RESUMO
One cannot survive without regularly urinating and defecating. People with neurologic injury (spinal cord injury, traumatic brain injury, stroke) or disease (multiple sclerosis, Parkinson's disease, spina bifida) and many elderly are unable to voluntarily initiate voiding. The great majority of them require bladder catheters to void urine and "manual bowel programs" with digital rectal stimulation and manual extraction to void stool. Catheter-associated urinary tract infections frequently require hospitalization, whereas manual bowel programs are time consuming (1 to 2 hours) and stigmatizing and cause rectal pain and discomfort. Laxatives and enemas produce defecation, but onset and duration are unpredictable, prolonged, and difficult to control, which can produce involuntary defecation and fecal incontinence. Patients with spinal cord injury (SCI) consider recovery of bladder and bowel function a higher priority than recovery of walking. Bladder and bowel dysfunction are a top reason for institutionalization of elderly. Surveys indicate that convenience, rapid onset and short duration, reliability and predictability, and efficient voiding are priorities of SCI individuals. Despite the severe, unmet medical need, there is no literature regarding on-demand, rapid-onset, short-duration, drug-induced voiding therapies. This article provides in-depth discussion of recent discovery and development of two candidates for on-demand voiding therapies. The first, [Lys3,Gly8,-R-γ-lactam-Leu9]-NKA(3-10) (DTI-117), a neurokinin2 receptor agonist, induces both urination and defecation after systemic administration. The second, capsaicin (DTI-301), is a transient receptor potential cation channel subfamily V member 1 (TRPV1) receptor agonist that induces defecation after intrarectal administration. The review also presents clinical studies of a combination drug therapy administered via iontophoresis and preclinical studies of neuromodulation devices that induce urination and defecation. SIGNIFICANCE STATEMENT: A safe and effective, on-demand, rapid-onset, short-duration, drug-induced, voiding therapy could eliminate or reduce need for bladder catheters, manual bowel programs, and colostomies in patient populations that are unable to voluntarily initiate voiding. People with spinal injury place more importance on restoring bladder and bowel control than restoring their ability to walk. This paradigm-changing therapy would reduce stigmatism and healthcare costs while increasing convenience and quality of life.
Assuntos
Micção , Humanos , Micção/fisiologia , Micção/efeitos dos fármacos , Traumatismos da Medula Espinal/terapia , Traumatismos da Medula Espinal/fisiopatologia , Animais , Defecação/fisiologia , Defecação/efeitos dos fármacosRESUMO
Substantial clinical and preclinical evidence indicates that transient receptor potential vanilloid 1 (TRPV1) receptors are expressed on terminals of colorectal chemoreceptors and mechanoreceptors and are involved in various rectal hypersensitivity disorders with common features of colorectal overactivity. These stimulatory properties of TRPV1 receptors on colorectal function suggested that brief stimulation of TRPV1 might provide a means of pharmacologically activating the colorectum to induce defecation in patients with an "unresponsive" colorectum. The current studies explored the basic features of TRPV1 receptor-induced contractions of the colorectum in anesthetized rats with and without acute spinal cord injury (aSCI). Cumulative concentration-response curves to intrarectal (IR) capsaicin (CAP) solutions (0.003%-3.0%) were performed in anesthetized aSCI and spinal intact rats. CAP produced an "inverted U," cumulative concentration-response curve with a threshold for inducing colorectal contractions at 0.01% and a peak response at 0.1% and slight decreases in responses up to 3%. Decreases in responses with concentrations >0.1% are due to a rapid desensitization (i.e., ≤30 minutes) of TRPV1 receptors to each successive dose. Desensitization appeared fully recovered within 24 hours in spinal intact rats. Colorectal contractions were completely blocked by atropine, indicating a reflexogenic activation of parasympathetic neurons, and responses were completely unaffected by a neurokinin 2 receptor antagonist, indicating that release of neurokinin A from afferent terminals and subsequent direct contractions of the smooth muscle was not involved. IR administration of three other TRPV1 receptor agonists produced similar results as CAP. SIGNIFICANCE STATEMENT: Individuals with spinal cord injury often lose control of defecation. Time-consuming bowel programs using digital stimulation of the rectum are used to empty the bowel. This study shows that intrarectal administration of the transient receptor potential vanilloid 1 (TRPV1) receptor agonist, capsaicin, can induce rapid-onset, short-duration colorectal contractions capable of inducing defecation in spinal cord injured and intact rats. Therefore, TRPV1 agonists show promise as potential therapeutics to induce defecation in individuals with neurogenic bowel.
