Theoretical Additive Values Research Articles

Combination of a δ-opioid Receptor Agonist and Loperamide Produces Peripherally-mediated Analgesic Synergy in Mice.

The long-term use of opioids for analgesia carries significant risk for tolerance, addiction, and diversion. These adverse effects are largely mediated by μ-opioid receptors in the central nervous system. Based on the authors' previous observation that morphine and δ-opioid receptor agonists synergize in spinal cord in a protein kinase Cε-dependent manner, they predicted that this μ-opioid receptor-δ-opioid receptor synergy would take place in the central terminals of nociceptive afferent fibers and generalize to their peripheral terminals. Therefore, the authors hypothesized that loperamide, a highly efficacious μ-opioid receptor agonist that is excluded from the central nervous system, and oxymorphindole, a δ-opioid receptor agonist that was shown to synergize with morphine spinally, would synergistically reverse complete Freund's adjuvant-induced hyperalgesia. Using the Hargreaves assay for thermal nociception, the von Frey assay for mechanical nociception and the complete Freund's adjuvant-induced model of inflammatory pain, we tested the antinociceptive and antihyperalgesic effect of loperamide, oxymorphindole, or the loperamide-oxymorphindole combination. Animals (Institute for Cancer Research [ICR] CD1 strain mice; n = 511) received drug by systemic injection, intraplantar injection to the injured paw, or a transdermal solution on the injured paw. Dose-response curves for each route of administration and each nociceptive test were generated, and analgesic synergy was assessed by isobolographic analysis. In naïve animals, the loperamide-oxymorphindole combination ED50 value was 10 times lower than the theoretical additive ED50 value whether given systemically or locally. In inflamed animals, the combination was 150 times more potent systemically, and 84 times more potent locally. All combinations showed statistically significant synergy when compared to the theoretical additive values, as verified by isobolographic analysis. The antihyperalgesia was ablated by a peripherally-restricted opioid antagonist. From these data we conclude that the loperamide-oxymorphindole combination synergistically reverses complete Freund's adjuvant-induced inflammatory hyperalgesia. The authors also conclude that this interaction is mediated by opioid receptors located in the peripheral nervous system.

Synergistic antinociceptive interactions between fospropofol and alfentanil in mice

BackgroundCombination therapy, which provides the opportunity to achieve optimal analgesia with reduced side effects at lower drug doses, is a valid approach for the treatment of pain. The analgesic interaction between fospropofol and alfentanil has not been investigated till date. We sought to determine the nature of the interaction between fospropofol and alfentanil in mice models of the formalin test, hot-plate test and the tail-flick test. MethodsThe effects of fospropofol, alfentanil and their combinations were examined in the formalin-induced paw inflammatory hyperalgesia, the hot-plate test and the tail-flick test in mice. In the three models, dose–response curves were established and their respective ED50 (50% effective dose) values were determined separately for each agent. Fixed-ratio combinations of fospropofol and alfentanil were tested for their combined antinociceptive effects, and the type of interaction was determined by the isobolographic analysis. ResultsFospropofol, alfentanil and their combination produced a dose-dependent decrease in the number of flinches during phase 1 of the formalin test. In the hot-plate test and in the tail-flick test, fospropofol, alfentanil and their combination significantly and dose dependently prolonged the latency of withdrawal. In the three models, isobolographic analysis revealed a significant synergistic interaction between fospropofol and alfentanil. The ED50 value for the drug combination was significantly lower than the theoretical additive value (p<0.05). ConclusionsThe results demonstrate that fospropofol and alfentanil provide synergistic antinociceptive interactions in the formalin, hot-plate, and tail-flick tests. The observed synergistic interaction between fospropofol and alfentanil are indicative of the effectiveness of the combination treatment in pain management and should be explored further in patients undergoing minor surgical procedures.

