Small molecule inhibitors of PSD95-nNOS protein-protein interactions as novel analgesics

Wan Hung Lee, Zhili Xu, Nicole M. Ashpole, Andy Hudmon, Pushkar M. Kulkarni, Ganesh A. Thakur, Yvonne Y. Lai, Andrea G. Hohmann

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Abstract Aberrant increases in NMDA receptor (NMDAR) signaling contributes to central nervous system sensitization and chronic pain by activating neuronal nitric oxide synthase (nNOS) and generating nitric oxide (NO). Because the scaffolding protein postsynaptic density 95kDA (PSD95) tethers nNOS to NMDARs, the PSD95-nNOS complex represents a therapeutic target. Small molecule inhibitors IC87201 (EC5O: 23.94 μM) and ZL006 (EC50: 12.88 μM) directly inhibited binding of purified PSD95 and nNOS proteins in AlphaScreen without altering binding of PSD95 to ErbB4. Both PSD95-nNOS inhibitors suppressed glutamate-induced cell death with efficacy comparable to MK-801. IC87201 and ZL006 preferentially suppressed phase 2A pain behavior in the formalin test and suppressed allodynia induced by intraplantar complete Freund's adjuvant administration. IC87201 and ZL006 suppressed mechanical and cold allodynia induced by the chemotherapeutic agent paclitaxel (ED50s: 2.47 and 0.93 mg/kg i.p. for IC87201 and ZL006, respectively). Efficacy of PSD95-nNOS disruptors was similar to MK-801. Motor ataxic effects were induced by MK-801 but not by ZL006 or IC87201. Finally, MK-801 produced hyperalgesia in the tail-flick test whereas IC87201 and ZL006 did not alter basal nociceptive thresholds. Our studies establish the utility of using AlphaScreen and purified protein pairs to establish and quantify disruption of protein-protein interactions. Our results demonstrate previously unrecognized antinociceptive efficacy of ZL006 and establish, using two small molecules, a broad application for PSD95-nNOS inhibitors in treating neuropathic and inflammatory pain. Collectively, our results demonstrate that disrupting PSD95-nNOS protein-protein interactions is effective in attenuating pathological pain without producing unwanted side effects (i.e. motor ataxia) associated with NMDAR antagonists.

Original languageEnglish (US)
Article number5884
Pages (from-to)464-475
Number of pages12
JournalNeuropharmacology
Volume97
DOIs
StatePublished - 2015

Fingerprint

Post-Synaptic Density
Nitric Oxide Synthase Type I
Analgesics
Dizocilpine Maleate
Hyperalgesia
Proteins
N-Methyl-D-Aspartate Receptors
Central Nervous System Sensitization
Pain
Freund's Adjuvant
Neuralgia
Pain Measurement
Ataxia
Paclitaxel
Chronic Pain
2-((1H-benzo(d)(1,2,3)triazol-5-ylamino)methyl)-4,6-dichlorophenol
Glutamic Acid
Nitric Oxide
Cell Death

Keywords

  • Allodynia
  • Central sensitization
  • Complete Freund's adjuvant
  • motor ataxia
  • Neuronal nitric oxide synthase
  • Neuropathic pain
  • NMDAR
  • paclitaxel
  • Postsynaptic density 95

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience
  • Pharmacology

Cite this

Lee, W. H., Xu, Z., Ashpole, N. M., Hudmon, A., Kulkarni, P. M., Thakur, G. A., ... Hohmann, A. G. (2015). Small molecule inhibitors of PSD95-nNOS protein-protein interactions as novel analgesics. Neuropharmacology, 97, 464-475. [5884]. https://doi.org/10.1016/j.neuropharm.2015.05.038

Small molecule inhibitors of PSD95-nNOS protein-protein interactions as novel analgesics. / Lee, Wan Hung; Xu, Zhili; Ashpole, Nicole M.; Hudmon, Andy; Kulkarni, Pushkar M.; Thakur, Ganesh A.; Lai, Yvonne Y.; Hohmann, Andrea G.

In: Neuropharmacology, Vol. 97, 5884, 2015, p. 464-475.

Research output: Contribution to journalArticle

Lee, WH, Xu, Z, Ashpole, NM, Hudmon, A, Kulkarni, PM, Thakur, GA, Lai, YY & Hohmann, AG 2015, 'Small molecule inhibitors of PSD95-nNOS protein-protein interactions as novel analgesics', Neuropharmacology, vol. 97, 5884, pp. 464-475. https://doi.org/10.1016/j.neuropharm.2015.05.038
Lee, Wan Hung ; Xu, Zhili ; Ashpole, Nicole M. ; Hudmon, Andy ; Kulkarni, Pushkar M. ; Thakur, Ganesh A. ; Lai, Yvonne Y. ; Hohmann, Andrea G. / Small molecule inhibitors of PSD95-nNOS protein-protein interactions as novel analgesics. In: Neuropharmacology. 2015 ; Vol. 97. pp. 464-475.
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AU - Ashpole, Nicole M.

