Studies of the biogenic amine transporters. VIII: identification of a novel partial inhibitor of dopamine uptake and dopamine transporter binding.
Using [125I]RTI-55 to label the dopamine transporter (DAT), our laboratory has consistently detected one binding site as well as one component of [3H]DA uptake. We report here the identification of a novel partial inhibitor of [3H]DA uptake and DAT binding (SoRI-9804). [125I]RTI-55 binding to the DAT (mouse caudate, rat caudate, HEK cells expressing the cloned DAT), the 5-HT transporter (rat brain), and [3H]DA uptake (rat caudate synaptosomes) were conducted using published procedures. 4-[(Diphenylmethyl)amino]-2-phenylquinazoline (SoRI-9804) was essentially inactive at SERT binding and resolved two DAT binding components in all three tissues, having high affinity (mean Ki of 465 nM) for about 40% of the binding sites and an essentially immeasurable Ki (> 100 microM) for the remaining 60% of the binding sites. The [3H]DA uptake experiments indicated that about 50% of uptake was SoRI-9804-sensitive. Saturation binding experiments showed that SoRI-9804 competitively inhibited [125I]RTI-55 binding to the SoRI-9804-sensitive binding component. To determine if the two binding sites discriminated by SoRI-9804 were regulated by the MAP kinase pathway, rat caudate synaptosomes were incubated in the absence or presence of 10 microM of PD98059, which inhibits activation of the MAP kinase pathway. The results indicated that inhibition of MAPK/ERK kinase decreased the total B(max) of the DAT by 90%. Treatment with PD98059 increased the proportion of the SoRI-9804-sensitive binding component from 68-80% of the total B(max). The PD98059 experiments suggest that inhibition of MAP kinase cannot explain the differential interaction of SoRI-9804 with the DAT. Viewed collectively, the present results indicate that SoRI-9804 discriminates two components of the DA transporter. Further studies will be needed to determine the underlying mechanism of this effect and if partial inhibition of DA uptake results in any unique behavioral effects.
Rothman
Richard B
RB
Clinical Psychopharmacology Section, Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, Baltimore, Maryland 21224, USA. rrothman@irp.nida.nih.gov
Dersch
Christina M
CM
Carroll
F Ivy
FI
Ananthan
Subramaniam
S
D000818:Animals; D001667:Binding, Competitive / Q000187:drug effects* / Q000502:physiology; D002352:Carrier Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D002421:Caudate Nucleus / Q000187:drug effects* / Q000378:metabolism; D003042:Cocaine / Q000031:analogs & derivatives / Q000037:antagonists & inhibitors / Q000494:pharmacology; D004298:Dopamine / Q000378:metabolism*; D050483:Dopamine Plasma Membrane Transport Proteins; D018765:Dopamine Uptake Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005419:Flavonoids / Q000494:pharmacology; D020935:MAP Kinase Signaling System / Q000187:drug effects / Q000502:physiology; D008297:Male; D008562:Membrane Glycoproteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D049990:Membrane Transport Modulators; D026901:Membrane Transport Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D009419:Nerve Tissue Proteins; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D011799:Quinazolines / Q000737:chemistry / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D050486:Serotonin Plasma Membrane Transport Proteins; D013574:Synaptosomes / Q000187:drug effects / Q000378:metabolism
C451939:4-((Diphenylmethyl)amino)-2-phenylquinazoline; D002352:Carrier Proteins; D050483:Dopamine Plasma Membrane Transport Proteins; D018765:Dopamine Uptake Inhibitors; D004791:Enzyme Inhibitors; D005419:Flavonoids; D008562:Membrane Glycoproteins; D049990:Membrane Transport Modulators; D026901:Membrane Transport Proteins; D009419:Nerve Tissue Proteins; D011799:Quinazolines; D050486:Serotonin Plasma Membrane Transport Proteins; C493216:Slc6a3 protein, rat; C493232:Slc6a4 protein, rat; C069723:2beta-carbomethoxy-3beta-(4-iodophenyl)tropane; D003042:Cocaine; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one; D004298:Dopamine