Analgesic effect in rodents of native and recombinant Ph alpha 1beta toxin, a high-voltage-activated calcium channel blocker isolated from armed spider venom.
Calcium influx through neuronal voltage-sensitive calcium channels (VSCC S) mediates nociceptive information in the spinal dorsal horn. In fact, spinally administered VSCC S blockers, such as omega-conotoxin MVIIA, have analgesic effect apart of their low therapeutic index and many side effects. Here we study the analgesic potential of Ph alpha 1beta, a calcium channel blocker, in rodent models of acute and persistent pain. Spinally administered Ph alpha 1beta showed higher efficacy and long-lasting analgesia in a thermal model of pain, when compared with omega-conotoxin MVIIA. Moreover, Ph alpha 1beta was more effective and potent than omega-conotoxin MVIIA not only to prevent, but especially to reverse, previously installed persistent chemical and neuropathic pain. Furthermore, the analgesic action of both toxins are related with the inhibition of Ca2+-evoked release of pro-nociceptive neurotransmitter, glutamate, from rat spinal cord synaptosomes and decrease of glutamate overflow in cerebrospinal fluid. When side effects were assessed, we found that Ph alpha 1beta had a therapeutic index wider than omega- conotoxin MVIIA. Finally, recombinant Ph alpha 1beta expressed in Escherichia coli showed marked analgesic activity similar to the native toxin. Taken together, the present study demonstrates that native and recombinant Ph alpha 1beta have analgesic effects in rodent models of pain, suggesting that this toxin may have potential to be used as a drug in the control of persistent pathological pain.
Souza
Alessandra H
AH
Departamento de Farmacologia, ICB, UFMG, 31270-901 Belo Horizonte, MG, Brazil Departamento de Quimica UFSM, 97105-900 Santa Maria, RS, Brazil Fundação Ezequiel Dias, Belo Horizonte, MG, Brazil Departamento de Bioquimica e Imunologia, ICB, UFMG, 31270-901 Belo Horizonte, MG, Brazil Nucleo Pós Graduação, Santa Casa de Belo Horizonte, Brazil Robarts Research Institute, University of Western Ontario, London, Ontario, Canada Laboratório de Neurociência, Faculdade de Medicina, UFMG, 30130-100, Brazil.
Ferreira
Juliano
J
Cordeiro
Marta do Nascimento
MDN
Vieira
Luciene Bruno
LB
De Castro
Celio J
CJ
Trevisan
Gabriela
G
Reis
Helton
H
Souza
Ivana Assis
IA
Richardson
Michael
M
Prado
Marco A M
MAM
Prado
Vânia F
VF
Gomez
Marcus Vinicius
MV
D000700:Analgesics / Q000008:administration & dosage; D000818:Animals; D002121:Calcium Channel Blockers / Q000008:administration & dosage* / Q000302:isolation & purification*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D006930:Hyperalgesia / Q000175:diagnosis / Q000503:physiopathology* / Q000517:prevention & control*; D008297:Male; D051379:Mice; D010147:Pain Measurement / Q000187:drug effects*; D011994:Recombinant Proteins / Q000378:metabolism; D013111:Spider Venoms / Q000737:chemistry* / Q000235:genetics
D000700:Analgesics; D002121:Calcium Channel Blockers; D011994:Recombinant Proteins; D013111:Spider Venoms
Altier C, Dale CS, Kisilevsky AE, Chapman K, Castiglioni AJ, Matthews EA, et al. Differential role of N-type calcium channel splice isoforms in pain. J Neurosci. 2007;27:6363-6373.
Altier C, Zamponi GW. Targeting Ca2+ channels to treat pain: T-type versus N-type. Trends Pharmacol Sci. 2004;25:465-470.
Bowersox SS, Gadbois T, Singh T, Pettus M, Wang YX, Luther RR. Selective N-type neuronal voltage-sensitive calcium channel blocker, SNX-111, produces spinal antinociception in rat models of acute, persistent and neuropathic pain. J Pharmacol Exp Ther. 1996;279:1243-1249.
Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72:248-254.
Chaplan SR, Bach FW, Pogrel JW, Chung JM, Yaksh TL. Quantitative assessment of tactile allodynia in the rat paw. J Neurosci Methods. 1994;53:55-63.
Cordeiro Mdo N, de sueiredo SG, Valentim Ado C, Diniz CR, von Eickstedt VR, Gilroy J, et al. Purification and amino acid sequences of six Tx3 type neurotoxins from the venom of the Brazilian ‘armed’ spider Phoneutria nigriventer (Keys). Toxicon. 1993;31:35-42.
Dixon WJ. Efficient analysis of experimental observations. Annu Rev Pharmacol Toxicol. 1980;20:441-462.
Dubuisson D, Dennis SG. The formalin test: a quantitative study of the analgesic effects of morphine, meperidine, and brain stem stimulation in rats and cats. Pain. 1977;4:161-174.
Gomez MV, Kalapothakis E, Guatimosim C, Prado MA. Phoneutria nigriventer venom: a cocktail of toxins that affect ion channels. Cell Mol Neurobiol. 2002;22:579-588.
Gruner W, Silva LR. Omega-conotoxin sensitivity and presynaptic inhibition of glutamatergic sensory neurotransmission in vitro. J Neurosci. 1994;14:2800-2808.
Grynkiewicz G, Poenie M, Tsien RY. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985;260:3440-3450.
Guatimosim C, Romano-Silva MA, Cruz JS, Beirao PS, Kalapothakis E, Moraes-Santos T, et al. A toxin from the spider Phoneutria nigriventer that blocks calcium channels coupled to exocytosis. Br J Pharmacol. 1997;122:591-597.
Hylden JL, Wilcox GL. Intrathecal morphine in mice: a new technique. Eur J Pharmacol. 1980;67:313-316.
Ji RR, Woolf CJ. Neuronal plasticity and signal transduction in nociceptive neurons: implications for the initiation and maintenance of pathological pain. Neurobiol Dis. 2001;8:1-10.
Kristipati R, Nadasdi L, Tarczy-Hornoch K, Lau K, Miljanich GP, Ramachandran J, et al. Characterization of the binding of omega-conopeptides to different classes of non-L-type neuronal calcium channels. Mol Cell Neurosci. 1994;5:219-228.
Lewis RJ, Nielsen KJ, Craik DJ, Loughnan ML, Adams DA, Sharpe IA, et al. Novel omega-conotoxins from Conus catus discriminate among neuronal calcium channel subtypes. J Biol Chem. 2000;275:35335-35344.
Li X, Eisenach JC. Alpha2A-adrenoceptor stimulation reduces capsaicin-induced glutamate release from spinal cord synaptosomes. J Pharmacol Exp Ther. 2001;299:939-944.
Malmberg AB, Basbaum AI. Partial sciatic nerve injury in the mouse as a model of neuropathic pain: behavioral and neuroanatomical correlates. Pain. 1998;76:215-222.
Malmberg AB, Yaksh TL. Voltage-sensitive calcium channels in spinal nociceptive processing: blockade of N- and P-type channels inhibits formalin-induced nociception. J Neurosci. 1994;14:4882-4890.
Malmberg AB, Yaksh TL. Cyclooxygenase inhibition and the spinal release of prostaglandin E2 and amino acids evoked by paw formalin injection: a microdialysis study in unanesthetized rats. J Neurosci. 1995;15:2768-2776.
Mathur SV. Ziconotide: a new pharmacological class of drug for the management of pain. Semin Anesth. 2000;19:67-75.
Mestre C, Pelissier T, Fialip J, Wilcox G, Eschalier A. A method to perform direct transcutaneous intrathecal injection in rats. J Pharmacol Toxicol Methods. 1994;32:197-200.