Assuntos
Capsaicina , Colo , Contração Muscular , Ratos Sprague-Dawley , Canais de Cátion TRPV , Animais , Masculino , Ratos , Canais de Cátion TRPV/agonistas , Canais de Cátion TRPV/metabolismo , Capsaicina/farmacologia , Colo/efeitos dos fármacos , Colo/metabolismo , Contração Muscular/efeitos dos fármacos , Traumatismos da Medula Espinal/fisiopatologia , Traumatismos da Medula Espinal/metabolismo , Reto/efeitos dos fármacos , Reto/inervação , Relação Dose-Resposta a Droga , Anestesia , Fatores de TempoRESUMO
The feasibility of eliciting defecation and urination after intranasal (IN) or sublingual (SL) delivery of a small peptide NK2 receptor agonist, [Lys5, MeLeu9, Nle10]-NKA(4-10), was examined using prototype formulations in dogs. In anesthetized animals, administration of 100 or 300 µg/kg IN or 2.0-6.7 mg/kg SL increased colorectal peak pressure and area under the curve. Peak bladder pressure was also increased at the same doses, and this was accompanied by highly efficient voiding at normal physiological bladder pressure. The onset of these effects was rapid (≤2.5 min), and the primary contractions lasted â¼25 min, returning to baseline in <60 min. Slight hypotension lasting a few minutes and causing <10% change from baseline was detected after higher doses and was statistically significant after only 100 µg/kg IN. In conscious dogs, there was a dose-related increase in voiding responses and reduction in the latency to urinate and defecate after 300 and 1000 µg/kg IN; emesis was also observed at these doses. SL administration of 6.7 mg/kg induced urination within 10 min, but not defecation or emesis. These findings support the feasibility of developing a convenient dosage form of small peptide NK2 receptor agonists as on-demand defecation or urination therapies.
Assuntos
Neoplasias Colorretais , Bexiga Urinária , Cães , Animais , Receptores da Neurocinina-2/agonistas , Neurocinina A/farmacologia , Peptídeos/farmacologia , VômitoRESUMO
STUDY DESIGN: Preclinical pharmacology. OBJECTIVES: To determine whether blocking substance P signaling attenuates the hypertension and bradycardia evoked by colorectal distension (CRD) in spinal cord injured (SCI) rats. SETTING: University laboratory in Pennsylvania, U.S.A. METHODS: Tachykinin NK1 receptor antagonists were administered 30 min prior to CRD three weeks after complete spinal cord transection at the 4th thoracic (T4) level. The dose range, route of administration, and pretreatment time was based on published data demonstrating occupancy of brain NK1 receptors in rodents. RESULTS: Subcutaneous (SC) administration of 10-30 mg/kg GR205171 ((2S,3S)-N-[[2-methoxy-5-[5-(trifluoromethyl)tetrazol-1-yl]phenyl]methyl]-2-phenylpiperidin-3-amine dihydrochloride) reduced CRD-induced hypertension and bradycardia by 55 and 49%, respectively, compared with pretreatment values. There was no effect of GR205171 on resting blood pressure or heart rate. In contrast, the same dose range of CP-99,994 ((2S,3S)-N-[(2-methoxyphenyl)methyl]-2-phenyl-3-piperidinamine dihydrochloride) had no effect on CRD-induced cardiovascular responses. CONCLUSIONS: The effective dose range of GR205171 to alleviate autonomic dysreflexia is consistent with the blockade of NK1 receptors on pelvic sensory afferents in the lumbosacral spinal cord, which may in turn prevent the over-excitation of sympathetic preganglionic neurons (SPNs) that regulate blood pressure and heart rate. The findings provide preclinical support for the utility of NK1 receptor antagonists to treat autonomic dysreflexia in people with SCI. The difference in the effects of the two NK1 receptor antagonists may reflect the ~200-fold lower affinity of CP-99,994 than GR205171 for the rat NK1 receptor.
Assuntos
Disreflexia Autonômica , Neoplasias Colorretais , Hipertensão , Traumatismos da Medula Espinal , Ratos , Animais , Disreflexia Autonômica/tratamento farmacológico , Disreflexia Autonômica/etiologia , Traumatismos da Medula Espinal/complicações , Traumatismos da Medula Espinal/tratamento farmacológico , Bradicardia/tratamento farmacológico , Bradicardia/etiologia , Ratos Wistar , Medula EspinalRESUMO
STUDY DESIGN: Animal proof of principle study. OBJECTIVES: Bladder and bowel dysfunction are common after spinal cord injury (SCI) and in the elderly. Neurokinin 2 receptor agonists are known to produce on-demand urination and defecation in adult SCI rats. This study compared the ability of a neurokinin 2 receptor (NK2R) agonist to produce bladder and colorectal contractions in both young adult and aged SCI rats. SETTING: Dignify Therapeutics and Integrated Laboratory Systems, Durham, NC USA. METHODS: Bladder and colorectal pressure and voiding efficiency were measured in response to the NK2R agonist, [Lys5,Me,Leu9,Nle10]-NKA(4-10) (LMN-NKA), in anesthetized animals. The potency and efficacy of LMN-NKA was examined in young adult and aged SCI (T3 or T9 transected) rats, with young adult and aged spinal intact rats included as controls. RESULTS: LMN-NKA (3-300 µg/kg i.v.) produced dose-dependent increases in bladder and colorectal pressure in all anesthetized rats. No differences in the bladder or colorectal pressure responses or voiding efficiency were observed with age or after SCI. The level of SCI did not change the pharmacodynamic responses to the agonist. CONCLUSIONS: An NK2R agonist produced similar responses in young adult and aged SCI rats, suggesting this class of agonists could be used as a potential therapy to induce on-demand urination and defecation in aged populations, with or without SCI.
