Novel azido and isothiocyanato analogues of [3-(4-phenylalkylpiperazin-1-yl)propyl]bis(4-fluorophenyl)amines as potential irreversible ligands for the dopamine transporter.
Potential irreversible ligands were prepared, based on a series of 3-(1-piperazinyl)propyl-N,N-bis(4-fluorophenyl)amines, as molecular probes for the dopamine transporter (DAT). Both azido- and isothiocyanato-substituted phenylalkyl analogues were synthesized and evaluated for displacement of [(3)H]WIN 35 428 in rat caudate putamen tissue. All of the analogues showed moderate binding potencies at the DAT. The azido analogue, 16b, was radioiodinated and used to photolabel human DAT-transfected HEK 293 cell membranes. [(125)I]16b irreversibly labeled an approximately 80 kDa band corresponding to the DAT detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This radioligand provides a novel addition to the growing arsenal of structurally diverse irreversible ligands that are being used to identify binding domains on the DAT. Characterizing points of attachment of these irreversible probes to the DAT protein will ultimately help elucidate the three-dimensional arrangement of the transmembrane domains, identify individual binding sites of the DAT inhibitors, and direct future drug design.
Cao
Jianjing
J
Medicinal Chemistry and Psychobiology Sections, Intramural Research Program, National Institute on Drug Abuse, 5500 Nathan Shock Drive, Baltimore, Maryland 21224, USA.
Lever
John R
JR
Kopajtic
Theresa
T
Katz
Jonathan L
JL
Pham
Anh T
AT
Holmes
Muhsinah L
ML
Justice
Joseph B
JB
Newman
Amy Hauck
AH
D000818:Animals; D001386:Azides / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D001667:Binding, Competitive; D002460:Cell Line; D004159:Diphenylamine / Q000031:analogs & derivatives* / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D004298:Dopamine / Q000378:metabolism*; D050483:Dopamine Plasma Membrane Transport Proteins; D006801:Humans; D066298:In Vitro Techniques; D007457:Iodine Radioisotopes; D017879:Isothiocyanates / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D008024:Ligands; D008297:Male; D008562:Membrane Glycoproteins / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D019920:Photoaffinity Labels / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D010879:Piperazines / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D011699:Putamen / Q000378:metabolism; D011869:Radioligand Assay; D051381:Rats; D017207:Rats, Sprague-Dawley; D013329:Structure-Activity Relationship
C494716:3-(4-(3-iodo-4-azidophenylethyl)-1-piperazinyl)propyl-N,N-bis-(4-fluorophenyl)amine; D001386:Azides; D050483:Dopamine Plasma Membrane Transport Proteins; D007457:Iodine Radioisotopes; D017879:Isothiocyanates; D008024:Ligands; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D009419:Nerve Tissue Proteins; D019920:Photoaffinity Labels; D010879:Piperazines; C493214:SLC6A3 protein, human; C493216:Slc6a3 protein, rat; D004159:Diphenylamine; D004298:Dopamine