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Characterization of the Effects of L-4-Chlorokynurenine on Nociception in Rodents

      Highlights

      • The NMDA receptor (NMDAR) is regulated by the glycineB-site (GlyB).
      • 4-Cl-kynurenine (AV-101) is a prodrug that does not bind to the NMDAR GlyB.
      • AV-101 metabolite, 7-Cl-kynurenic acid, is a potent NMDAR GlyB antagonist.
      • Brain 7-Cl-kynurenic acid concentrations achieved exceed the IC50 of GlyB.
      • AV-101 reduces tissue/nerve injury pain with minimal adverse effects.

      Abstract

      Upon systemic administration in rats, the prodrug L-4-chlorokynurenine (4-Cl-KYN; AV-101; VistaGen Therapeutics, Inc, South San Francisco, CA) is rapidly absorbed, actively transported across the blood-brain barrier, and converted in astrocytes to 7-chlorokynurenic acid (7-Cl-KYNA), a potent and specific antagonist of the glycine B coagonist site of the N-methyl-D-aspartate (NMDA) receptor. We examined the effects of 4-Cl-KYN in several rat models of hyperalgesia and allodynia and determined the concentrations of 4-Cl-KYN and newly produced 7-Cl-KYNA in serum, brain, and spinal cord. Adult male rats were given 4-Cl-KYN (56, 167, 500 mg/kg), the NMDA receptor antagonist MK-801 (.1, .3, 1.0 mg/kg), or gabapentin (33, 100, 300 mg/kg) intraperitoneally, and were then examined on rotarod, intraplantar formalin-evoked flinching, thermal escape in the normal and carrageenan-inflamed paw, and allodynia after sciatic nerve ligation. Our conclusions show that after systemic delivery, the highest 2 doses (167 and 500 mg/kg) of 4-Cl-KYN yielded brain concentrations of 7-Cl-KYNA exceeding its half maximal inhibitory concentration (IC50) at the glycine B site and resulted in dose-dependent antihyperalgesia in the 4 models of facilitated processing associated with tissue inflammation and nerve injury. On the basis of the relative dose requirements for analgesic actions and side effect profiles from these experiments, 4-Cl-KYN is predicted to have antihyperalgesic efficacy and a therapeutic ratio equal to gabapentin and superior to MK-801.

      Perspective

      These studies show that systemic administration of the prodrug 4-Cl-KYN produces high central nervous system levels of 7-Cl-KYNA, a potent and highly selective antagonist of the NMDA receptor. Compared with other drugs tested, 4-Cl-KYN has robust antinociceptive effects with a better side effect profile, highlighting its potential for treating hyperpathic pain states.

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      References

        • Albers G.W.
        • Clark W.M.
        • Atkinson R.P.
        • Madden K.
        • Data J.L.
        • Whitehouse M.J.
        Dose escalation study of the NMDA glycine-site antagonist licostinel in acute ischemic stroke.
        Stroke. 1999; 30: 508-513
        • Beagles K.E.
        • Morrison P.F.
        • Heyes M.P.
        Quinolinic acid in vivo synthesis rates, extracellular concentrations, and intercompartmental distributions in normal and immune-activated brain as determined by multiple-isotope microdialysis.
        J Neurochem. 1998; 70: 281-291
        • Bonina F.P.
        • Arenare L.
        • Ippolito R.
        • Boatto G.
        • Battaglia G.
        • Bruno V.
        • de Caprariis P.
        Synthesis, pharmacokinetics and anticonvulsant activity of 7-chlorokynurenic acid prodrugs.
        Int J Pharm. 2000; 202: 79-88
        • Bradesi S.
        Role of spinal cord glia in the central processing of peripheral pain perception.
        Neurogastroenterol Motil. 2010; 22: 499-511
        • Brennan T.J.
        • Zahn P.K.
        Effect of intrathecal ACEA-1021 in a rat model for postoperative pain.