Assuntos
Neoplasias Colorretais , Traumatismos da Medula Espinal , Animais , Ratos , Ratos Sprague-Dawley , Receptores da Neurocinina-2 , Medula Espinal , Traumatismos da Medula Espinal/complicações , Traumatismos da Medula Espinal/tratamento farmacológico , Bexiga Urinária , MicçãoRESUMO
STUDY DESIGN: Survey. OBJECTIVES: To investigate the needs and priorities of people with spinal cord injury for managing neurogenic bladder and bowel function and to determine their willingness to adopt neuromodulation interventions for these functions. METHODS: Anonymous online survey. It was advertised by word-of-mouth by community influencers and social media, and by advertisement in newsletters of advocacy groups. RESULTS: Responses from 370 individuals (27% female, 73% male) were included. Bladder emptying without catheters was the top priority for restoring bladder function, and maintaining fecal continence was the top priority for restoring bowel function. The biggest concerns regarding external stimulation systems were wearing a device with wires connecting to electrodes on the skin and having to don and doff the system daily as needed. The biggest concerns for implanted systems were the chances of experiencing problems with the implant that required a revision surgery or surgical removal of the whole system. Respondents were willing to accept an external (61%) or implanted (41%) device to achieve improved bladder or bowel function. CONCLUSIONS: Bladder and bowel dysfunction remain important unmet challenges for individuals living with SCI who answered our survey. These individuals are willing to accept some potential risks of nerve stimulation approaches given potential benefits. Additional consumer input is critical for guiding both research and translation to clinical use and personalized medicine.
Assuntos
Traumatismos da Medula Espinal , Bexiga Urinaria Neurogênica , Atitude , Feminino , Humanos , Masculino , Traumatismos da Medula Espinal/complicações , Traumatismos da Medula Espinal/terapia , Inquéritos e Questionários , Bexiga Urinaria Neurogênica/etiologia , Bexiga Urinaria Neurogênica/terapiaRESUMO
Tachykinin neurokinin 2 (NK2) receptor agonists may have potential to alleviate clinical conditions associated with bladder and gastrointestinal underactivity by stimulating contraction of visceral smooth muscle. The ability of [Lys5,MeLeu9,Nle10]-neurokinin A(4-10) (LMN-NKA) to elicit micturition and defecation was examined after repeated administration in groups of 2-10 conscious dogs. Administration of 10-100 µg/kg, i.v., four times daily for six consecutive days, reliably elicited micturition after ≥90% of doses and defecation after ≥50% of doses. Voiding occurred <4 minutes after dosing and was short lasting (<10 minutes). LMN-NKA was well tolerated, with emesis after â¼25% of doses at 100 µg/kg, i.v. Hypotension was induced by 100 µg/kg, i.v., of LMN-NKA but not by lower doses. Administration of 30-300 µg/kg, s.c., twice daily for seven consecutive days, reliably elicited both urination and defecation after 88%-100% of doses, and was accompanied by a high rate of emesis (50%-100%). The onset of voiding was rapid (<7 minutes) but was more prolonged than after intravenous administration (30-60 minutes). Emesis induced by 30 or 300 µg/kg, s.c., of LMN-NKA was significantly reduced (from 58% to 8% and from 96% to 54%, respectively) by a 30-minute pretreatment with the neurokinin 1 (NK1) receptor antagonist, (2S,3S)-N-(2-methoxybenzyl)-2-phenylpiperidin-3-amine (CP-99,994; 1 mg/kg, s.c.). The ability of selective NK2 receptor agonists to elicit on-demand voiding could potentially address a major unmet need in people lacking voluntary control of micturition and/or defecation. LMN-NKA unexpectedly activated NK1 receptors at doses that stimulated voiding, causing emesis and hypotension that may limit the clinical utility of nonselective NK2 receptor agonists.
Assuntos
Defecação/efeitos dos fármacos , Hipotensão/induzido quimicamente , Fragmentos de Peptídeos/farmacologia , Receptores da Neurocinina-1/metabolismo , Receptores da Neurocinina-2/metabolismo , Micção/efeitos dos fármacos , Vômito/induzido quimicamente , Animais , Estado de Consciência , Cães , Neurocinina A/química , Fragmentos de Peptídeos/efeitos adversos , Fragmentos de Peptídeos/farmacocinética , Distribuição TecidualRESUMO
Ejaculation is controlled by coordinated and rhythmic contractions of bulbospongiosus (BSM) and ischiocavernosus muscles. Motoneurons that innervate and control BSM contractions are located in the dorsomedial portion of the ventral horn in the L(5-6) spinal cord termed the dorsomedial (DM) nucleus. We characterized intrinsic properties of DM motoneurons as well as synaptic inputs from the dorsal gray commissure (DGC). Electrical stimulation of DGC fibers elicited fast inhibitory and excitatory responses. In the presence of glutamate receptor antagonists, both fast GABAergic as well as glycinergic inhibitory postsynaptic potentials (IPSPs) were recorded. No slow GABA(B)-mediated inhibition was evident. In the presence of GABA(A) and glycine receptor antagonists, DGC stimulation elicited fast glutamatergic excitatory responses that were blocked by application of CNQX. Importantly, a slow depolarization (timescale of seconds) was routinely observed that sufficiently depolarized the DM motoneurons to fire "bursts" of action potentials. This slow depolarization was elicited by a range of stimulus train frequencies and was insensitive to glutamate receptor antagonists (CNQX and d-APV). The slow depolarization was accompanied by an increase in membrane resistance with an extrapolated reversal potential near the K(+) Nernst potential. It was mediated by the combination of the block of a depolarization-activated K(+) current and the activation of a QX-314-sensitive cation current. These results demonstrate that fast synaptic responses in DM motoneurons are mediated primarily by glutamate, GABA, and glycine receptors. In addition, slow nonglutamatergic excitatory postsynaptic potentials (EPSPs), generated through DGC stimulation, can elicit burstlike responses in these neurons.
