14 alpha,14' beta-[Dithiobis[(2-oxo-2,1-ethanediyl)imino]]bis (7,8-dihydromorphinone) and 14 alpha,14' beta-[dithiobis[(2-oxo-2,1- ethanediyl)imino]]bis[7,8-dihydro-N-(cyclopropylmethyl)normorphinone]: chemistry and opioid binding properties.
14 alpha,14' beta-[Dithiobis[(2-oxo-2,1-ethanediyl)imino]] bis(7,8-dihydromorphinone) (TAMO) (13) was synthesized by condensing 14 beta-amino-7,8-dihydromorphine (4) with acetylthioglycolyl chloride and hydrolyzing the resulting ester with mild base to give a mixture of the thiol 9 and the disulfide 13. Chromatography of the mixture resulted in conversion of the bulk of the thiol 9 to the disulfide 13 by air oxidation. The disulfide 13 was also prepared by condensing the tert-butyldimethylsilyl ether of 4 with the dithiodiglycolyl chloride and treating the resulting product with F- to give the desired product. The pure thiol 9 free of contamination with the disulfide was prepared by treating 13 with excess N-acetyl-L-cysteine and processing the reaction mixture without resorting to chromatography for purification. The corresponding N-(cyclopropylmethyl) nor compound 15 was prepared from the silyl ether 6 and acetylthioglycolyl chloride followed by hydrolysis, treatment with F-, and air oxidation. Incubation of bovine striatal membranes with 13 and 15 resulted in wash-resistant inhibition of the binding of the mu-selective peptide [3H][D-Ala2,(Me)Phe4,Gly(ol)5]-enkephalin (DAMGO). Incubation of membranes with mu but not kappa or delta ligands protected the mu binding sites from alkylation by 13 and 15. The wash-resistant inhibition of mu opioid binding was partially reversed by the addition of the reducing reagent dithiothreitol (DTT). A Scatchard plot of the effect of 13 and 15 on [3H]DAMGO binding showed that these affinity ligands caused a marked decrease in the Bmax value without affecting the Kd value. The wash-resistant inhibition of binding, the reduction in the number of binding sites, the partial reversal of wash-resistant inhibition of binding by DTT, and previously observed long-term antagonism of mu opioid receptors in vivo support the conclusion that 13 and 15 bind covalently to the mu opioid receptor.
Archer
S
S
Department of Chemistry, Cogswell Laboratory, Rensselaer Polytechnic Institute, Troy, New York 12180-3590.
Seyed-Mozaffari
A
A
Jiang
Q
Q
Bidlack
J M
JM
D000478:Alkylation; D000700:Analgesics / Q000138:chemical synthesis* / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D044482:Benzeneacetamides; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D003342:Corpus Striatum / Q000378:metabolism; D004220:Disulfides / Q000138:chemical synthesis* / Q000378:metabolism / Q000494:pharmacology; D004229:Dithiothreitol / Q000494:pharmacology; D004723:Endorphins / Q000378:metabolism; D020875:Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; D020881:Enkephalin, D-Penicillamine (2,5)-; D004745:Enkephalins / Q000378:metabolism; D004091:Hydromorphone / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000378:metabolism / Q000494:pharmacology; D007700:Kinetics; D015394:Molecular Structure; D011759:Pyrrolidines / Q000378:metabolism; D017473:Receptors, Opioid, kappa / Q000378:metabolism; D017450:Receptors, Opioid, mu / Q000378:metabolism; D014108:Tosylphenylalanyl Chloromethyl Ketone / Q000494:pharmacology
D000700:Analgesics; D044482:Benzeneacetamides; D004220:Disulfides; D004723:Endorphins; D004745:Enkephalins; D011759:Pyrrolidines; D017473:Receptors, Opioid, kappa; D017450:Receptors, Opioid, mu; D020875:Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; C076027:14-thioglycolamido-7,8-dihydromorphinone; C087555:14,14'-(dithiobis((2-oxo-2,1-ethanediyl)imino))bis(7,8-dihydro-N-(cyclopropylmethyl)normorphinone); D014108:Tosylphenylalanyl Chloromethyl Ketone; D020881:Enkephalin, D-Penicillamine (2,5)-; C045444:U 69593; D004091:Hydromorphone; D004229:Dithiothreitol