        J Pain. 2000; 1: 279-284
        • Chaplan S.R.
        • Bach F.W.
        • Pogrel J.W.
        • Chung J.M.
        • Yaksh T.L.
        Quantitative assessment of tactile allodynia in the rat paw.
        J Neurosci Methods. 1994; 53: 55-63
        • Chaplan S.R.
        • Malmberg A.B.
        • Yaksh T.L.
        Efficacy of spinal NMDA receptor antagonism in formalin hyperalgesia and nerve injury evoked allodynia in the rat.
        J Pharmacol Exp Ther. 1997; 280: 829-838
        • Chapman V.
        • Dickenson A.H.
        Time-related roles of excitatory amino acid receptors during persistent noxiously evoked responses of rat dorsal horn neurones.
        Brain Res. 1995; 703: 45-50
        • Chen H.S.
        • Lipton S.A.
        The chemical biology of clinically tolerated NMDA receptor antagonists.
        J Neurochem. 2006; 97: 1611-1626
        • Chiarugi A.
        • Moroni F.
        Quinolinic acid formation in immune-activated mice: Studies with (m-nitrobenzoyl)-alanine (mNBA) and 3,4-dimethoxy-[-N-4-(-3-nitrophenyl)thiazol-2yl]-benzenesul fonamide (Ro 61-8048), two potent and selective inhibitors of kynurenine hydroxylase.
        Neuropharmacology. 1999; 38: 1225-1233
        • Coan E.J.
        • Saywood W.
        • Collingridge G.L.
        MK-801 blocks NMDA receptor-mediated synaptic transmission and long term potentiation in rat hippocampal slices.
        Neurosci Lett. 1987; 80: 111-114
        • Coderre T.J.
        • Melzack R.
        The contribution of excitatory amino acids to central sensitization and persistent nociception after formalin-induced tissue injury.
        J Neurosci. 1992; 12: 3665-3670
        • Coderre T.J.
        • Van Empel I.
        The utility of excitatory amino acid (EAA) antagonists as analgesic agents. I. Comparison of the antinociceptive activity of various classes of EAA antagonists in mechanical, thermal and chemical nociceptive tests.
        Pain. 1994; 59: 345-352
        • Davies A.
        • Hendrich J.
        • Van Minh A.T.
        • Wratten J.
        • Douglas L.
        • Dolphin A.C.
        Functional biology of the alpha(2)delta subunits of voltage-gated calcium channels.
        Trends Pharmacol Sci. 2007; 28: 220-228
        • Davies S.N.
        • Lodge D.
        Evidence for involvement of N-methylaspartate receptors in ‘wind-up’ of class 2 neurones in the dorsal horn of the rat.
        Brain Res. 1987; 424: 402-406
        • Dickenson A.H.
        A cure for wind up: NMDA receptor antagonists as potential analgesics.
        Trends Pharmacol Sci. 1990; 11: 307-309
        • Dickenson A.H.
        • Aydar E.
        Antagonism at the glycine site on the NMDA receptor reduces spinal nociception in the rat.
        Neurosci Lett. 1991; 121: 263-266
        • Dickenson A.H.
        • Sullivan A.F.
        Evidence for a role of the NMDA receptor in the frequency dependent potentiation of deep rat dorsal horn nociceptive neurones following C fibre stimulation.
        Neuropharmacology. 1987; 26: 1235-1238
        • Dubner R.
        • Ruda M.A.
        Activity-dependent neuronal plasticity following tissue injury and inflammation.
        Trends Neurosci. 1992; 15: 96-103
        • Espey M.G.
        • Moffett J.R.
        • Namboodiri M.A.
        Temporal and spatial changes of quinolinic acid immunoreactivity in the immune system of lipopolysaccharide-stimulated mice.
        J Leukoc Biol. 1995; 57: 199-206
        • Guidetti P.
        • Wu H.Q.
