Interaction between a NMDA receptor antagonist, AP-5 and an AMPA receptor antagonist, YM 872 in antinociception in the spinal cord

Background: The intrathecal N -methyl- d -aspartate (NMDA) receptor antagonist, AP-5 and the α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptor antagonist, YM 872 showed inhibition on both acute and facilitated nociception in our previous study. The present study was performed to investigate the interaction between intrathecal AP-5 and YM 872 in antinociception for acute and chronic nociception.
Methods: Sprague–Dawley rats with lumbar intrathecal catheters were tested for their thermal tail withdrawal response and for their paw flinches by formalin injection after intrathecal administration of AP-5 or YM 872. The effects of the combination were tested by an isobolographic analysis using 50% effective dose (ED50). Total fractional dose was calculated as (ED50 dose of AP-5 in combination)/(ED50 dose of AP-5 alone)+(ED50 dose of YM 872 in combination)/(ED50 dose of YM 872 alone).
Results: Intrathecally administered AP-5, YM 872, and their combination produced dose-dependent increases of the tail-flick latency and decreases in the number of flinches in both phase 1 and 2 of the formalin test. The ED50 values of the combination were significantly lower than the calculated additive values ( P <0.01). Total fractional dose value was 0.22 in the tail flick test, 0.12 in the phase 1 and 0.14 in the phase 2 of the formalin test.
Conclusion: An NMDA receptor antagonist, AP-5 and an AMPA receptor antagonist, YM 872 had synergistic antinociceptive effects on both acute thermal and inflammatory induced acute and facilitated nociception.     

NMDA受体拮抗剂LY274614对吗啡耐受性和依赖性的抑制作用

目的 观察竞争性N－甲基－D－天门冬氨酸（NMDA）受体拮抗剂LY274614对大鼠吗啡耐受性和依赖性的抑制作用。方法 对大鼠皮下注射吗啡（15mg/kg,3次／d)产生吗啡耐受性和依赖性，并在注射吗啡的同时皮下注射LY274614（2，4，6mg/kg),测定1h后大鼠热辐射甩尾反应潜伏期（TF)以评价吗啡的镇痛效果。实验的第10d，观察皮下注射纳络酮诱发的戒断症状以评价大鼠对吗啡的躯体依赖行为。结果 LY27461本身不产生镇痛作用，但与吗啡合用时可抑制吗啡耐受性产生而引起的TF下降，并可减少纳络酮诱发的戒断症状发生次数。结论 竞争性NMDA受体拮抗剂LY274614可抑制大鼠经持续皮下注射吗啡而产生的吗啡耐受性和依赖性。     

The effects of an AMPA receptor antagonist on the neurotoxicity of tetracaine intrathecally administered in rabbits

We have reported that large concentrations of intrathecal local anesthetics increase glutamate concentrations in the cerebrospinal fluid (CSF) and cause neuronal injury in rabbits. In the current study we determined whether an alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptor antagonist, YM872, administered intrathecally, reduces neuronal injury caused by tetracaine. We first examined the effects of intrathecal YM872 10, 30, 100, or 300 mug in rabbits (n = 3 in each). YM872 produced reversible motor and sensory block in a dose-dependent manner. Then, we evaluated modulatory effects of YM872 (300 mug) on tetracaine-induced glutamate release and neuronal injury. Pretreatment of YM872 did not attenuate 1% or 2% tetracaine-induced increases in cerebrospinal fluid glutamate concentrations (n = 3 in each). For evaluation of neuronal injury, rabbits were assigned to 4 groups (n = 6 in each) and intrathecally received 1% tetracaine and saline (1%T), 1% tetracaine and YM872 (1%TY), 2% tetracaine and saline (2%T), or 2% tetracaine and YM872 (2%TY). The volume of saline, YM872, and tetracaine was 0.3 mL. Saline or YM872 was administered 30 min before tetracaine administration. Neurological and histopathological assessments were performed 1 wk after the administration. Two and 1 animals respectively, showed motor and sensory dysfunction in 1%T, whereas 5 animals showed both motor and sensory dysfunction in 2%T. YM872 improved 2% tetracaine-induced motor dysfunction and neuronal damage (chromatolytic neurons, identified by round-shaped cytoplasm with loss of Nissl substance from the central part of the cell and eccentric nuclei). In 2%TY, 3 animals showed normal motor function and 3 showed mild dysfunction (ability to hop, but not normally), whereas 4 animals showed moderate dysfunction (inability to hop) in 2%T (P = 0.042). Only 2 animals showed one chromatolytic neuron in 2%TY, whereas 5 animals showed 4-16 chromatolytic neurons in 2%T (P = 0.020). These results suggest that AMPA receptor activation is involved, at least in part, in the tetracaine-induced neurotoxicity in the spinal cord.     