Assuntos
Ejaculação/fisiologia , Neurônios Motores/fisiologia , Músculo Esquelético/inervação , Medula Espinal/fisiologia , Sinapses/fisiologia , 2-Amino-5-fosfonovalerato/farmacologia , 6-Ciano-7-nitroquinoxalina-2,3-diona/farmacologia , Potenciais de Ação/efeitos dos fármacos , Animais , Estimulação Elétrica , Antagonistas de Aminoácidos Excitatórios/farmacologia , Potenciais Pós-Sinápticos Excitadores/efeitos dos fármacos , Potenciais Pós-Sinápticos Excitadores/fisiologia , Potenciais Pós-Sinápticos Inibidores/efeitos dos fármacos , Potenciais Pós-Sinápticos Inibidores/fisiologia , Masculino , Neurônios Motores/efeitos dos fármacos , Músculo Esquelético/fisiologia , Técnicas de Patch-Clamp , Ratos , Ratos Sprague-Dawley , Medula Espinal/efeitos dos fármacos , Sinapses/efeitos dos fármacos , Transmissão Sináptica/efeitos dos fármacos , Transmissão Sináptica/fisiologiaRESUMO
The urethrogenital reflex (UGR) is used as a surrogate model of the autonomic and somatic nerve and muscle activity that accompanies ejaculation. The UGR is evoked by distension of the urethra and activation of penile afferents. The current study compares two methods of elevating urethral intraluminal pressure in spinalized, anesthetized male Sprague-Dawley rats (n = 60). The first method, penile extension UGR, involves extracting the penis from the foreskin, so that urethral pressure rises due to a natural anatomical flexure in the penis. The second method, penile clamping UGR, involves penile extension UGR with the addition of clamping of the glans penis. Groups of animals were prepared that either received no additional treatment, surgical shams, or received bilateral nerve cuts (4 nerve cut groups): either the pudendal sensory nerve branch (SbPN), the pelvic nerves, the hypogastric nerves, or all three nerves. Penile clamping UGR was characterized by multiple bursts, monitored by electromyography (EMG) of the bulbospongiosus muscle (BSM) accompanied by elevations in urethral pressure. The penile clamping UGR activity declined across multiple trials and eventually resulted in only a single BSM burst, indicating desensitization. In contrast, the penile extension UGR, without penile clamping, evoked only a single BSM EMG burst that showed no desensitization. Thus, the UGR is composed of two BSM patterns: an initial single burst, termed urethrobulbospongiosus (UBS) reflex and a subsequent multiple bursting pattern (termed ejaculation-like response, ELR) that was only induced with penile clamping urethral occlusion. Transection of the SbPN eliminated the ELR in the penile clamping model, but the single UBS reflex remained in both the clamping and extension models. Pelvic nerve (PelN) transection increased the threshold for inducing BSM activation with both methods of occlusion but actually unmasked an ELR in the penile extension method. Hypogastric nerve (HgN) cuts did not significantly alter any parameter. Transection of all three nerves eliminated BSM activation completely. In conclusion, penile clamping occlusion recruits penile and urethral primary afferent fibers that are necessary for an ELR. Urethral distension without significant penile afferent activation recruits urethral primary afferent fibers carried in either the pelvic or pudendal nerve that are necessary for the single-burst UBS reflex.
Assuntos
Glândulas Bulbouretrais/fisiologia , Contração Muscular/fisiologia , Músculo Liso/fisiologia , Uretra/fisiopatologia , Animais , Ejaculação/fisiologia , Estimulação Elétrica , Eletromiografia , Masculino , Modelos Animais , Neurônios Aferentes/fisiologia , Pênis/fisiologia , Ratos , Ratos Sprague-DawleyRESUMO
The urethral rhabdosphincter (URS), commonly known as the external urethral sphincter, facilitates urinary continence by constricting the urethra. Striated muscle fibres in the urethral rhabdosphincter are innervated by Onuf's nuclei motoneurons in the spinal cord. Although noradrenaline (NA) reuptake inhibitors are shown to increase URS tone preventing urinary leakage in incontinent patients, whether or how NA affects URS motoneurons is unknown. Properties of dye-labelled URS motoneurons were investigated by whole-cell patch-clamp recordings in isolated spinal cord slices prepared from neonatal female rats. As previously shown for adult sphincter motoneurons, neonatal URS motoneurons are more depolarized and possess higher input resistance than other spinal α-motoneurons. These distinct properties make URS motoneurons more excitable than other α-motoneurons. Moreover, bath application of noradrenaline (NA) significantly depolarizes URS motoneurons and in many cases evokes action potentials. NA also significantly increases input resistance and reduces rheobase. These changes are reversed with wash, are largely blocked by the α(1)-adrenoceptor-selective antagonist prazosin, and are mimicked by the α(1)-adrenoceptor-selective agonist phenylephrine. In addition, NA significantly reduces the amplitude of the afterhyperpolarization and increases action potential frequency. Both the increase in action potential frequency and the reduction in afterhyperpolarization are occluded by apamin, a small-conductance calcium-activated potassium (SK(Ca)) channel blocker. In conclusion, NA effectively increases the excitability of URS motoneurons through multiple mechanisms. The NA-induced increase in excitability of urethral rhabdosphincter motoneurons could be a key mechanism by which NA reuptake inhibitors improve stress urinary incontinence.
