharmacological antagonism of these two receptors could inhibit sperm transport sufficiently to provide a novel nonhormonal method of male contraception. To generate a suitable P2X1-purinoceptor antagonist, substituents were introduced on the phenyl moiety of 2-phenyl-5,6,7,8-tetrahydroquinoxaline to create a series of analogues that were tested for P2X1-purinoceptor antagonism in isolated preparations of rat vas deferens. Novel compounds were initially screened for their ability to attenuate contractile responses to electrical field stimulation (EFS: 60 V, 0.5 ms, 0.2 Hz). The addition of polar substituents to the meta, but not ortho, position markedly increased the inhibition of contractions, as did the addition of both polar and aliphatic substituents to the para position. Di-substituted compounds were also synthesized and tested, resulting in a compound 31 (2-hydroxy, 4-fluoro), which exhibited the greatest potency, with an IC50 of 14 µM (95% confidence limits: 12-16 µM). Additionally, compound 31 noncompetitively antagonized contractions mediated by exogenously administered αß-methylene ATP (10 nM-30 µM) but had no inhibitory effect on contractions mediated by exogenously administered noradrenaline (30 nM-100 µM) or acetylcholine (30 nM-100 µM). These results have contributed to a structure-activity relationship profile for the P2X1-purinoceptor that will inform future designs of more potent antagonists.


Subject(s)
Contraceptive Agents, Male , Indolizines/chemistry , Purinergic P2X Receptor Antagonists/pharmacology , Vas Deferens/drug effects , Animals , Male , Muscle Contraction/drug effects , Muscle, Smooth/drug effects , Rats , Receptors, Purinergic P2X1/metabolism , Translational Research, Biomedical
5.
Br J Pharmacol ; 176(14): 2358-2365, 2019 07.
Article in English | MEDLINE | ID: mdl-30719698

ABSTRACT

The α1A -adrenoceptor is abundantly expressed in the lower urinary tract and is the principal therapeutic target for the symptomatic treatment of lower urinary tract symptoms in men. Prazosin has a lower affinity for the lower urinary tract α1A -adrenoceptor than α1A -adrenoceptors found in other parts of the body. This has led to the lower urinary tract α1A -adrenoceptor being subclassified as an α1L -adrenoceptor. It was demonstrated that this pharmacologically distinct α1L -adrenoceptor is a product of the α1A -adrenoceptor gene, but the mechanism by which this altered phenotype is achieved remains a mystery. Hypotheses for this altered pharmacology include the presence of an interacting protein such as cysteine-rich with EGF-like domain (CRELD) 1 or other GPCRs such as the CXCR2 chemokine or 5-HT1B receptor. Alternatively, the influence of breast cancer resistance protein (BCRP) efflux transporters on the pharmacology of α1A -adrenoceptors has also been investigated. These and other hypotheses will be described and discussed in this review. LINKED ARTICLES: This article is part of a themed section on Adrenoceptors-New Roles for Old Players. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v176.14/issuetoc.


Subject(s)
Receptors, Adrenergic, alpha-1/genetics , Animals , Humans , Phenotype
6.
Article in English | MEDLINE | ID: mdl-30690282