The effect of the selective NMDA receptor antagonist traxoprodil in the treatment of traumatic brain injury

Traumatic brain injury (TBI) remains a major public health problem, and there is a great medical need for a pharmacological treatment that could improve long-term outcome. The excitatory neurotransmitter, glutamate, has been implicated in processes leading to neurodegeneration. Traxoprodil (CP-101,606) is a novel and potent glutamate receptor antagonist that is highly selective for the NR2B subunit of the NMDA receptor; it has been shown to be neuroprotective in animal models of brain injury and ischemia. A randomized, double-blind, placebo-controlled study was therefore conducted to assess the efficacy and safety of a 72-h infusion of traxoprodil compared to placebo in subjects with computed tomography scan evidence of severe TBI (GCS 4-8). A total of 404 males and non-pregnant females, aged 16-70, were treated within 8 h of injury. At baseline, subjects were stratified by motor score severity. The results showed that a greater proportion of the traxoprodil-treated subjects had a favorable outcome on the dichotomized Glasgow Outcome Scale (dGOS) at 6 months (delta 5.5%, OR 1.3, p = 0.21, 95% CI:[0.85, 2.06]) and at last visit (delta 7.5%, OR 1.47, p = 0.07, 95% CI:[0.97, 2.25]). The mortality rate with traxoprodil treatment was 7% less than with placebo treatment (OR 1.45, p = 0.08, 95% CI:[0.96, 2.18]). Differences between treatment groups were more pronounced in the severest subset (delta 11.8% for the dGOS at last visit and delta 16.6% for mortality). Traxoprodil was well tolerated. Although these results are intriguing, no definitive claim of efficacy can be made for traxoprodil for the treatment of severe TBI.     

The effect of the preemptive use of the NMDA receptor antagonist dextromethorphan on postoperative analgesic requirements

Both central sensitization after peripheral tissue injury and the development of opiate tolerance involve activation of N-methyl-D-aspartate receptors. In this double-blinded, randomized study, we investigated the preemptive versus postincisional effects of dextromethorphan, an N-methyl-D-aspartate receptor antagonist, on postoperative pain management. Sixty ASA I and II patients undergoing elective upper abdominal surgery were randomly allocated to three equally sized groups. The Preincisional group patients received dextromethorphan (120 mg) IM 30 min before skin incision and a placebo (isotonic saline) 30 min before the end of surgery. The Postincisional group received the same dose of dextromethorphan 30 min before the end of surgery and a placebo 30 min before skin incision, and the Control group received a placebo both 30 min before skin incision and 30 min before the end of surgery. A standard general anesthetic technique including fentanyl, propofol, isoflurane, and atracurium was used. Postoperative meperidine patient-controlled analgesia (PCA) was used. There were no significant group differences in the median pain scores except in the visual analog scale at 6 h both at rest and on movement; these were significantly lower in the Preincisional group than the other two groups (P < 0.05). The mean time to initiation of PCA was significantly longer in the Preincisional than in the Postincisional and Control groups (mean [SD]: 10.7 [2.2 h], 5.4 [2.1 h], and 3.7 [1.6 h], respectively; P < 0.001]. The 24-h PCA-meperidine consumption was significantly less in the Preincisional than in the Postincisional and Control groups (mean [SD]: 140 [60 mg], 390 [80 mg], and 570 [70 mg], respectively; P < 0.001]. The incidence of postoperative hypoxemia (SpO(2) < 90%) and nausea was significantly less in the Preincisional group (P < 0.05). In conclusion, preincisional IM 120 mg dextromethorphan compared with the same postincisional dose significantly reduced postoperative meperidine consumption. IMPLICATIONS: IM administration of preincisional dextromethorphan (120 mg), allowing the use of a larger dose sufficient to block the central sensitization caused by activation of the N-methyl-D-aspartate receptors, provides preemptive analgesia and has a supportive role in postoperative pain relief, as shown by a significant decrease in 24-h meperidine consumption.     