AU - Hudmon, Andy

AU - Kulkarni, Pushkar M.

AU - Thakur, Ganesh A.

AU - Lai, Yvonne Y.

AU - Hohmann, Andrea G.

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N2 - Abstract Aberrant increases in NMDA receptor (NMDAR) signaling contributes to central nervous system sensitization and chronic pain by activating neuronal nitric oxide synthase (nNOS) and generating nitric oxide (NO). Because the scaffolding protein postsynaptic density 95kDA (PSD95) tethers nNOS to NMDARs, the PSD95-nNOS complex represents a therapeutic target. Small molecule inhibitors IC87201 (EC5O: 23.94 μM) and ZL006 (EC50: 12.88 μM) directly inhibited binding of purified PSD95 and nNOS proteins in AlphaScreen without altering binding of PSD95 to ErbB4. Both PSD95-nNOS inhibitors suppressed glutamate-induced cell death with efficacy comparable to MK-801. IC87201 and ZL006 preferentially suppressed phase 2A pain behavior in the formalin test and suppressed allodynia induced by intraplantar complete Freund's adjuvant administration. IC87201 and ZL006 suppressed mechanical and cold allodynia induced by the chemotherapeutic agent paclitaxel (ED50s: 2.47 and 0.93 mg/kg i.p. for IC87201 and ZL006, respectively). Efficacy of PSD95-nNOS disruptors was similar to MK-801. Motor ataxic effects were induced by MK-801 but not by ZL006 or IC87201. Finally, MK-801 produced hyperalgesia in the tail-flick test whereas IC87201 and ZL006 did not alter basal nociceptive thresholds. Our studies establish the utility of using AlphaScreen and purified protein pairs to establish and quantify disruption of protein-protein interactions. Our results demonstrate previously unrecognized antinociceptive efficacy of ZL006 and establish, using two small molecules, a broad application for PSD95-nNOS inhibitors in treating neuropathic and inflammatory pain. Collectively, our results demonstrate that disrupting PSD95-nNOS protein-protein interactions is effective in attenuating pathological pain without producing unwanted side effects (i.e. motor ataxia) associated with NMDAR antagonists.

AB - Abstract Aberrant increases in NMDA receptor (NMDAR) signaling contributes to central nervous system sensitization and chronic pain by activating neuronal nitric oxide synthase (nNOS) and generating nitric oxide (NO). Because the scaffolding protein postsynaptic density 95kDA (PSD95) tethers nNOS to NMDARs, the PSD95-nNOS complex represents a therapeutic target. Small molecule inhibitors IC87201 (EC5O: 23.94 μM) and ZL006 (EC50: 12.88 μM) directly inhibited binding of purified PSD95 and nNOS proteins in AlphaScreen without altering binding of PSD95 to ErbB4. Both PSD95-nNOS inhibitors suppressed glutamate-induced cell death with efficacy comparable to MK-801. IC87201 and ZL006 preferentially suppressed phase 2A pain behavior in the formalin test and suppressed allodynia induced by intraplantar complete Freund's adjuvant administration. IC87201 and ZL006 suppressed mechanical and cold allodynia induced by the chemotherapeutic agent paclitaxel (ED50s: 2.47 and 0.93 mg/kg i.p. for IC87201 and ZL006, respectively). Efficacy of PSD95-nNOS disruptors was similar to MK-801. Motor ataxic effects were induced by MK-801 but not by ZL006 or IC87201. Finally, MK-801 produced hyperalgesia in the tail-flick test whereas IC87201 and ZL006 did not alter basal nociceptive thresholds. Our studies establish the utility of using AlphaScreen and purified protein pairs to establish and quantify disruption of protein-protein interactions. Our results demonstrate previously unrecognized antinociceptive efficacy of ZL006 and establish, using two small molecules, a broad application for PSD95-nNOS inhibitors in treating neuropathic and inflammatory pain. Collectively, our results demonstrate that disrupting PSD95-nNOS protein-protein interactions is effective in attenuating pathological pain without producing unwanted side effects (i.e. motor ataxia) associated with NMDAR antagonists.

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KW - Postsynaptic density 95

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