ABSTRACT

In the present study, the anthelmintic activity of a human tyrosine kinase inhibitor, AG-1295, and 14 related tetrahydroquinoxaline analogues against Haemonchus contortus was explored. These compounds were screened against parasitic larvae - exsheathed third-stage (xL3) and fourth-stage (L4) - using a whole-organism screening assay. All compounds were shown to have inhibitory effects on larval motility, development and growth, and induced evisceration through the excretory pore in xL3s. The estimated IC50 values ranged from 3.5 to 52.0 µM for inhibition of larval motility or development. Cytotoxicity IC50 against human MCF10A cells was generally higher than 50 µM. Microscopic studies revealed that this eviscerated (Evi) phenotype occurs rapidly (<20 min) and relates to a protrusion of internal tissues and organs (evisceration) through the excretory pore in xL3s; severe pathological damage in L4s as well as a suppression of larval growth in both stages were also observed. Using a relatively low concentration (12.5 µM) of compound m10, it was established that the inhibitor has to be present for a relatively short time (between 30 h and 42 h) during in vitro development from xL3 to L4, to induce the Evi phenotype. Increasing external osmotic pressure prevented evisceration and moulting, and xL3s remained unaffected by the test compound. These results point to a mode of action involving a dysregulation of morphogenetic processes during a critical time-frame, in agreement with the expected behaviour of a tyrosine kinase inhibitor, and suggest potential for development of this compound class as nematocidal drugs.


Subject(s)
Antinematodal Agents/pharmacology , Haemonchus/drug effects , Quinoxalines/pharmacology , Tyrphostins/pharmacology , Animals , Biological Assay , Drug Discovery , Haemonchus/physiology , Inhibitory Concentration 50 , Larva/drug effects , Larva/physiology , Molting/drug effects , Phenotype
8.
Br J Pharmacol ; 174(14): 2318-2333, 2017 Jul.
Article in English | MEDLINE | ID: mdl-28444738

ABSTRACT

BACKGROUND AND PURPOSE: Agonists acting at GPCRs promote biased signalling via Gα or Gßγ subunits, GPCR kinases and ß-arrestins. Since the demonstration of biased agonism has implications for drug discovery, it is essential to consider confounding factors contributing to bias. We have examined bias at human α1A -adrenoceptors stably expressed at low levels in CHO-K1 cells, identifying off-target effects at endogenous receptors that contribute to ERK1/2 phosphorylation in response to the agonist oxymetazoline. EXPERIMENTAL APPROACH: Intracellular Ca2+ mobilization was monitored in a Flexstation® using Fluo 4-AM. The accumulation of cAMP and ERK1/2 phosphorylation were measured using AlphaScreen® proximity assays, and mRNA expression was measured by RT-qPCR. Ligand bias was determined using the operational model of agonism. KEY RESULTS: Noradrenaline, phenylephrine, methoxamine and A61603 increased Ca2+ mobilization, cAMP accumulation and ERK1/2 phosphorylation. However, oxymetazoline showed low efficacy for Ca+2 mobilization, no effect on cAMP generation and high efficacy for ERK1/2 phosphorylation. The apparent functional selectivity of oxymetazoline towards ERK1/2 was related to off-target effects at 5-HT1B receptors endogenously expressed in CHO-K1 cells. Phenylephrine and methoxamine showed genuine bias towards ERK1/2 phosphorylation compared to Ca2+ and cAMP pathways, whereas A61603 displayed bias towards cAMP accumulation compared to ERK1/2 phosphorylation. CONCLUSION AND IMPLICATIONS: We have shown that while adrenergic agonists display bias at human α1A -adrenoceptors, the marked bias of oxymetazoline for ERK1/2 phosphorylation originates from off-target effects. Commonly used cell lines express a repertoire of endogenous GPCRs that may confound studies on biased agonism at recombinant receptors.


Subject(s)
Adrenergic alpha-1 Receptor Agonists/pharmacology , Imidazoles/pharmacology , Methoxamine/pharmacology , Norepinephrine/pharmacology , Phenylephrine/pharmacology , Receptors, Adrenergic, alpha-1/metabolism , Tetrahydronaphthalenes/pharmacology , Adrenergic alpha-1 Receptor Agonists/chemistry , Animals , CHO Cells , Cells, Cultured , Cricetulus , Dose-Response Relationship, Drug , Humans , Imidazoles/chemistry , Methoxamine/chemistry , Norepinephrine/chemistry , Phenylephrine/chemistry , Structure-Activity Relationship , Tetrahydronaphthalenes/chemistry
9.
Oncotarget ; 7(28): 44492-44504, 2016 Jul 12.
Article in English | MEDLINE | ID: mdl-27283984