Intrathecal co-administration of NMDA antagonist and NK-1 antagonist reduces MAC of isoflurane in rats

BACKGROUND: Intravenous administration of N-methyl-D-aspartate (NMDA) receptor antagonists and alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptor antagonists reportedly reduce the minimum alveolar anaesthetic concentration (MAC) for inhalation anaesthetics. If pain perception can be prevented by the intrathecal administration of antinociceptive receptor antagonists, these agents may reduce the requirements for inhalation anaesthetics. We studied the effect of intrathecal administration of an AMPA/kainate receptor antagonist, a metabotropic glutamate (mGlu) receptor antagonist and co-administration of NMDA and a neurokinin-1(NK-1) receptor antagonist drugs at low doses on the MAC. METHODS: After Wistar rats (n=36) were fitted with indwelling intrathecal catheters, the MAC of isoflurane was determined following intrathecal administration of a non-NMDA receptor antagonist (CNQX) at 10 microg, a mGlu receptor antagonist (AP3) at 10 microg, or a combination of NMDA receptor antagonist (APV) at 0.01 microg to 1 microg with NK-1 receptor antagonist (CP96345, CP) at 0.1 microg to 10 microg. Subsequently, a reversal dose of intrathecal NMDA with substance P (SP) was administered, and the MAC of isoflurane was redetermined. Conscious rats (n=15) were also examined for the presence of locomotor dysfunction following the intrathecal co-administration of APV and CP. RESULTS: Neither CNQX nor AP3 reduced the MAC of isoflurane. APV at 0.01 microg plus CP at 1 microg, as well as APV at 0.1 microg plus CP at 10 microg, reduced the MAC of isoflurane, with respective reductions of 7.6% and 14%; (P<0.05). Co-administration of NMDA plus SP reversed the decrease in the MAC of isoflurane. Locomotive activity was not changed. CONCLUSIONS: The NMDA receptor and the NK-1 receptor are important determinants of the MAC of isoflurane, exerting this influence by inhibition of pain transmission in the spinal cord, while mGlu and AMPA receptors have no effect on the MAC of isoflurane.     

The N-Methyl-D-aspartate (NMDA) receptor antagonist dizocilpine (MK-801) suppresses enfturane-induced opisthotonus in mice

We determined whether enflurane-induced opisthotonus in ddN mice is mediated by N-methyl-D-aspartate (NMDA) receptor using NMDA receptor antagonists dizocilpine (MK-801) and ketamine. Animals were given intraperitoneal injections of 0.2ml saline (control), 2.5 or 5.0mg·kg^–1 dizocilpine in saline, or 20 or 40mg·kg^–1 ketamine is saline 20min prior to exposure to 2.0% enflurane. Incidence of opisthotonus measured during exposure to enflurane for 20min was 49% (n = 51) in saline (control) group, 6.7 (P 0.01 vs control, n = 30) and 15.0% (P 0.01, n = 40) in 2.5 and 5.0mg·kg^–1 dizocilpine group, respectively, and 43.9 (NS, n = 41) and 40.0% (NS, n = 40) in 20 and 40mg·kg^–1 ketamine group, respectively. These results strongly suggest that enflurane-induced opisthotonus is mediated by NMDA receptor. Ketamine failed to suppress significantly due to possibly small dosages. Further, dizocilpine itself produced severe seizures during preenflurane period (30.0 and 40.0% in 2.5 and 5.0mg·kg^–1, respectively), which may be a novel finding.(Komatsu H, Nogaya J, Anabuki D, et al.: The N-methyl-D-aspartate (NMDA) receptor antagonist dizocilpine (MK-801) suppresses enflurane-induced opisthotonus in mice. J Anesth 7: 519–522, 1993)     

Spinal neurotoxicity and tolerance after repeated intrathecal administration of YM 872, an AMPA receptor antagonist,in rats