Miljanich GP, Ramachandran J. Antagonists of neuronal calcium channels: structure, function, and therapeutic implications. Annu Rev Pharmacol Toxicol. 1995;35:707-734.
Nicholls DG, Sihra TS, Sanchez-Prieto J. Calcium-dependent and -independent release of glutamate from synaptosomes monitored by continuous fluorometry. J Neurochem. 1987;49:50-57.
Penn RD, Paice JA. Adverse effects associated with the intrathecal administration of ziconotide. Pain. 2000;85:291-296.
Prado MA, Guatimosim C, Gomez MV, Diniz CR, Cordeiro MN, Romano-Silva MA. A novel tool for the investigation of glutamate release from rat cerebrocortical synaptosomes: the toxin Tx3-3 from the venom of the spider Phoneutria nigriventer. Biochem J. 1996;314:145-150.
Reis HJ, Prado MA, Kalapothakis E, Cordeiro MN, Diniz CR, De Marco LA, et al. Inhibition of glutamate uptake by a polypeptide toxin (phoneutriatoxin 3–4) from the spider Phoneutria nigriventer. Biochem J. 1999;343:413-418.
Romano-Silva MA, Ribeiro-Santos R, Ribeiro AM, Gomez MV, Diniz CR, Cordeiro MN, et al. Rat cortical synaptosomes have more than one mechanism for Ca2+ entry linked to rapid glutamate release: studies using the Phoneutria nigriventer toxin PhTX2 and potassium depolarization. Biochem J. 1993;296:313-319.
Saegusa H, Kurihara T, Zong S, Kazuno A, Matsuda Y, Nonaka T, et al. Suppression of inflammatory and neuropathic pain symptoms in mice lacking the N-type Ca2+ channel. EMBO J. 2001;20:2349-2356.
Smith MT, Cabot PJ, Ross FB, Robertson AD, Lewis RJ. The novel N-type calcium channel blocker, AM336, produces potent dose-dependent antinociception after intrathecal dosing in rats and inhibits substance P release in rat spinal cord slices. Pain. 2002;96:119-127.
Snutch TP. Targeting chronic and neuropathic pain: the N-type calcium channel comes of age. NeuroRx. 2005;2:662-670.
Staats PS, Hekmat H, Staats AW. The psychological behaviorism theory of pain and the placebo: its principles and results of research application. Adv Psychosom Med. 2004;25:28-40.
Takahashi T, Momiyama A. Different types of calcium channels mediate central synaptic transmission. Nature. 1993;366:156-158.
Vieira LB, Kushmerick C, Hildebrand ME, Garcia E, Stea A, Cordeiro MN, et al. Inhibition of high voltage-activated calcium channels by spider toxin PnTx3-6. J Pharmacol Exp Ther. 2005;314:1370-1377.
Wang YX, Bezprozvannaya S, Bowersox SS, Nadasdi L, Miljanich G, Mezo G, et al. Peripheral versus central potencies of N-type voltage-sensitive calcium channel blockers. Naunyn Schmiedebergs Arch Pharmacol. 1998;357:159-168.
Winquist RJ, Pan JQ, Gribkoff VK. Use-dependent blockade of Cav2.2 voltage-gated calcium channels for neuropathic pain. Biochem Pharmacol. 2005;70:489-499.
Woolf CJ, Mannion RJ. Neuropathic pain: aetiology, symptoms, mechanisms, and management. Lancet. 1999;353:1959-1964.
Woolf CJ, Salter MW. Neuronal plasticity: increasing the gain in pain. Science. 2000;288:1765-1769.
Woolfe G, MacDonald A. The evaluation of the analgesic action of pethidine hydrochloride. J Pharmacol Exp Ther. 1944;80:300-307.
Yaksh TL. Calcium channels as therapeutic targets in neuropathic pain. J Pain. 2006;7:S13-S30.
Zimmermann M. Ethical guidelines for investigations of experimental pain in conscious animals. Pain. 1983;16:109-110.