        • Schwarcz R.
        In situ produced 7-chlorokynurenate provides protection against quinolinate- and malonate-induced neurotoxicity in the rat striatum.
        Exp Neurol. 2000; 163: 123-130
        • Haley J.E.
        • Dickenson A.H.
        Evidence for spinal N-methyl-d-aspartate receptor involvement in prolonged chemical nociception in the rat.
        Brain Res. 2016; 1645: 58-60
        • Haley J.E.
        • Sullivan A.F.
        • Dickenson A.H.
        Evidence for spinal N-methyl-D-aspartate receptor involvement in prolonged chemical nociception in the rat.
        Brain Res. 1990; 518: 218-226
        • Herrero J.F.
        • Laird J.M.
        • Lopez-Garcia J.A.
        Wind-up of spinal cord neurones and pain sensation: Much ado about something?.
        Prog Neurobiol. 2000; 61: 169-203
        • Heyes M.P.
        • Saito K.
        • Major E.O.
        • Milstien S.
        • Markey S.P.
        • Vickers J.H.
        A mechanism of quinolinic acid formation by brain in inflammatory neurological disease. Attenuation of synthesis from L-tryptophan by 6-chlorotryptophan and 4-chloro-3-hydroxyanthranilate.
        Brain. 1993; 116: 1425-1450
        • Hilmas C.
        • Pereira E.F.
        • Alkondon M.
        • Rassoulpour A.
        • Schwarcz R.
        • Albuquerque E.X.
        The brain metabolite kynurenic acid inhibits alpha7 nicotinic receptor activity and increases non-alpha7 nicotinic receptor expression: Physiopathological implications.
        J Neurosci. 2001; 21: 7463-7473
        • Hokari M.
        • Wu H.Q.
        • Schwarcz R.
        • Smith Q.R.
        Facilitated brain uptake of 4-chlorokynurenine and conversion to 7-chlorokynurenic acid.
        Neuroreport. 1996; 8: 15-18
        • Kemp J.A.
        • Foster A.C.
        • Leeson P.D.
        • Priestley T.
        • Tridgett R.
        • Iversen L.L.
        • Woodruff G.N.
        7-Chlorokynurenic acid is a selective antagonist at the glycine modulatory site of the N-methyl-D-aspartate receptor complex.
        Proc Natl Acad Sci U S A. 1988; 85: 6547-6550
        • Kim S.H.
        • Chung J.M.
        An experimental model for peripheral neuropathy produced by segmental spinal nerve ligation in the rat.
        Pain. 1992; 50: 355-363
        • Kiss C.
        • Ceresoli-Borroni G.
        • Guidetti P.
        • Zielke C.L.
        • Zielke H.R.
        • Schwarcz R.
        Kynurenate production by cultured human astrocytes.
        J Neural Transm (Vienna). 2003; 110: 1-14
        • Kita T.
        • Morrison P.F.
        • Heyes M.P.
        • Markey S.P.
        Effects of systemic and central nervous system localized inflammation on the contributions of metabolic precursors to the L-kynurenine and quinolinic acid pools in brain.
        J Neurochem. 2002; 82: 258-268
        • Kolhekar R.
        • Meller S.T.
        • Gebhart G.F.
        N-methyl-D-aspartate receptor-mediated changes in thermal nociception: Allosteric modulation at glycine and polyamine recognition sites.
        Neuroscience. 1994; 63: 925-936
        • Laird J.M.
        • Mason G.S.
        • Webb J.
        • Hill R.G.
        • Hargreaves R.J.
        Effects of a partial agonist and a full antagonist acting at the glycine site of the NMDA receptor on inflammation-induced mechanical hyperalgesia in rats.
        Br J Pharmacol. 1996; 117: 1487-1492
        • Lees K.R.
        • Asplund K.
        • Carolei A.
        • Davis S.M.
        • Diener H.C.
        • Kaste M.