Antinociceptive drug interaction between intrathecal vitamin E and gabapentin in the rat formalin test

BackgroundGabapentin is thought to exert an effect through the voltage-dependent calcium channel. Vitamin E is a widely known antioxidant which neutralizes the harmful effect of ROS which is considered to play a prominent role in various painful conditions. This study was therefore conducted to assess the antinociceptive effects of gabapentin and vitamin E and the interaction of these drugs in the modulation of pain in rats subjected to a formalin test.MethodsSprague-Dawley rats with a lumbar intrathecal catheter were tested for their paw flinches by 5% formalin injection after intrathecal injection of gabapentin or vitamin E. After obtaining dose-response curves for each drug, the effect of the combination was tested by the total dose fraction value and isobolographic analysis.ResultsWhen a single drug was injected intrathecally, significant dose-dependent decreases in flinches were shown only in the late phase. ED50 values of intrathecal gabapentin and vitamin E in the late phase were 75.3 ± 9.58 µg, and 17.56 ± 1.65 mg/kg respectively. The combination of gabapentin and vitamin E produced dose-dependent decreases in the number of flinches in both phases induced by the formalin test. The ED50 value of the combination was lower than the theoretical additive values in the late phase, but did not show a significant difference with the theoretical additive value.ConclusionsGabapentin and vitamin E (by itself) have no antinociceptive effect in the early phase; however their combination has shown an antinociceptive effect. In addition, they show additive effects in the late phase of the formalin test.

S51 Thioredoxin modifies MIF release from human monocytes following stimulation with LTA and LPS

RationaleThioredoxin (Trx) is a 12-kDa ubiquitous redox-active thiol (-SH) protein. Plasma levels of Trx are raised in numerous medical and surgical conditions associated with oxidative stress and inflammation such as...

Interaction between midazolam and epibatidine in spinally mediated antinociception in rats

Both gamma-aminobutyric acid (GABA)(A) receptors and nicotinic cholinergic receptors have important roles in antinociception in the spinal cord. The antinociceptive effects of midazolam (a GABA(A) agonist) and epibatidine (a nicotinic cholinergic agonist) in the spinal cord have been reported. The present study was performed to investigate the interaction between intrathecal midazolam and epibatidine. Sprague-Dawley rats with lumbar intrathecal catheters were tested for their tail withdrawal response to thermal stimulation (tail-flick test) or for their paw flinches in response to formalin injection (formalin test) after the intrathecal administration of epibatidine or a combination of midazolam and epibatidine. The combination doses were 1/16, 1/8, 1/4, 1/2, 1, 2, and 4 times the 50% effective dose (ED50) of each agent in each test. The interaction of midazolam and epibatidine was investigated by isobolographic analysis. Behavioral side effects were also investigated. In the tail-flick test, the ED50 values of the combination were significantly higher than the theoretical additive values. In the formalin test, the ED50 values of the combination were significantly lower than the theoretical additive values in phase 1, but were not different from the theoretical additive values in phase 2. The intrathecal combination of midazolam and epibatidine had antagonistic effects on thermal acute nociception, while the combination had synergistic effects on acute inflammatory nociception, with only additive effects on inflammatory-facilitated nociceptive responses.

Synergy between a NR2B receptor antagonist DHQ and 3-methyl-gabapentin in mice with neuropathic pain

The synergistic effect of a selective NR2B NMDA receptor antagonist, (−)-( R)-6-{2-[4-(3-Fluorophenyl)-4-hydroxy-1-piperidinyl]-1-hydroxyethyl-3,4-dihydro-2(1 H)-quinolinone (DHQ), and a α 2δ ligand, 3-methyl-gabapentin (3M-GBP), was investigated in the mouse partial sciatic nerve model. The interaction was observed after administration of DHQ and 3M-GBP combination at fixed dose ratios of 1:10 and 1:30 and the dose–response curves shifted approximately 13- and 17-fold leftward, respectively, from the theoretical additive values. However, a fixed dose ratio of 1:50 resulted only in an additive effect. These results indicate the synergistic interaction between DHQ and 3M-GBP in this animal model of neuropathic pain.

Mixtures of pregelatinised maize starch and κ-carrageenan: Compatibility, rheology and gelation

Compatibility, flow and visco-elastic properties of a pregelatinised maize starch mixed with κ-carrageenan were investigated. After cooking of the pregelatinised starch, some undissolved granules remained in solution. Aqueous mixtures of κ-carrageenan and starch were studied at 60 °C and 20 °C by combining rheological measurements and microscopic observations under conditions allowing gelation of carrageenan and non-gelation of starch. The viscometric study of mixed dilute solutions of amylose from pregelatinised starch and carrageenan showed that the components are slightly incompatible. Mixture viscosity and elastic modulus were studied at 60 °C in details as a function of mixture composition for a total polymer concentration of 3%; both were found to be significantly higher than the corresponding theoretical additive values. This finding was interpreted by starch granules excluded volume effect. At 20 °C, no noticeable increase of mixture elastic modulus was found as compared with the additive value. The absence of the synergistic effect is supposed to be due to the formation of highly inhomogeneous gels with agglomerates of undissolved granules.