Assuntos
Neurônios Motores/fisiologia , Uretra/fisiologia , Animais , Animais Recém-Nascidos , Feminino , Potenciais da Membrana/fisiologia , Norepinefrina/fisiologia , Técnicas de Patch-Clamp , Canais de Potássio Cálcio-Ativados/metabolismo , Ratos , Ratos Endogâmicos , Receptores Adrenérgicos alfa 1/fisiologiaRESUMO
The urethral rhabdosphincter and pelvic floor muscles are important in maintenance of urinary continence and in preventing descent of pelvic organs [i.e., pelvic organ prolapse (POP)]. Despite its clinical importance and complexity, a comprehensive review of neural control of the rhabdosphincter and pelvic floor muscles is lacking. The present review places historical and recent basic science findings on neural control into the context of functional anatomy of the pelvic muscles and their coordination with visceral function and correlates basic science findings with clinical findings when possible. This review briefly describes the striated muscles of the pelvis and then provides details on the peripheral innervation and, in particular, the contributions of the pudendal and levator ani nerves to the function of the various pelvic muscles. The locations and unique phenotypic characteristics of rhabdosphincter motor neurons located in Onuf's nucleus, and levator ani motor neurons located diffusely in the sacral ventral horn, are provided along with the locations and phenotypes of primary afferent neurons that convey sensory information from these muscles. Spinal and supraspinal pathways mediating excitatory and inhibitory inputs to the motor neurons are described; the relative contributions of the nerves to urethral function and their involvement in POP and incontinence are discussed. Finally, a detailed summary of the neurochemical anatomy of Onuf's nucleus and the pharmacological control of the rhabdosphincter are provided.
Assuntos
Canal Anal/inervação , Defecação , Neurônios Motores/fisiologia , Neurônios Aferentes/fisiologia , Diafragma da Pelve/inervação , Uretra/inervação , Micção , Animais , Defecação/efeitos dos fármacos , Feminino , Humanos , Neurônios Motores/efeitos dos fármacos , Inibição Neural , Neurônios Aferentes/efeitos dos fármacos , Parto , Prolapso de Órgão Pélvico/fisiopatologia , Fenótipo , Reflexo , Sensação , Incontinência Urinária/fisiopatologia , Micção/efeitos dos fármacosRESUMO
Urine storage is facilitated by somatic (pudendal nerve) and sympathetic [hypogastric nerve (HgN)] reflexes to the urethral rhabdosphincter (URS) and urethral smooth muscle, respectively, initiated by primary afferent fibers in the pelvic nerve (PelN). Inhibition of storage reflexes is required for normal voiding. This study characterizes a urine storage reflex inhibitory network that can be activated by PelN afferent fibers concurrently with the reflexes themselves. Electrical stimulation of PelN produced evoked potentials recorded by URS EMG electrodes (10-ms latency) or HgN electrodes (60-ms latency) in chloralose-anesthetized cats. When a second (i.e., paired) pulse of the same stimulus intensity was applied to the PelN 50-500 ms after the first, the reflexes evoked by the second stimulus were inhibited. The inhibition was maximal at paired-pulse intervals of 50-100 ms and remained after acute spinal transection at T10, confirming that the inhibitory center is located in the spinal cord. The 5-HT(1A) receptor agonist 8-hydroxy-2-(di-n-propylamino)tertralin (8-OH-DPAT; 3-300 mug/kg iv) consistently reduced the paired-pulse inhibition from 20% to 60% of control in spinal-intact animals but had no effect in acute spinal animals (i.e., supraspinal site of action). N-{2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl}-N-2-pyridinylcyclohexanecarboxamide maleate (300 mug/kg iv) completely reversed 8-OH-DPAT's effects. The PelN-HgN reflex paired-pulse inhibition was not affected by 8-OH-DPAT. These results indicate the presence of a spinal, urine storage reflex, inhibitory center (SUSRIC) that is activated within 50 ms after activation of the reflexes themselves. SUSRIC is inhibited (disfacilitated) by supraspinal 5-HT(1A) receptors.