ABSTRACT

Progression of castration-resistant tumors is frequent in prostate cancer. Current systemic treatments for castration-resistant prostate cancer only produce modest increases in survival time and self-renewing Tumor-Initiating Cells (TICs) are suspected to play an important role in resistance to these treatments. However it remains unclear whether the same TICs display both chemo-resistance and self-renewing abilities throughout progression from early stage lesions to late, castration resistant tumors. Here, we found that treatment of mice bearing LNCaP-derived xenograft tumors with cytotoxic (docetaxel) and anti-androgen (flutamide) compounds enriched for cells that express TROP2, a putative TIC marker. Consistent with a tumor-initiating role, TROP2high cells from androgen-sensitive prostate cancer cell lines displayed an enhanced ability to re-grow in culture following treatment with taxane-based chemotherapy with or without androgen blockade. TROP2 down-regulation in these cells reduced their ability to recur after treatment with docetaxel, in the presence or absence of flutamide. Accordingly, in silico analysis of published clinical data revealed that prostate cancer patients with poor prognosis exhibit significantly elevated TROP2 expression level compared to low-risk patients, particularly in the case of patients diagnosed with early stage tumors. In contrast, in androgen-independent prostate cancer cell lines, TROP2high cells did not exhibit a differential treatment response but were characterized by their high self-renewal ability. Based on these findings we propose that high TROP2 expression identifies distinct cell sub-populations in androgen-sensitive and androgen-independent prostate tumors and that it may be a predictive biomarker for prostate cancer treatment response in androgen-sensitive tumors.


Subject(s)
Antigens, Neoplasm/genetics , Cell Adhesion Molecules/genetics , Flutamide/pharmacology , Gene Expression Regulation, Neoplastic , Prostatic Neoplasms/drug therapy , Taxoids/pharmacology , Androgen Antagonists/administration & dosage , Androgen Antagonists/pharmacology , Animals , Antigens, Neoplasm/metabolism , Antineoplastic Agents/administration & dosage , Antineoplastic Agents/pharmacology , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Cell Adhesion Molecules/metabolism , Cell Survival/drug effects , Cell Survival/genetics , Docetaxel , Flutamide/administration & dosage , Gene Expression Profiling/methods , Kaplan-Meier Estimate , Male , Mice, Inbred NOD , Mice, SCID , Neoplastic Stem Cells/drug effects , Neoplastic Stem Cells/metabolism , Neoplastic Stem Cells/pathology , Prostatic Neoplasms/genetics , Prostatic Neoplasms/metabolism , Prostatic Neoplasms, Castration-Resistant/drug therapy , Prostatic Neoplasms, Castration-Resistant/genetics , Prostatic Neoplasms, Castration-Resistant/metabolism , Taxoids/administration & dosage , Xenograft Model Antitumor Assays/methods
10.
Neurourol Urodyn ; 34(5): 482-8, 2015 Jun.
Article in English | MEDLINE | ID: mdl-24436088

ABSTRACT

AIMS: To examine the effects of the α1A -adrenoceptor antagonist, tamsulosin, on spontaneous contractile and electrical activity in the guinea-pig prostate gland. METHODS: The effects of tamsulosin (0.1 and 0.3 nM) were investigated in adult and ageing male guinea pig prostate glands using conventional tension recording and electrophysiological intracellular microelectrode recording techniques. RESULTS: Tamsulosin reduced spontaneous activity, and had different age-dependent effects on adult and ageing guinea pigs at different concentrations. 0.1 nM tamsulosin caused a significantly greater reduction of spontaneous contractile and electrical activity in ageing guinea pigs in comparison to adult guinea pigs. In contrast, 0.3 nM tamsulosin had a significantly greater reduction of spontaneous contractile and electrical activity in adult guinea pigs in comparison to ageing guinea pigs. CONCLUSIONS: This study demonstrates that tamsulosin can modulate spontaneous myogenic stromal contractility and the underlying spontaneous electrical activity; tamsulosin does not block spontaneous activity. This reduction in spontaneous activity suggests that downstream cellular mechanisms underlying smooth muscle tone are being targeted, and these may represent novel therapeutic targets to better treat benign prostatic hyperplasia.