        • Orgogozo J.M.
        • Whitehead J.
        Glycine antagonist (gavestinel) in neuroprotection (GAIN International) in patients with acute stroke: A randomised controlled trial. GAIN International Investigators.
        Lancet. 2000; 355: 1949-1954
        • Lee S.C.
        • Schwarcz R.
        Excitotoxic injury stimulates pro-drug-induced 7-chlorokynurenate formation in the rat striatum in vivo.
        Neurosci Lett. 2001; 304: 185-188
        • Liu B.B.
        • Luo L.
        • Liu X.L.
        • Geng D.
        • Liu Q.
        • Yi L.T.
        7-Chlorokynurenic acid (7-CTKA) produces rapid antidepressant-like effects: Through regulating hippocampal microRNA expressions involved in TrkB-ERK/Akt signaling pathways in mice exposed to chronic unpredictable mild stress.
        Psychopharmacology (Berl). 2015; 232: 541-550
        • Lutfy K.
        • Weber E.
        Attenuation of nociceptive responses by ACEA-1021, a competitive NMDA receptor/glycine site antagonist, in the mice.
        Brain Res. 1996; 743: 17-23
        • Mothet J.P.
        • Parent A.T.
        • Wolosker H.
        • Brady Jr., R.O.
        • Linden D.J.
        • Ferris C.D.
        • Rogawski M.A.
        • Snyder S.H.
        D-serine is an endogenous ligand for the glycine site of the N-methyl-D-aspartate receptor.
        Proc Natl Acad Sci U S A. 2000; 97: 4926-4931
        • Nishiyama T.
        • Yaksh T.L.
        • Weber E.
        Effects of intrathecal NMDA and non-NMDA antagonists on acute thermal nociception and their interaction with morphine.
        Anesthesiology. 1998; 89: 715-722
        • Okuno E.
        • Nakamura M.
        • Schwarcz R.
        Two kynurenine aminotransferases in human brain.
        Brain Res. 1991; 542: 307-312
        • Okuno E.
        • Schmidt W.
        • Parks D.A.
        • Nakamura M.
        • Schwarcz R.
        Measurement of rat brain kynurenine aminotransferase at physiological kynurenine concentrations.
        J Neurochem. 1991; 57: 533-540
        • Old E.A.
        • Clark A.K.
        • Malcangio M.
        The role of glia in the spinal cord in neuropathic and inflammatory pain.
        Handb Exp Pharmacol. 2015; 227: 145-170
        • Paoletti P.
        • Bellone C.
        • Zhou Q.
        NMDA receptor subunit diversity: Impact on receptor properties, synaptic plasticity and disease.
        Nat Rev Neurosci. 2013; 14: 383-400
        • Pukelsheim F.
        The Three Sigma Rule.
        Am Stat. 1994; 48: 88-91
        • Quartaroli M.
        • Carignani C.
        • Dal Forno G.
        • Mugnaini M.
        • Ugolini A.
        • Arban R.
        • Bettelini L.
        • Maraia G.
        • Belardetti F.
        • Reggiani A.
        • Trist D.G.
        • Ratti E.
        • Di Fabio R.
        • Corsi M.
        Potent antihyperalgesic activity without tolerance produced by glycine site antagonist of N-methyl-D-aspartate receptor GV196771A.
        J Pharmacol Exp Ther. 1999; 290: 158-169
        • Quartaroli M.
        • Fasdelli N.
        • Bettelini L.
        • Maraia G.
        • Corsi M.
        GV196771A, an NMDA receptor/glycine site antagonist, attenuates mechanical allodynia in neuropathic rats and reduces tolerance induced by morphine in mice.
        Eur J Pharmacol. 2001; 430: 219-227
        • Rao T.S.
        • Gray N.M.
        • Dappen M.S.
        • Cler J.A.
        • Mick S.J.
        • Emmett M.R.
        • Iyengar S.