Assuntos
Inibição Neural/fisiologia , Receptor 5-HT1A de Serotonina/fisiologia , Medula Espinal/fisiologia , Bexiga Urinária/inervação , Bexiga Urinária/fisiologia , Micção/fisiologia , 8-Hidroxi-2-(di-n-propilamino)tetralina/farmacologia , Animais , Gatos , Estimulação Elétrica , Eletromiografia , Potenciais Evocados/efeitos dos fármacos , Potenciais Evocados/fisiologia , Feminino , Interneurônios/efeitos dos fármacos , Interneurônios/fisiologia , Neurônios Motores/efeitos dos fármacos , Neurônios Motores/fisiologia , Inibição Neural/efeitos dos fármacos , Piperazinas/farmacologia , Piridinas/farmacologia , Reflexo/fisiologia , Antagonistas da Serotonina/farmacologia , Agonistas do Receptor de Serotonina/farmacologia , Medula Espinal/efeitos dos fármacos , Uretra/inervação , Uretra/fisiologiaRESUMO
Acute administration of [Lys5,Me,Leu9,Nle10]-NKA(4-10) (LMN-NKA) produces contractions of the detrusor and rectum with voiding in intact and acutely spinal cord injured (SCI) rats. In the current study, the ability of LMN-NKA (10 µg/kg or 100 µg/kg, subcutaneous [SC], twice a day [bid]) or vehicle to induce voiding and defecation in chronic SCI rats was examined across 30 days. After the last day of administration, voiding response rates and bladder pressure (BP) responses to LMN-NKA (intravenous [IV] and SC) were evaluated under anesthesia. In conscious rats, LMN-NKA (100 µg/kg) produced dose-dependent micturition within 5 min, with response rates >90%, and voiding efficiency >80% in males and >60% in females, which remained stable across the 1-month test period. Similarly, LMN-NKA administration rapidly induced defecation, which also remained stable. Under anesthesia, LMN-NKA increased BP, voiding efficiency, and voiding response rates, which reached 100% at 3 and 10 µg/kg IV in males and females, respectively. SC administration produced 100% response rates in males (30 µg/kg) but only 71% in females (100 µg/kg). Efficacy in rats chronically treated with LMN-NKA was similar to naïve and vehicle-treated rats, except for reduced voiding efficiency in chronically dosed female rats (100 µg/kg). No differences in bladder weights or collagen-to-smooth muscle ratios in histological sections were seen between the groups. Thus neither tolerance, nor sensitization, to LMN-NKA-induced micturition and defecation occurs with chronic administration in rats with chronic SCI. Efficacy was higher in male than in female rats.
Assuntos
Defecação/efeitos dos fármacos , Fragmentos de Peptídeos/administração & dosagem , Receptores da Neurocinina-2/agonistas , Traumatismos da Medula Espinal/tratamento farmacológico , Micção/efeitos dos fármacos , Animais , Defecação/fisiologia , Relação Dose-Resposta a Droga , Esquema de Medicação , Feminino , Masculino , Ratos , Ratos Sprague-Dawley , Receptores da Neurocinina-2/metabolismo , Traumatismos da Medula Espinal/metabolismo , Traumatismos da Medula Espinal/fisiopatologia , Vértebras Torácicas/lesões , Micção/fisiologiaRESUMO
The affinity, potency, efficacy, and selectivity of the NK2 receptor agonist GR64349 ([Lys3,Gly8,-R-γ-lactam-Leu9]NKA(3-10)) at human recombinant NK2 and NK1 receptors was examined. In radioligand binding studies, GR64349 displaced [125I]-NKA binding to NK2 receptors with high affinity (pKi 7.77â¯+â¯0.10) but only weakly displaced [3H]-septide binding to NK1 receptors (pKi <5). In functional studies examining increases in intracellular inositol-1 phosphate (IP-1) accumulation, calcium levels, and cyclic AMP synthesis, GR64349 was a full agonist by reference to the endogenous agonists NKA (NK2 receptors) and substance P (NK1 receptors). GR64349 increased IP-1 accumulation with 1,400-fold greater potency in cells expressing NK2 receptors (pEC50 9.10â¯+â¯0.16) than cells expressing NK1 receptors (pEC50 5.95â¯+â¯0.80). For calcium responses, GR64349 was 500-fold more potent in the assay using NK2 receptors (pEC50 9.27 + 0.26) than NK1 receptors (pEC50 6.55â¯+â¯0.16). GR64349 also stimulated cyclic AMP synthesis in both cell lines, and was almost 900-fold more potent at NK2 receptors (pEC50 10.66â¯+â¯0.27) than NK1 receptors (pEC50 7.71â¯+â¯0.41). These findings confirm that GR64349 is the most selective NK2 receptor agonist described to date.
Assuntos
Receptores da Neurocinina-1/agonistas , Receptores da Neurocinina-2/agonistas , Animais , Células CHO , Cricetulus , Humanos , Proteínas Recombinantes/efeitos dos fármacosRESUMO
The effects of the neurokinin NK2 receptor agonist [Lys5,MeLeu9,Nle10]-NKA(4-10) (LMN-NKA) on bladder and colorectal function were examined in minipigs. In anesthetized animals, subcutaneous (SC) administration of 30-100⯵g/kg increased peak bladder and colorectal pressures. Increases in bladder and colorectal pressure were inhibited by a 15â¯min pretreatment with the NK2 receptor antagonist GR 159897 (1â¯mg/kg intravenously (IV)). Bladder and colorectal pressures were also increased after IV (0.3⯵g/kg), intranasal (IN; 100⯵g/kg) and sublingual administration (SL; 5â¯mg/kg). There was a nonsignificant trend for hypotension (16 or 12% decrease in mean arterial pressure) after 100⯵g/kg SC and 0.3⯵g/kg IV, respectively, but not after 100⯵g/kg IN or 5â¯mg/kg SL. In conscious minipigs, 30-300⯵g/kg SC caused a dose-related increase in defecation that was accompanied by emesis in 38% of subjects receiving 300⯵g/kg. Urination was increased after 100⯵g/kg SC but not lower or higher doses. The peak plasma exposure (Cmax) after 100⯵g/kg SC was 123â¯ng/mL, and area under the curve (AUC) was 1790â¯minâ¯*â¯ng/mL. Defecation response rates (~82%) were maintained after SC administration of LMN-NKA (30⯵g/kg) given 3 times daily over 5 consecutive days. Defecation rates were higher after a single dose of 100⯵g/kg IN compared with vehicle, but this did not reach significance. After 7-10â¯mg/kg SL, 83% of animals urinated and defecated, and none had emesis. The data support the feasibility of developing a convenient and well-tolerated route of administration of LMN-NKA for human use. Minipigs may be a suitable species for toxicology studies with LMN-NKA due to the relatively low rate of emesis in this species.