Subject(s)
Adrenergic alpha-1 Receptor Antagonists/pharmacology , Aging , Membrane Potentials/drug effects , Muscle Contraction/drug effects , Muscle, Smooth/drug effects , Prostate/drug effects , Sulfonamides/pharmacology , Animals , Electrophysiological Phenomena/drug effects , Guinea Pigs , Male , Tamsulosin
11.
Neurourol Urodyn ; 34(3): 292-8, 2015 Mar.
Article in English | MEDLINE | ID: mdl-24249481

ABSTRACT

AIMS: An age-related increase in prostatic smooth muscle tone is partly responsible for the lower urinary tract symptoms associated with benign prostatic hyperplasia (BPH). Changes in the effectors of prostatic smooth muscle contraction with age may play a role in the development of these symptoms. Using a mouse model of prostate contractility, this study investigated the effect of age on the different components of contractility in the prostate gland. METHODS: The isometric force developed in response to electrical field stimulation or exogenously applied agonists by mouse prostates mounted in organ baths, was evaluated to determine the effect of age on contractile mechanisms. Changes with age in the rate of ATP breakdown and levels of the P2rx1 gene and P2X1-purinoceptor expression in mouse prostate were measured by a modified luciferin-luciferase assay, RT-PCR and western blot, respectively. RESULTS: Nerve mediated contractile responses containing a component elicited by P2X1-purinoceptors were observed in prostates taken from aged mice, but not in prostates taken from young adult mice. Furthermore, the potency of the endogenous purinoceptor agonist ATP was 50-fold greater in aged mice, whereas the potency of its stable analogue α,ß-metATP was unchanged. An age-related decrease in ATP metabolism was also observed. CONCLUSIONS: With age, a purinergic contractile response to nerve stimulation develops in the mouse prostate gland due to a decrease in the rate of ATP breakdown. This may contribute to the increase in muscular tone observed in BPH and suggests that P2X1-purinoceptors are an additional target for the treatment of BPH.


Subject(s)
Adenosine Triphosphate/metabolism , Muscle Contraction , Prostate/physiology , Receptors, Purinergic P2X1/physiology , Age Factors , Animals , Male , Mice , Prostate/metabolism , Time Factors
12.
J Ethnopharmacol ; 152(2): 283-91, 2014 Mar 14.
Article in English | MEDLINE | ID: mdl-24463033

ABSTRACT

ETHNOPHARMACOLOGICAL RELEVANCE: Saw palmetto (Serenoa repens) was first used medicinally by native American Indians to treat urological disorders. Nowadays, saw palmetto extracts are widely used in Europe and North America to treat the urinary symptoms associated with benign prostatic hyperplasia even though its mechanisms of action are poorly understood. This study aimed to characterize the bioactive constituents of a lipid extract of saw palmetto that are able to affect contractility of the rat prostate gland. The mechanism of action will also be investigated. MATERIALS AND METHODS: A commercially available lipid extract of saw palmetto was subjected to fractionation using normal phase column chromatography. Composition of fractions was assessed by proton nuclear magnetic resonance spectroscopy ((1)H NMR) and mass spectrometry (MS). Contractile activities of these fractions were evaluated pharmacologically using isolated preparations of rat prostate gland and compared to the activity of the crude extract. RESULTS: Saw palmetto extract inhibited contractions of the rat prostate gland which were consistent with smooth muscle relaxant activity. Only the ethyl acetate fraction resulting from chromatography inhibited contractions of isolated rat prostates similarly to the inhibition produced by the crude lipid extract. Comparison with authentic samples and analysis of NMR data revealed that this bioactivity was due to the fatty acid components present in the ethyl acetate fraction. Bioassay using various pharmacological tools identified multiple contractile mechanisms which were affected by the bioactive constituents. CONCLUSION: A fatty acid component of saw palmetto extract causes inhibition of prostatic smooth muscle contractions via a non-specific mechanism.