Antinociceptive Synergy Between the Cannabinoid Receptor Agonist WIN 55,212-2 and Bupivacaine in the Rat Formalin Test

The analgesic interaction between cannabinoids and local anesthetics has not been investigated. We sought to determine the nature of the interaction between the intrathecal cannabinoid receptor agonist (WIN 55,212-2) and bupivacaine using the formalin test. Lumbar intrathecal catheters were implanted in male Sprague-Dawley rats. After intrathecal administration of WIN 55,212-2, bupivacaine, or their combination, 50 microL of 5% formalin was injected subcutaneously into the hindpaw. Dose-response curves were established and the respective ED50 (50% effective dose) values were determined for each agent alone. Fixed-ratio combinations of WIN 55,212-2 and bupivacaine were tested for combined antinociceptive effects in the formalin test and an isobolographic analysis was performed to characterize the pharmacologic interaction of both drugs. Intrathecally administered WIN 55,212-2, bupivacaine, or their combination produced a dose-dependent decrease in the number of flinches during Phase 1 and 2 of the formalin test. Isobolographic analysis revealed a synergistic interaction between intrathecal WIN 55,212-2 and bupivacaine in both phases of the formalin test. In combination, the ED50 value was significantly smaller than the theoretical additive value (P < 0.05). These results demonstrate that intrathecally coadministered WIN 55,212-2 and bupivacaine provide synergistic antinociceptive interaction in both phases of the formalin test.

The Interaction between Intrathecal NMDA Receptor Antagonist and 5-HT3Receptor Agonist in the Rat Formalin Test

Background: A single agent administered is not effective enough to block nociception. Combination of agents acting through different mechanisms may be one of the best ways for better analgesic methods. The purpose of this study was to investigate the spinally mediated analgesic interaction between 5-HT3 receptor agonist (m-CPBG) and NMDA receptor antagonist (MK801) in the rat formalin test. Methods: A polyethylene catheter was inserted in the atlanto-occipital membrane to the thoracolumbar level of the spinal cord. On postoperative 7 day, the normal saline (n = 6), m-CPBG (10, 30, 100μg: n = 18) and MK801 (0.1, 1, 10μg: n = 18) were injected intrathecally before subcutaneous injection of 5% formalin. The combinations of each 1/2ED50 + 1/2ED50, 1/4ED50 + 1/4ED50, 1/8ED50 + 1/8ED50 were tested and the ED50 of the combination was determined. The experimental ED50 values were compared with the theoretical additive values. Results: m-CPBG dose-dependently decreased the number of flinches in both phase 1 and 2. MK801 dose-dependently reduced the number of flinches in phase 2, but not in phase 1. The combination of m-CPBG and MK801 produced dose-dependent decreases of flinches on both phase 1 and 2. ED50 values (2.75μg for m-CPBG and 0.076μg for MK801) of phase 2 in the combination were significantly less than the calculated additive values (20.0μg for m-CPBG and 0.55μg for MK801)(P ＜ 0.01). Conclusions: Intrathecally administered m-CPBG and MK801 had synergistic analgesic effects on formalin-induced hyperalgesia as well as antinociceptive effects in phase 1 on the rat formalin test.

Antihyperalgesic Effects of Intrathecal Gabapentin and CNQX, Non-NMDA Receptor Antagonist, in a Rat Model for Postoperative Pain

Background: Although the descriptions of the pharmacological action mechanisms of gabapentin remain incomplete, gabapentin has been proven to provide analgesic effects in inflammatory, bum injury and neuropathic pain models. However, the analgesic effect of gabapentin in postoperative pain has not been demonstrated. Of the antagonists of the excitatory neurotransmitter receptor in the spinal dorsal horn, only the antagonists of the non-NMDA receptor have proven effective in a postoperative acute pain model. In the current study, the antihyperalgesic effects of intrathecal gabapentin and CNQX (6-cyano-7-nitroquinoxaline-2, 3-dione) were determined in a rat model of postoperative pain. Furthermore, the drug interaction when both drugs were co-administrated was also tested. Methods: A lumbar intrathecal catheter was implanted into Sprague-Dawley rats (weight 300-350 g) and the rats then assigned to receive one of several doses of gabapentin (10, 30, 100, 300 ), CNQX (1, 5, 10, 25 ) and gabapentin + CNQX (1/2 for each drug, 1/4 , 1/8 ), and the level of withdrawal threshold to the von Frey Filament near the incision site determined. The dose response curves were obtained, and the ED50 of each drug calculated. Isobolographic and fractional analyses were performed to study the drug interaction. Results: Both gabapentin and CNQX displayed dose dependent antihyperalgesic effects. The for gabapentin and CNQX were 56.2 (95% confidence interval [CI], 45.3 -69.7 ) and 4.6 (CI, 3.2 -6.5 ), respectively. When both drugs were co-administrated, the for gabapentin and CNQX were 23.4 (CI, 20.2 - 27.1 ) and 1.6 (CI, 1.3 -1.8 ), respectively. In the fractional analysis, the fraction obtained using this numeric value was 0.77. In the isobologram analysis, the obtained value of the gabapentin-CNQX mixture was below the theoretical additive value on the diagonal line, but this was not statistically significant. Conclusions: The intrathecal co-administration of gabapentin and CNQX showed an additive antihyperalgesic effect in our rat model of postoperative pain.