Assuntos
Colo/efeitos dos fármacos , Defecação/efeitos dos fármacos , Receptores da Neurocinina-2/agonistas , Reto/efeitos dos fármacos , Bexiga Urinária/efeitos dos fármacos , Micção/efeitos dos fármacos , Animais , Indóis/farmacologia , Piperidinas/farmacologia , Pressão , Receptores da Neurocinina-2/antagonistas & inibidores , Suínos , Porco MiniaturaRESUMO
OBJECTIVE: To determine whether experimental denervation of the levator ani (LA) and its subsequent atrophy contribute to the development of pelvic organ prolapse in the squirrel monkey. STUDY DESIGN: Thirty-seven female monkeys were evaluated including 7 that underwent bilateral LA neurectomy (bLAN), 17 nulliparous monkeys without prolapse, 7 parous monkeys without prolapse, and 6 parous monkeys with prolapse. Magnetic resonance imaging was used to calculate LA muscle volumes and obtain measurements of the position of bladder and cervix. Repeat observations in bLAN females occurred at different times in relation to parturition. RESULTS: LA volumes were reduced in bLAN monkeys (P = .02). Bladder (P = .03) and cervix (P = .04) positions varied between groups, with nulliparous females having the most cephalad positions and females with prolapse having the most caudal positions. Bladder descent was observed in a subset of 4 bLAN females that experienced vaginal parturition (P = .04) and correlated with external findings of vaginal prolapse. CONCLUSION: Bilateral transection of the LA nerve results in atrophy of denervated LA muscles but not a loss of pelvic support in nulliparous monkeys, suggesting that connective tissue components compensate for weakened pelvic floor muscles. LA denervation may accelerate the onset of vaginal prolapse subsequent to parturition.
Assuntos
Músculo Liso/inervação , Prolapso Uterino/patologia , Prolapso Uterino/fisiopatologia , Animais , Atrofia , Tecido Conjuntivo/patologia , Feminino , Imageamento por Ressonância Magnética , Denervação Muscular , Parto , Pressão , Saimiri , UrodinâmicaRESUMO
The effects of the tachykinin NK2 receptor agonist LMN-NKA ([Lys5,MeLeu9,Nle10]-NKA(4-10)) on colorectal and arterial blood pressure were examined in anesthetized macaques. Intravenous (IV) administration of 1-100 µg/kg caused dose-related increases in colorectal pressure up to 120 mmHg above baseline, and area under the curve (AUC) up to 24,987 mmHg*s. This was accompanied at all doses by transient hypotension, with up to 26% reduction in mean arterial pressure (MAP) from baseline. Hypotension, but not the increase in colorectal pressure, was inhibited by a 10-min pretreatment with the NK1 receptor antagonist CP-99,994. In a pilot experiment using subcutaneous (SC) injection, a similar dose range of LMN-NKA (3-100 µg/kg) again appeared to increase colorectal pressure with a similar AUC (up to 18,546 mmHg*s) to that seen after IV injection, but lower peak amplitude (up to 49 mmHg). Unlike the effects of IV injection, hypotension was only present after the highest SC dose (100 µg/kg) in one of two animals. Pharmacokinetic analysis revealed markedly lower plasma exposures after SC compared with IV administration. Cmax was 39.6 versus 1070 ng/mL, and AUCinf was 627 versus 2090 ng/mL*min, respectively. These findings are consistent with previous observations in anesthetized dogs and indicate that the prokinetic effects of LMN-NKA may be achieved without hypotension using a route of administration that avoids unnecessarily high plasma exposures.
Assuntos
Pressão Arterial/efeitos dos fármacos , Colo/efeitos dos fármacos , Neurocinina A/análogos & derivados , Neurocinina A/administração & dosagem , Receptores da Neurocinina-2/agonistas , Reto/efeitos dos fármacos , Administração Intravenosa , Anestesia , Animais , Colo/fisiologia , Feminino , Injeções Subcutâneas , Macaca , Masculino , Neurocinina A/sangue , Reto/fisiologiaRESUMO
Tachykinin NK2 receptor (NK2R) agonists have potential to alleviate clinical conditions associated with bladder and gastrointestinal under activity. The effects of agonists with differing selectivity for NK2R over NK1Rs on colorectal, bladder, and cardiovascular function were examined in anesthetized dogs. Intravenous (IV) administration of NKA, LMN-NKA ([Lys5,MeLeu9,Nle10]-NKA(4-10)), and [ß-Ala8]-NKA(4-10) caused a dose-related increase in colorectal pressure (up to 98 mmHg) that was blocked by pretreatment with the NK2R antagonist GR 159897 (1 mg/kg), and hypotension (decrease in mean arterial pressure of ~40 mmHg) that was blocked by the NK1R antagonist CP-99,994 (1 mg/kg). Despite the greater in vitro selectivity of LMN-NKA and [ß-Ala8]-NKA(4-10) for NK2R over NK1Rs compared with NKA, all 3 agonists increased colorectal pressure and caused hypotension within a similar dose range when administered as a bolus (0.1-300 µg/kg IV), or even as a slow IV infusion over 5 min (NKA; 0.02-0.6 µg/kg/min). In contrast, subcutaneous (SC) administration of LMN-NKA (3-10 µg/kg) increased colorectal pressure (up to 50 mmHg) and elicited micturition (⧠85% voiding efficiency) without causing hypotension. NK2R agonists can produce rapid-onset, short-duration, colorectal contractions, and efficient voiding of urine without hypotension after SC administration, indicating that routes of administration that avoid the high plasma concentrations associated with IV dosing improve the separation between desired and unwanted pharmacodynamic effects. The potent hypotensive effect of NKA in dogs was unexpected based on published studies in humans in which IV infusion of NKA did not affect blood pressure at doses that increased gastrointestinal motility.