Subject(s)
Muscle Contraction/drug effects , Plant Extracts/pharmacology , Prostate/drug effects , Serenoa/chemistry , Animals , Lipids/chemistry , Magnetic Resonance Spectroscopy , Male , Mass Spectrometry , Medicine, Traditional , Muscle, Smooth/drug effects , Muscle, Smooth/metabolism , Rats , Rats, Sprague-Dawley
13.
ChemMedChem ; 9(2): 399-410, 2014 Feb.
Article in English | MEDLINE | ID: mdl-24339417

ABSTRACT

Noscapine, a phthalideisoquinoline alkaloid derived from Papaver somniferum, is a well-known antitussive drug that has a relatively safe in vitro toxicity profile. Noscapine is also known to possess weak anticancer efficacy, and since its discovery, efforts have been made to design derivatives with improved potency. Herein, the synthesis of a series of noscapine analogues, which have been modified in the 6', 9', 1 and 7-positions, is described. In a previous study, replacement of the naturally occurring N-methyl group in the 6'-position with an N-ethylaminocarbonyl was shown to promote cell-cycle arrest and cytotoxicity against three cancer cell lines. Here, this modification has been combined with other structural changes that have previously been shown to improve anticancer activity, namely halo substitution in the 9'-position, regioselective O-demethylation to reveal a free phenol in the 7-position, and reduction of the lactone to the corresponding cyclic ether in the 1-position. The incorporation of new aryl substituents in the 9'-position was also investigated. The study identified interesting new compounds able to induce G2/M cell-cycle arrest and that possess cytotoxic activity against the human prostate carcinoma cell line PC3, the human breast adenocarcinoma cell line MCF-7, and the human pancreatic epithelioid carcinoma cell line PANC-1. In particular, the ethyl urea cyclic ether noscapinoids and a compound containing a 6'-ethylaminocarbonyl along with 9'-chloro, 7-hydroxy and lactone moieties exhibited the most promising biological activities, with EC50 values in the low micromolar range against all three cancer cell lines, and these derivatives warrant further investigation.


Subject(s)
Antineoplastic Agents, Phytogenic/chemistry , Antineoplastic Agents, Phytogenic/pharmacology , Noscapine/analogs & derivatives , Noscapine/pharmacology , Papaver/chemistry , Cell Cycle/drug effects , Cell Line, Tumor , Female , Humans , Male , Neoplasms/drug therapy , Structure-Activity Relationship
14.
Proc Natl Acad Sci U S A ; 110(51): 20825-30, 2013 Dec 17.
Article in English | MEDLINE | ID: mdl-24297884

ABSTRACT

Therapeutic targets for male contraception are associated with numerous problems due to their focus on disrupting spermatogenesis or hormonal mechanisms to produce dysfunctional sperm. Here we describe the dual genetic deletion of α1A-adrenergic G protein-coupled receptors (adrenoceptors) and P2X1-purinoceptor ligand gated ion channels in male mice, thereby blocking sympathetically mediated sperm transport through the vas deferens during the emission phase of ejaculation. This modification produced 100% infertility without effects on sexual behavior or function. Sperm taken from the cauda epididymides of double knockout mice were microscopically normal and motile. Furthermore, double knockout sperm were capable of producing normal offspring following intracytoplasmic sperm injection into wild-type ova and implantation of the fertilized eggs into foster mothers. Blood pressure and baroreflex function was reduced in double knockout mice, but no more than single knockout of α1A-adrenoceptors alone. These results suggest that this autonomic method of male contraception appears free of major physiological and behavioral side effects. In addition, they provide conclusive proof of concept that pharmacological antagonism of the P2X1-purinoceptor and α1A-adrenoceptor provides a safe and effective therapeutic target for a nonhormonal, readily reversible male contraceptive.


