title: Dipyridamole analogs as pharmacological inhibitors of equilibrative nucleoside transporters. Identification of novel potent and selective inhibitors of the adenosine transporter function of human equilibrative nucleoside transporter 4 (hENT4).
abstract: To identify needed human equilibrative nucleoside transporter 4 (hENT4) inhibitors, we cloned and stably expressed the recombinant protein in PK15NTD (nucleoside transporter deficient) cells, and, investigated its interaction with a series of dipyridamole analogs synthesized in our laboratory. Compounds were tested in this newly established hENT4 expressing system as well in previous stably expressed hENT1 and hENT2 expressing systems. Of the dipyridamole analogs evaluated, about one fourth of the compounds inhibited hENT4 with higher potencies than dipyridamole. The most potent of them, Compound 30 displayed an IC₅₀ of 74.4 nM, making it about 38 times more potent than dipyridamole (IC₅₀=2.8 μM), and selectivities of about 80-fold and 20-fold relative to ENT1 and ENT2, respectively. Structure-activity relationship showed nitrogen-containing monocyclic rings and noncyclic substituents at the 4- and 8-positions of the pyrimido[5,4-d]pyrimidine were important for the inhibitory activity against hENT4. The most potent and selective hENT4 inhibitors tended to have a 2,6-di(N-monohydroxyethyl) substitution on the pyrimidopyrimidine ring system. The inhibitors of hENT4 identified in this study are the most selective and potent inhibitors of hENT4 adenosine transporter function to date, and should serve as useful pharmacological/biochemical tools and/or potential leads for ENT4-based therapeutics. Also, the new hENT4-expressing PK15 cell line established will serve as a useful screening tool for the discovery and design of hENT4 ligands.
authors.affiliation: Department of Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health Science Center, Memphis, TN 38163, USA.
authors.forename: Chunmei
authors.initials: C
authors.lastname: Wang
authors.forename: Wenwei
authors.initials: W
authors.lastname: Lin
authors.forename: Hilaire
authors.initials: H
authors.lastname: Playa
authors.forename: Shan
authors.initials: S
authors.lastname: Sun
authors.forename: Keyuna
authors.initials: K
authors.lastname: Cameron
authors.forename: John K
authors.initials: JK
authors.lastname: Buolamwini
mesh_terms: D000818:Animals; D015153:Blotting, Western; D018929:Cell Culture Techniques; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004176:Dipyridamole / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology*; D015195:Drug Design; D033705:Equilibrative Nucleoside Transport Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics; D006801:Humans; D008024:Ligands; D015394:Molecular Structure; D033703:Nucleoside Transport Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics; D013329:Structure-Activity Relationship; D013552:Swine; D014162:Transfection
chemical_list: D033705:Equilibrative Nucleoside Transport Proteins; D008024:Ligands; D033703:Nucleoside Transport Proteins; C495988:SLC29A4 protein, human; C444719:adenosine transporter; D004176:Dipyridamole
keywords: Dipyridamole analogs; Equilibrative nucleoside transporters; Human equilibrative nucleoside transporter 4; Structure–activity relationships
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