Assuntos
Pressão Arterial/efeitos dos fármacos , Colo/efeitos dos fármacos , Neurocinina A/análogos & derivados , Neurocinina A/farmacologia , Receptores da Neurocinina-1/fisiologia , Receptores da Neurocinina-2/fisiologia , Bexiga Urinária/efeitos dos fármacos , Anestesia , Animais , Colo/fisiologia , Cães , Feminino , Frequência Cardíaca/efeitos dos fármacos , Indóis/farmacologia , Masculino , Antagonistas dos Receptores de Neurocinina-1/farmacologia , Piperidinas/farmacologia , Receptores da Neurocinina-2/agonistas , Receptores da Neurocinina-2/antagonistas & inibidores , Bexiga Urinária/fisiologiaRESUMO
The suitability of various neurokinin-2 (NK2) receptor agonists and routes of administration to elicit on-demand voiding of the bladder and bowel, as future therapy for individuals with spinal cord injury, was examined using a rat model. The current study examined the feasibility of alternative routes of administration, which are more practical for clinical use than intravenous (IV) administration. Voiding and isovolumetric cystometry were recorded in anesthetized, acutely spinalized, female rats after IV, subcutaneous (SC), intramuscular (IM), intranasal (IN), or sublingual (SL) administration of [Lys5,MeLeu9,Nle10]-NKA(4-10) (LMN-NKA). Administration of LMN-NKA (1-10µg/kg IV; 10-300µg/kg SC or IM; 15-1000µg/kg IN or 300-1500µg/kg SL) elicited rapid-onset, short-duration, dose-related increases in bladder pressure and voiding with the rank order for time of both onset and duration being IV < IN < SC = IM < SL. The incidence of voiding was dependent on the dose and route, with all routes resulting in a high voiding efficiency (~ 70%). Like LMN-NKA, neurokinin A (NKA 1-100µg/kg IV) and GR 64349 (0.1-30µg/kg IV or 1-300µg/kg SC) produced rapid-onset, short-duration increases in bladder pressure, as well as colorectal pressure. Administration of vehicle never produced bladder or rectal contractions or voiding. Transient hypotension was observed after IV injection of LMN-NKA, which was less pronounced after SC injection. Hypotension was not apparent with GR 64349. In conclusion, selective NK2 receptor agonists, administered through various non-IV routes of administration, may provide a safe, convenient, and efficacious method for inducing voiding.
Assuntos
Peptídeos/farmacologia , Receptores da Neurocinina-2/agonistas , Reto/efeitos dos fármacos , Reto/fisiopatologia , Traumatismos da Medula Espinal/fisiopatologia , Bexiga Urinária/efeitos dos fármacos , Bexiga Urinária/fisiopatologia , Animais , Relação Dose-Resposta a Droga , Feminino , Motilidade Gastrointestinal/efeitos dos fármacos , Contração Muscular/efeitos dos fármacos , Peptídeos/química , Peptídeos/farmacocinética , Pressão , Ratos , Ratos Sprague-DawleyRESUMO
A series of peptide NK2 receptor agonists was evaluated for affinity, potency, efficacy, and selectivity at human recombinant NK2 and NK1 receptors expressed in CHO cells to identify compounds with the greatest separation between NK2 and NK1 receptor agonist activity. Binding studies were performed using displacement of [125I]-NKA binding to NK2 receptors and displacement of [3H]-Septide binding to NK1 receptors expressed in CHO cells. Functional studies examining the increase in intracellular calcium levels and cyclic AMP stimulation were performed using the same cell lines. A correlation was demonstrated between binding affinities (Ki) and potency to increase intracellular calcium (EC50) for NK2 and NK1 receptors. Ranking compounds by their relative affinity (Ki) or potency (EC50) at NK2 or NK1 receptors indicated that the most selective NK2 agonists tested were [Lys5,MeLeu9,Nle10]-NKA(4-10) (NK1/NK2 Ki ratio = 674; NK1/NK2 EC50 ratio = 105) and [Arg5,MeLeu9,Nle10]-NKA(4-10) (NK1/NK2 Ki ratio = 561; NK1/NK2 EC50 ratio = 70). The endogenous peptide, NKA, lacked selectivity with an NK1/NK2 Ki ratio = 20 and NK1/NK2 EC50 ratio = 1. Of the compounds selected for evaluation in cyclic AMP stimulation assays, [ß-Ala8]-NKA(4-10) had the greatest selectivity for activation of NK2 over NK1 receptors (NK1/NK2 EC50 ratio = 244), followed by [Lys5,MeLeu9,Nle10]-NKA(4-10) (ratio = 74), and NKA exhibited marginal selectivity (ratio = 2.8).