Examining dopamine D3 receptor occupancy by antipsychotic drugs via [3H]7-OH-DPAT ex vivo autoradiography and its cross-validation via c-fos immunohistochemistry in the rat brain

Eur J Pharmacol. 2014 Oct 5:740:669-75. doi: 10.1016/j.ejphar.2014.06.011. Epub 2014 Jun 23.

Abstract

Dopamine D3 receptors are a major target for drug discovery programs related to psychiatric disorders such as schizophrenia. The ability of a compound to occupy significant levels of D3 receptors is important for achieving therapeutic efficacy in both pre-clinical and clinical settings. Here we attempt to characterise antipsychotic drug-effects at D3 receptors by measuring receptor occupancy via ex-vivo [3H]7-OH-DPAT autoradiography, and further validating this outcome via analysis of Fos-like immunoreactivity (Fos-LI) in the rat major islands of Calleja (ICjM), a brain structure with high D3 expression. Rats were treated subcutaneously with haloperidol (0.04 mg/kg), clozapine (20 mg/kg) and olanzapine (0.63 mg/kg), the selective D2 antagonist L-741626 (2.5 mg/kg) and the selective D3 antagonist SB-277011-A (10 mg/kg). Doses were based on levels of D2 occupancy considered clinically relevant (60-80%). When measuring D3 occupancy, clozapine and SB-277011-A displayed meaningful levels of occupancy (60% and 77%, respectively), haloperidol and olanzapine showed limited occupancy (16% and 27%, respectively), whereas L-741626 showed no occupancy. There were no significant changes in ICjM Fos-LI after L-741626 and haloperidol treatment, minor but significant increases after olanzapine treatment, whereas highly significant increases were seen with SB-277011-A and clozapine. Additionally, pre-treating clozapine with the D1 antagonist SCH23390 caused a significant, albeit non-complete, reduction in Fos-LI, highlighting the D1 agonist property of clozapine. In conclusion, it appears that drugs occupying >50% D3 receptors produce robust increases in ICjM Fos-LI. This study may help to identify the appropriate D3 receptor antagonists that have the potential to be tested in the clinic.

Keywords: D(2); D(3); Major islands of Calleja; Receptor occupancy; c-fos.

MeSH terms

  • Animals
  • Antipsychotic Agents / pharmacology*
  • Autoradiography
  • Benzodiazepines / pharmacology
  • Clozapine / pharmacology
  • Dopamine Antagonists / pharmacology
  • Haloperidol / pharmacology
  • Indoles / pharmacology
  • Islands of Calleja / diagnostic imaging
  • Islands of Calleja / metabolism*
  • Male
  • Nitriles / pharmacology
  • Olanzapine
  • Piperidines / pharmacology
  • Proto-Oncogene Proteins c-fos / metabolism
  • Radiography
  • Rats, Wistar
  • Receptors, Dopamine D2 / metabolism*
  • Receptors, Dopamine D3 / antagonists & inhibitors
  • Receptors, Dopamine D3 / metabolism*
  • Tetrahydroisoquinolines / pharmacology
  • Tetrahydronaphthalenes

Substances

  • Antipsychotic Agents
  • Dopamine Antagonists
  • Indoles
  • Nitriles
  • Piperidines
  • Proto-Oncogene Proteins c-fos
  • Receptors, Dopamine D2
  • Receptors, Dopamine D3
  • SB 277011
  • Tetrahydroisoquinolines
  • Tetrahydronaphthalenes
  • Benzodiazepines
  • 3-(4-(4-chlorophenyl-4-hydroxypiperidino)methyl)indole
  • Clozapine
  • Haloperidol
  • Olanzapine
  • 7-hydroxy-2-N,N-dipropylaminotetralin