Purpose Although the -amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptor antagonist, YM 872, has been considered to be useful in analgesia for both acute and chronic pain, there are no studies of its neurotoxicity and tolerance. We examined the spinal neurotoxicity and tolerance of YM 872 analgesia by repeated intrathecal administration in rats.Methods Male Sprague-Dawley rats with lumbar intrathecal catheters received YM 872 at 1µg·10µl^–1 (eight rats; YM group) or normal saline 10µl (eight rats; C group) intrathecally once a day for 30 days. We evaluated the analgesic effects every 3 days, by tail-flick test and behavioral side effects. On the 31st day, the lumbar spinal cord was removed from four randomly selected rats in each group for histological examination.Results The YM group showed significantly longer tail-flick latency when subjected to a high-intensity light beam than the C group at each measurement time point, although no significant changes in the latency according to the time course of the study were observed for the entire study period of 30 days in either group. No rats showed any side effects. Histologically, only slight lymphocytic cell infiltration and degeneration of myelinated fibers occurred, similarly in both groups. No changes were observed in the spinal cord in either group.Conclusion Administration of YM 872 (1µg) once a day for 30 days did not induce any tolerance and caused no histological changes in the spinal cord.     

NMDA receptor antagonist felbamate reduces behavioral deficits and blood-brain barrier permeability changes after experimental subarachnoid hemorrhage in the rat

Increased levels of glutamate and aspartate have been detected after subarachnoid hemorrhage (SAH) that correlate with neurological status. The NMDA receptor antagonist felbamate (FBM; 2-phenyl-1,3-propanediol dicarbamate) is an anti-epileptic drug that elicits neuroprotective effects in different experimental models of hypoxia-ischemia. The aim of this dose-response study was to evaluate the effect of FBM after experimental SAH in rats on (1) behavioral deficits (employing a battery of assessment tasks days 1-5 post-injury) and (2) blood-brain barrier (BBB) permeability changes (quantifying microvascular alterations according to the extravasation of protein-bound Evans Blue by a spectrophotofluorimetric technique 2 days post-injury). Animals were injected with 400 muL of autologous blood into the cisterna magna. Within 5 min, rats received daily oral administration of FBM (15, 30, or 45 mg/kg) for 2 or 5 days. Results were compared with sham-injured controls treated with oral saline or FBM (15, 30, or 45 mg/kg). FBM administration significantly ameliorated SAH-related changes in Beam Balance scores on days 1 and 2 and Beam Balance time on days 1-3, Beam Walking performance on days 1 and 2, and Body Weight on days 3-5. FBM also decreased BBB permeability changes in frontal, temporal, parietal, occipital, and cerebellar cortices; subcortical and cerebellar gray matter; and brainstem. This study demonstrates that, in terms of behavioral and microvascular effects, FBM is beneficial in a dose-dependent manner after experimental SAH in rats. These results reinforce the concept that NMDA excitotoxicity is involved in the cerebral dysfunction that follows SAH.     

The NMDA NR2B subunit-selective receptor antagonist, CP-101,606, enhances the functional recovery the NMDA NR2B subunit-selective receptor and reduces brain damage after cortical compression-induced brain ischemia

Using a novel in vivo model for cerebral ischemia produced by short-lasting compression of a well-defined brain area of sensorimotor cortex we studied neuroprotective effects of the NMDA NR2B subunit selective antagonist, CP-101,606, in Sprague-Dawley rats. Cortical compression for 30 min produced a consistent and highly reproducible functional impairment, that is paresis of contralateral hind and fore limbs. The neurological deficit was accompanied by marked brain damage in cerebral cortex, hippocampus and thalamus as identified by Fluoro-Jade, a marker of general neuronal cell death. Using a daily performed beam walking test it was shown that untreated animals recovered from their functional impairment within 5-7 days following surgery. Intravenous administration of increasing doses (1, 5, 10, 20 mg/kg) of the NMDA NR2B subunit receptor specific antagonist, CP-101,606, dose-dependently improved the rate of functional recovery and protected against the ischemic brain damage in cerebral cortex, hippocampus, and thalamus as identified 2 days after the ischemic insult. Based upon these results, we conclude that NMDA NR2B receptor subunits represent potential targets to reduce not only the functional deficits, but also neuronal death in cortex and several midbrain regions produced by moderate, transient, cerebral ischemia.     