Gabapentin and the neurokinin(1) receptor antagonist CI-1021 act synergistically in two rat models of neuropathic pain.

The present study examines the effect of combinations of gabapentin (Neurontin) and a selective neurokinin (NK)(1) receptor antagonist, 1-(1H-indol-3-ylmethyl)-1-methyl-2-oxo-2-[(1-phenylethyl)amino]ethyl]-2-benzofuranylmethyl ester (CI-1021), in two models of neuropathic pain. Dose responses to both gabapentin and CI-1021 were performed against static allodynia induced in the streptozocin and chronic constriction injury (CCI) models. Theoretical additive lines were calculated from these data. Dose responses to various fixed dose ratios of a gabapentin/CI-1021 combination were then examined in both models. In the streptozocin model, administration of gabapentin/CI-1021 combinations at fixed dose ratios of 1:1 and 60:1 resulted in an additive effect with dose response similar to the theoretical additive line. However, a synergistic interaction was seen after fixed dose ratios of 10:1, 20:1, and 40:1 with static allodynia completely blocked and the dose responses shifted approximately 8-, 30-, and 10-fold leftward, respectively, from the theoretical additive values. In the CCI model, after fixed dose ratios of 5:1 and 20:1, combinations of gabapentin and CI-1021 produced an additive response. At the fixed dose ratio of 10:1 static allodynia was completely blocked with an approximate 10-fold leftward shift of the dose response from the theoretical additive value, indicating synergy. The combination of gabapentin with a structurally unrelated NK(1) receptor antagonist, (2S,3S)-3-(2-methoxybenzylamino)-2-phenylpiperidine (CP-99,994), also produced synergy, at a fixed dose ratio of 20:1. This ratio completely blocked streptozocin-induced static allodynia and was approximately shifted leftward 5-fold from the theoretical additive value. These data suggest a synergistic interaction between gabapentin and NK(1) receptor antagonists in animal models of neuropathic pain.

DNA-damaging effects of genotoxins in mixture: nonadditive effects of aflatoxin B1 and N-acetylaminofluorene on their mutagenicity in Salmonella typhimurium.

Most animal genotoxicity studies have used exposures to single chemicals; humans, however, are potentially exposed to mixtures of genotoxins. Cancer and developmental toxicity risks associated with genotoxins in mixture are generally estimated by assuming additivity of the components. Two or more genotoxins acting sequentially or simultaneously may present a greater or lesser hazard than that predicted by simple addition of their potencies. Previously, we studied the effect of one genotoxin on the binding of a second genotoxin to DNA in an in vitro system and demonstrated that consecutive binding of the two toxins was not additive. In the present study, the effect of one genotoxin on the mutagenicity of another was evaluated for two well-known genotoxins using the Salmonella assay. Pretreatment of frameshift strains TA98 and TA1538 with AFB1-8,9-epoxide (17.3 ng/plate) enhanced the mutagenicity induced by subsequent exposure to N-acetoxy-acetylaminofluorene (N-AcO-AAF) approximately 2-3 times above theoretical values for additivity. Pretreatment of base-substitution strain TA100 with N-AcO-AAF (0.1 microg/plate) inhibited the mutagenicity following subsequent exposure to AFB1-8,9-epoxide by 3 times below the theoretical additive value. Concentration-response relationships for these enhancing or inhibitory effects were demonstrated using increasing concentrations of the first genotoxin during pretreatment. These results demonstrate effects, other than additive, of sequential exposures to two genotoxins on the induction of mutations in a bacterial system.