        • Monahan J.B.
        • Cordi A.A.
        • Wood P.L.
        Indole-2-carboxylates, novel antagonists of the N-methyl-D-aspartate (NMDA)-associated glycine recognition sites: In vivo characterization.
        Neuropharmacology. 1993; 32: 139-147
        • Ren K.
        • Williams G.M.
        • Hylden J.L.
        • Ruda M.A.
        • Dubner R.
        The intrathecal administration of excitatory amino acid receptor antagonists selectively attenuated carrageenan-induced behavioral hyperalgesia in rats.
        Eur J Pharmacol. 1992; 219: 235-243
        • Sacco R.L.
        • DeRosa J.T.
        • Haley Jr., E.C.
        • Levin B.
        • Ordronneau P.
        • Phillips S.J.
        • Rundek T.
        • Snipes R.G.
        • Thompson J.L.
        Glycine antagonist in neuroprotection for patients with acute stroke: GAIN Americas: A randomized controlled trial.
        JAMA. 2001; 285: 1719-1728
        • Saito K.
        • Markey S.P.
        • Heyes M.P.
        6-Chloro-D,L-tryptophan, 4-chloro-3-hydroxyanthranilate and dexamethasone attenuate quinolinic acid accumulation in brain and blood following systemic immune activation.
        Neurosci Lett. 1994; 178: 211-215
        • Saito K.
        • Markey S.P.
        • Heyes M.P.
        Effects of immune activation on quinolinic acid and neuroactive kynurenines in the mouse.
        Neuroscience. 1992; 51: 25-39
        • Salituro F.G.
        • Tomlinson R.C.
        • Baron B.M.
        • Palfreyman M.G.
        • McDonald I.A.
        • Schmidt W.
        • Wu H.Q.
        • Guidetti P.
        • Schwarcz R.
        Enzyme-activated antagonists of the strychnine-insensitive glycine/NMDA receptor.
        J Med Chem. 1994; 37: 334-336
        • Schwarcz R.
        • Bruno J.P.
        • Muchowski P.J.
        • Wu H.Q.
        Kynurenines in the mammalian brain: When physiology meets pathology.
        Nat Rev Neurosci. 2012; 13: 465-477
        • Tan-No K.
        • Esashi A.
        • Nakagawasai O.
        • Niijima F.
        • Furuta S.
        • Sato T.
        • Satoh S.
        • Yasuhara H.
        • Tadano T.
        Intrathecally administered D-cycloserine produces nociceptive behavior through the activation of N-methyl-D-aspartate receptor ion-channel complex acting on the glycine recognition site.
        J Pharmacol Sci. 2007; 104: 39-45
        • Todd W.P.
        • Carpenter B.K.
        • Schwarcz R.
        Preparation of 4-halo-3-hydroxyanthranilates and demonstration of their inhibition of 3-hydroxyanthranilate oxygenase activity in rat and human brain tissue.
        Prep Biochem. 1989; 19: 155-165
        • Traynelis S.F.
        • Wollmuth L.P.
        • McBain C.J.
        • Menniti F.S.
        • Vance K.M.
        • Ogden K.K.
        • Hansen K.B.
        • Yuan H.
        • Myers S.J.
        • Dingledine R.
        Glutamate receptor ion channels: Structure, regulation, and function.
        Pharmacol Rev. 2010; 62: 405-496
      1. USDA: Animal Welfare, 9 CFR Part 3. Federal Register. Vol 32, United States Department of Agriculture, February 15, 6426-6505, 1991

        • Wallace M.S.
        • Rowbotham M.C.
        • Katz N.P.
        • Dworkin R.H.
        • Dotson R.M.
        • Galer B.S.
        • Rauck R.L.
        • Backonja M.M.
        • Quessy S.N.
        • Meisner P.D.
        A randomized, double-blind, placebo-controlled trial of a glycine antagonist in neuropathic pain.