Subject(s)
Contraception , Infertility, Male/genetics , Receptors, Adrenergic, alpha-1 , Receptors, Purinergic P2X1 , Spermatozoa/metabolism , Adrenergic alpha-1 Receptor Antagonists/pharmacology , Animals , Baroreflex , Gene Knockdown Techniques , Male , Mice , Mice, Knockout , Purinergic P2X Receptor Antagonists/pharmacology , Sperm Motility/physiology , Spermatozoa/cytology
15.
Asian J Androl ; 15(6): 715-6, 2013 Nov.
Article in English | MEDLINE | ID: mdl-24141535

ABSTRACT

In the July 12 issue of Science magazine, researchers from the Albert Einstein College of Medicine, the Mount Sinai School of Medicine, the Durham VA Medical Centre and Duke University published an elegant study demonstrating that the sympathetic nervous system, acting through ß2 and ß3-adrenoceptors in the prostate, plays an important role in the initiation of prostate cancer, while the parasympathetic nervous system plays a role in the dissemination of tumour metastases via M1 muscarinic receptors. These findings are significant because they indicate that receptors associated with the autonomic nervous system may be viable targets for prostate cancer therapy.


Subject(s)
Adenocarcinoma/pathology , Autonomic Nervous System/growth & development , Neurogenesis , Prostate/innervation , Prostate/pathology , Prostatic Neoplasms/pathology , Animals , Humans , Male
16.
Eur J Pharmacol ; 721(1-3): 313-21, 2013 Dec 05.
Article in English | MEDLINE | ID: mdl-24055190

ABSTRACT

With age an increase in prostatic smooth muscle tone mediated by α1L-adrenoceptors contributes to the lower urinary tract symptoms associated with benign prostatic hyperplasia. Current treatments for benign prostatic hyperplasia include α1-adrenoceptor antagonists which inhibit smooth muscle contraction. However, muscarinic receptors also mediate prostatic smooth muscle contraction and targeting a convergent signalling pathway may be a more effective treatment strategy. This study determined signalling pathways involved in contraction by measuring isometric force developed by prostates from wild type, α1A-adrenoceptor and M3-muscarinic receptor knockout mice mounted in organ baths in response to, electrical field stimulation or exogenously applied agonists, in the presence or absence of signalling pathway inhibitors. Fluorescence immunohistochemistry was also used to confirm functional observations. Contractile responses mediated by carbachol were reduced by inhibitors of phospholipase C (U73122; 3-10 µM), L-type Ca(2+) channels (nifedipine; 1 µM), Rho kinase (Y-27632; 10-30 µM), but not protein kinase C (GF109203 X;10 µM). Nifedipine (1 µM), Y-27632 (10 µM), and GF109203 X (10 µM) inhibited nerve mediated contractile responses. Y-27632 (10-30 µM) inhibited noradrenaline mediated contractions. RhoA and ROCK2 were found to be immunolocalised with prostatic smooth-muscle. Contractions mediated by M3-muscarinic receptors in the mouse prostate involve the prototypical phospholipase C signalling pathway, as well as L-type Ca(2+) channels. Adrenergic and cholinergic components of smooth muscle contraction in the mouse prostate both involve the activation of the Rho-kinase pathway, which may be a suitable common pathway for more effective treatments of symptoms associated with benign prostatic hyperplasia.