Anti-cholinesterase activity of dopamine D2 receptor antagonist: its clinical significance

Anti-cholinesterase activity of dopamine D2 receptor antagonist, domperidone was studied by means of chronically implanted force transducers in the gastrointestinal (GI) tract in five conscious dogs. Cisapride was used as a drug to stimulate endogenous release of acetylcholine. In the digestive state, cisapride (0.25 mg/kg) stimulated 18.6 +/- 5.6% increase in the motor index of the gastric antrum alone, however, combined administration with domperidone (1.0 mg/kg-hr) significantly enhanced the motor index in the gastric antrum and duodenum. In the gastric antrum, the increase was 68.1 +/- 7.2%. During the interdigestive state, cisapride did not always induce the interdigestive migrating contractions (IMC)-like contractions in the GI tract, but the background infusion of domperidone significantly increased the incidence of the occurrence of IMC-like contractions by cisapride. In in vitro study, weak but significant anti-cholinesterase activity was found in domperidone, the activity being about 1/1,000 of that of neostigmine. In dog experiment, similar enhancement of motor stimulating activity of cisapride was observed when neostigmine was given at 1.0 micrograms/kg-hr. In conclusion, domperidone has anti-cholinesterase activity and acts to enhance motor stimulating activity of cisapride through inhibition of cholinesterase activity in the upper digestive tract.     

异氟醚和NMDA拮抗剂AP5与脑皮层谷氨酸含量变化的研究

目的研究单纯异氟醚及相同MAC下异氟醚复合NMDA拮抗剂AP5麻醉时,脑皮层谷氨酸(G)含量的变化.方法6只成年雄性大鼠持续吸入异氟醚,脑皮层内插入0.05mm内径的微透析探针,用人工脑脊液灌注,测定基础MAC后,分为四个浓度组,Ⅰ组异氟醚吸入浓度1.0%,Ⅱ组浓度1MAC,Ⅲ组浓度2.096,每一浓度平衡20min后,收集透析液,并与定量脑电图(EEG)和体感诱发电位(SEP)监测同步;上述步骤进行完后,经尾静脉缓慢注射AP5120mg/kg+持续泵注100mg@kg-1@h-1,给药后30min再测MAC,测得的MAC记为AP5-MAC,即Ⅳ组,余处理同前.用高压液相仪测定微透析液样品中G的水平.结果异氟醚吸入浓度与G水平呈明显负相关(r=-0.839,P＜0.01).Ⅱ组与Ⅳ组比较,达1MAC时Ⅳ组所需吸入浓度降低,G含量下降,EEG97%谱边界频率(97%SEF)显著下降,SEP潜伏期延长,振幅无统计学差异.结论异氟醚可使鼠脑皮层兴奋性递质谷氨酸含量呈剂量依赖性下降,静注NMDA拮抗剂AP5可减少异氟醚的MAC及皮层谷氨酸含量,增强异氟醚的麻醉效能,说明异氟醚可能通过突触前机制,产生兴奋性神经传递作用的抑制.     

Agonist and antagonist NMDA receptor effect on cell fate during germ cell differentiation and regulate apoptotic process in 3D organ culture

In this work, agonist and antagonist N-methyl-D-aspartate (NMDA) receptor activation effect on cell fate during germ cell differentiation and regulate apoptotic process in 3D organ culture were studied. Afterwards, the effect of D-serine, retinoic acid (RA) and MK801 on spermatogenesis development was investigated. The animals were injected a single dose (40 mg/kg, intraperitoneal) of busulfan. After confirming the model, ten 5-day-old NMRI mice were used as spermatogonial stem cells (SSCs) transplantation donors. The SSCs were confirmed by detecting the promyelocytic leukaemia zinc finger (PLZF) protein. Then, tissue culture of the azoospermia model which had received SSCs was performed in various conditions (seven groups). The apoptosis markers levels of cells were significantly decreased in differentiation media containing RA and serine. In contrast, the expression of apoptotic markers including caspase 3, caspase 9 and Bax was increased in the presence of MK801. In conclusion, a new in vitro system capable of producing mature spermatozoa was developed that would be useful for investigating the medicinal effects of agents on the male reproductive system. Also, a comparison of spermatogenesis development in different media revealed that the presence of D-serine and RA (retinoic acid) in the culture medium has a positive effect on spermatogenesis.     