        Neurology. 2002; 59: 1694-1700
      2. Wallace M, White A, Grako KA, Lane R, Cato A, Snodgrass HR: Randomized, double-blind, placebo controlled, dose-escalation study: investigation of the safety, pharmacokinetics, and antihyperalgesic activity of L 4 chlorokynurenine in healthy volunteers. Scand J Pain. Accepted, 2017

        • Wood P.L.
        • Mahmood S.A.
        • Moskal J.R.
        Antinociceptive action of GLYX-13: An N-methyl-D-aspartate receptor glycine site partial agonist.
        Neuroreport. 2008; 19: 1059-1061
        • Woolf C.J.
        Central sensitization: Implications for the diagnosis and treatment of pain.
        Pain. 2011; 152: S2-S15
        • Woolf C.J.
        • Thompson S.W.
        The induction and maintenance of central sensitization is dependent on N-methyl-D-aspartic acid receptor activation; implications for the treatment of post-injury pain hypersensitivity states.
        Pain. 1991; 44: 293-299
        • Wu H.Q.
        • Rassoulpour A.
        • Goodman J.H.
        • Scharfman H.E.
        • Bertram E.H.
        • Schwarcz R.
        Kynurenate and 7-chlorokynurenate formation in chronically epileptic rats.
        Epilepsia. 2005; 46: 1010-1016
        • Wu H.Q.
        • Salituro F.G.
        • Schwarcz R.
        Enzyme-catalyzed production of the neuroprotective NMDA receptor antagonist 7-chlorokynurenic acid in the rat brain in vivo.
        Eur J Pharmacol. 1997; 319: 13-20
        • Yaksh T.L.
        • Hua X.Y.
        • Kalcheva I.
        • Nozaki-Taguchi N.
        • Marsala M.
        The spinal biology in humans and animals of pain states generated by persistent small afferent input.
        Proc Natl Acad Sci U S A. 1999; 96: 7680-7686
        • Yaksh T.L.
        • Ozaki G.
        • McCumber D.
        • Rathbun M.
        • Svensson C.
        • Malkmus S.
        • Yaksh M.C.
        An automated flinch detecting system for use in the formalin nociceptive bioassay.
        J Appl Physiol (1985). 2001; 90: 2386-2402
        • Yamamoto T.
        • Yaksh T.L.
        Spinal pharmacology of thermal hyperesthesia induced by constriction injury of sciatic nerve. Excitatory amino acid antagonists.
        Pain. 1992; 49: 121-128
        • Yates J.R.
        • Heyes M.P.
        • Blight A.R.
        4-Chloro-3-hydroxyanthranilate reduces local quinolinic acid synthesis, improves functional recovery, and preserves white matter after spinal cord injury.
        J Neurotrauma. 2006; 23: 866-881
        • Zanos P.
        • Piantadosi S.C.
        • Wu H.Q.
        • Pribut H.J.
        • Dell M.J.
        • Can A.
        • Snodgrass H.R.
        • Zarate Jr., C.A.
        • Schwarcz R.
        • Gould T.D.
        The prodrug 4-chlorokynurenine causes ketamine-like antidepressant effects, but not side effects, by NMDA/glycineB-site inhibition.
        J Pharmacol Exp Ther. 2015; 355: 76-85
        • Zhou H.Y.
        • Chen S.R.
        • Pan H.L.
        Targeting N-methyl-D-aspartate receptors for treatment of neuropathic pain.
        Expert Rev Clin Pharmacol. 2011; 4: 379-388
        • Zhu W.L.
        • Wang S.J.
        • Liu M.M.
        • Shi H.S.
        • Zhang R.X.
        • Liu J.F.
        • Ding Z.B.
        • Lu L.
        Glycine site N-methyl-D-aspartate receptor antagonist 7-CTKA produces rapid antidepressant-like effects in male rats.
        J Psychiatry Neurosci. 2013; 38: 306-316