Subject(s)
Acetylcholine/metabolism , Muscle Contraction , Muscle, Smooth/physiology , Norepinephrine/metabolism , Prostate/physiology , rho-Associated Kinases/metabolism , Animals , Enzyme Activation , In Vitro Techniques , Male , Mice , Mice, Inbred C57BL , Muscle, Smooth/cytology , Muscle, Smooth/enzymology , Muscle, Smooth/pathology , Prostate/cytology , Prostate/pathology , Prostatic Hyperplasia/enzymology , Prostatic Hyperplasia/pathology , Prostatic Hyperplasia/physiopathology , Signal Transduction
17.
Peptides ; 47: 71-6, 2013 Sep.
Article in English | MEDLINE | ID: mdl-23850991

ABSTRACT

It has been previously shown that octopus venoms contain novel tachykinin peptides that despite being isolated from an invertebrate, contain the motifs characteristic of vertebrate tachykinin peptides rather than being more like conventional invertebrate tachykinin peptides. Therefore, in this study we examined the effect of three variants of octopus venom tachykinin peptides on invertebrate and vertebrate tissues. While there were differential potencies between the three peptides, their relative effects were uniquely consistent between invertebrate and vertebrae tissue assays. The most potent form (OCT-TK-III) was not only the most anionically charged but also was the most structurally stable. These results not only reveal that the interaction of tachykinin peptides is more complex than previous structure-function theories envisioned, but also reinforce the fundamental premise that animal venoms are rich resources of novel bioactive molecules, which are useful investigational ligands and some of which may be useful as lead compounds for drug design and development.


Subject(s)
Ileum/drug effects , Mollusk Venoms/chemistry , Muscle Contraction/drug effects , Muscle, Smooth/drug effects , Tachykinins/pharmacology , Amino Acid Sequence , Animals , Astacoidea/drug effects , Astacoidea/physiology , Ileum/physiology , Male , Molecular Sequence Data , Muscle, Smooth/physiology , Octopodiformes/chemistry , Protein Binding , Rats , Receptors, Tachykinin/chemistry , Receptors, Tachykinin/metabolism , Sequence Alignment , Sequence Homology, Amino Acid , Tachykinins/chemical synthesis
18.
Organogenesis ; 9(3): 206-15, 2013.
Article in English | MEDLINE | ID: mdl-23872639

ABSTRACT

The adult prostate gland grows and develops under hormonal control while its physiological functions are controlled by the autonomic nervous system. The prostate gland receives sympathetic input via the hypogastric nerve and parasympathetic input via the pelvic nerve. In addition, the hypogastric and pelvic nerves also provide sensory inputs to the gland. This review provides a summary of the innervation of the adult prostate gland and describes the changes which occur with age and disease. Growth and development of the prostate gland is age dependent as is the occurrence of both benign prostate disease and prostate cancer. In parallel, the activity and influence of both the sympathetic and parasympathetic nervous system changes with age. The influence of the sympathetic nervous system on benign prostatic hyperplasia is well documented and this review considers the possibility of a link between changes in autonomic innervation and prostate cancer progression.


Subject(s)
Autonomic Nervous System , Prostate , Prostatic Neoplasms , Acetylcholine , Adult , Animals , Disease Progression , Humans , Male , Mice , Middle Aged , Norepinephrine , Prostate/innervation , Prostate/physiology , Prostatic Hyperplasia , Receptors, Adrenergic , Receptors, Muscarinic , Young Adult
19.
ChemMedChem ; 7(12): 2122-33, 2012 Dec.
Article in English | MEDLINE | ID: mdl-23055449