NPS 1506: a novel NMDA receptor antagonist: neuroprotective effects in a model of closed head trauma in rats

We examined whether NPS 1506, a novel uncompetitive N-methyl-D-aspartate receptor antagonist, influences neurological outcome following closed head trauma (CHT) in rats. One hundred ten rats were divided into 11 groups: CHT (yes/no), treatment with NPS 1506 (yes/no), and time of euthanization (24 h/48 h). The dose of NPS 1506 was 1 mg/kg IV at 1 and 4 hours following CHT or sham operation. Closed head trauma induced the following changes in the injured hemisphere: Decreased specific gravity (sg) (1.036 +/- 0.006) and magnesium (Mg) (0.042 +/- 0.005 microg/mg) at 24 hours, and potassium (K) at 24 (1.145 +/- 0.376 microg/mg) and 48 hours, and increased water content (W) (84.9 +/- 2.5%) and sodium (Na) (2.135 +/- 0.699 microg/mg) at 24 hours, and calcium (Ca) at 24 (0.543 +/- 0.157 microg/mg) and 48 hours. These were reversed by NPS 1506; sg of 1.043 +/- 0.004, Mg of 0.077 +/- 0.009 microg/mg, K of 1.930 +/- 0.238 microg/mg, W of 81.5 +/- 1.9%, Ca of 0.043 +/- 0.023 microg/mg, and Na of 0.688 +/- 0.110 microg/mg. In groups not given NPS 1506, a nonsignificant decrease in neurological severity score (NSS) occurred at 24 and 48 hours as compared to NSS at 1 hour after CHT. In groups given NPS 1506, NSS at 24 and 48 hours decreased significantly (improved) compared to NSS at 1 hour, but not compared to NSS at 24 and 48 hours in groups not given NPS 1506. NPS 1506 caused no significant change in ischemic tissue volume or hemorrhagic necrosis volume in the injured hemisphere at 24 hours or 48 hours. These findings indicate that NPS 1506 improved measures of brain tissue edema (at 24 hours but not at 48 hours) and ion homeostasis, and this improvement was not related to other measures of outcome.     

CSF and plasma pharmacokinetics of the NMDA receptor antagonist CPP after intrathecal, extradural and i.v. administration in anaesthetized pigs

The N-methyl-D-aspartate (NMDA) receptor complex plays a centraldegrees in the modulation of neuronal information in the centralnervous system. This study was designed to examine the pharmacokineticsof the NMDA antagonist 3-(2-carboxypiperazin-4-yl)propyl-1-phosphonicacid (CPP) in plasma and cerebrospinal fluid (CSF) and rostralspread in the CSF after lumbar intrathecal, extradural and i.v.administration. Anaesthetized pigs were given a lumbar intrathecal,lumbar extradural or an i.v. injection of a mixture of [³H]labelledand unlabelled CPP. CSF was sampled over 10 h through intrathecalcatheters positioned at the L1, T5 and C1 vertebral levels.Blood samples were obtained over the same period. Haemodynamicand arterial blood-gas variables and acid-base balance weremonitored during the study. The area under the radioactivityconcentration-time curves showed a gradient between cervicaland lumbar CSF radioactivity of about 1:2500 after intrathecaladministration and about 1:140 after extradural administration,indicating that only small fractions of lumbar administeredCPP spread rostrally. About 2% of an extradurally administereddose was found in the CSF. After i.v. administration of [³H]CPP,clearance was mean 122 (SEM 16) ml min^–1 and the CSF:serumradioactivity gradient was approximately 1:4. The half-lifeof [³H]CPP varied little (mean range 94–191 min) irrespectiveof the route of administration or the level of sampling. Cervicalradioactivity after lumbar intrathecal administration probablyresulted from rostral transport via CSF bulk flow, whereas afterextradural administration, systemic absorption and redistributionvia the blood-brain barrier probably contributed. Renal excretionwas the main route of systemic elimination. No effects on haemodynamics,arterial blood-gas tensions or acid-base balance could be correlatedwith intrathecal or extradural administration of CPP. The steepgradient between cervical and lumbar concentrations of [³H]CPPsuggests that it may be possible to administer CPP spinallyat the lumbar level in pharmacologically active doses with littledistribution to the supraspinal level.