ABSTRACT

Noscapine is a phthalideisoquinoline alkaloid isolated from the opium poppy Papaver somniferum. It has long been used as an antitussive agent, but has more recently been found to possess microtubule-modulating properties and anticancer activity. Herein we report the synthesis and pharmacological evaluation of a series of 6'-substituted noscapine derivatives. To underpin this structure-activity study, an efficient synthesis of N-nornoscapine and its subsequent reduction to the cyclic ether derivative of N-nornoscapine was developed. Reaction of the latter with a range of alkyl halides, acid chlorides, isocyanates, thioisocyanates, and chloroformate reagents resulted in the formation of the corresponding N-alkyl, N-acyl, N-carbamoyl, N-thiocarbamoyl, and N-carbamate derivatives, respectively. The ability of these compounds to inhibit cell proliferation was assessed in cell-cycle cytotoxicity assays using prostate cancer (PC3), breast cancer (MCF-7), and colon cancer (Caco-2) cell lines. Compounds that showed activity in the cell-cycle assay were further evaluated in cell viability assays using PC3 and MCF-7 cells.


Subject(s)
Antineoplastic Agents, Phytogenic/chemistry , Antineoplastic Agents, Phytogenic/pharmacology , Noscapine/analogs & derivatives , Noscapine/pharmacology , Antineoplastic Agents, Phytogenic/chemical synthesis , Antitussive Agents/chemical synthesis , Antitussive Agents/chemistry , Antitussive Agents/pharmacology , Cell Cycle/drug effects , Cell Line, Tumor , Cell Proliferation/drug effects , Humans , Neoplasms/drug therapy , Noscapine/chemical synthesis , Papaver/chemistry
20.
J Biol Chem ; 287(16): 12952-65, 2012 Apr 13.
Article in English | MEDLINE | ID: mdl-22371491

ABSTRACT

We have provided the first evidence for specific heteromerization between the α(1A)-adrenoceptor (α(1A)AR) and CXC chemokine receptor 2 (CXCR2) in live cells. α(1A)AR and CXCR2 are both expressed in areas such as the stromal smooth muscle layer of the prostate. By utilizing the G protein-coupled receptor (GPCR) heteromer identification technology on the live cell-based bioluminescence resonance energy transfer (BRET) assay platform, our studies in human embryonic kidney 293 cells have identified norepinephrine-dependent ß-arrestin recruitment that was in turn dependent upon co-expression of α(1A)AR with CXCR2. These findings have been supported by co-localization observed using confocal microscopy. This norepinephrine-dependent ß-arrestin recruitment was inhibited not only by the α(1)AR antagonist Terazosin but also by the CXCR2-specific allosteric inverse agonist SB265610. Furthermore, Labetalol, which is marketed for hypertension as a nonselective ß-adrenoceptor antagonist with α(1)AR antagonist properties, was identified as a heteromer-specific-biased agonist exhibiting partial agonism for inositol phosphate production but essentially full agonism for ß-arrestin recruitment at the α(1A)AR-CXCR2 heteromer. Finally, bioluminescence resonance energy transfer studies with both receptors tagged suggest that α(1A)AR-CXCR2 heteromerization occurs constitutively and is not modulated by ligand. These findings support the concept of GPCR heteromer complexes exhibiting distinct pharmacology, thereby providing additional mechanisms through which GPCRs can potentially achieve their diverse biological functions. This has important implications for the use and future development of pharmaceuticals targeting these receptors.


Subject(s)
Prostate/metabolism , Protein Structure, Quaternary , Receptors, Adrenergic, alpha-1/chemistry , Receptors, Interleukin-8B/chemistry , Adrenergic alpha-1 Receptor Antagonists/pharmacology , Adrenergic alpha-Agonists/pharmacology , Allosteric Regulation/physiology , Animals , Arrestins/metabolism , CHO Cells , Chemokines/metabolism , Cricetinae , HEK293 Cells , Humans , Inositol Phosphates/metabolism , Labetalol/pharmacology , Male , Mice , Mice, Inbred C57BL , Norepinephrine/pharmacology , Prazosin/analogs & derivatives , Prazosin/pharmacology , Receptors, Adrenergic, alpha-1/metabolism , Receptors, G-Protein-Coupled/chemistry , Receptors, G-Protein-Coupled/metabolism , Receptors, Interleukin-8B/metabolism , beta-Arrestins
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