title: Design and cellular kinetics of dansyl-labeled CADA derivatives with anti-HIV and CD4 receptor down-modulating activity.
abstract: A new class of anti-retrovirals, cyclotriazadisulfonamide (CADA) and its derivatives, specifically down-regulate CD4, the main receptor of HIV, and prevent HIV infection in vitro. In this work, several CADA derivatives, chemically labeled with a fluorescent dansyl group, were evaluated for their biological features and cellular uptake kinetics. We identified a derivative KKD-016 with antiviral and CD4 down-modulating capabilities similar to those of the parental compound CADA. By using flow cytometry, we demonstrated that the dose-dependent cellular uptake of this derivative correlated with CD4 down-modulation. The uptake and activity of the dansyl-labeled compounds were not dependent on the level of expression of CD4 at the cell surface. Removal of the CADA compounds from the cell culture medium resulted in their release from the cells followed by a complete restoration of CD4 expression. The inability of several fluorescent CADA derivatives to down-modulate CD4 was not associated with their lower cellular uptake and was not reversed by facilitating their cell penetration by a surfactant. These results prove the successful integration of the dansyl fluorophore into the chemical structure of a CD4 down-modulating anti-HIV compound, and show the feasibility of tracking a receptor and its down-modulator simultaneously. These fluorescent CADA analogs with reversible CD4 down-regulating potency can now be applied in further studies on receptor modulation, and in the exploration of their potentials as preventive and therapeutic anti-HIV drugs.
authors.affiliation: Rega Institute for Medical Research, Department of Microbiology and Immunology, Katholieke Universiteit Leuven, Leuven, Belgium. kurt.vermeire@rega.kuleuven.be
authors.forename: Kurt
authors.initials: K
authors.lastname: Vermeire
authors.forename: Andrea
authors.initials: A
authors.lastname: Lisco
authors.forename: Jean-Charles
authors.initials: JC
authors.lastname: Grivel
authors.forename: Emily
authors.initials: E
authors.lastname: Scarbrough
authors.forename: Kaka
authors.initials: K
authors.lastname: Dey
authors.forename: Noah
authors.initials: N
authors.lastname: Duffy
authors.forename: Leonid
authors.initials: L
authors.lastname: Margolis
authors.forename: Thomas W
authors.initials: TW
authors.lastname: Bell
authors.forename: Dominique
authors.initials: D
authors.lastname: Schols
mesh_terms: D019380:Anti-HIV Agents / Q000138:chemical synthesis / Q000378:metabolism / Q000494:pharmacology*; D015704:CD4 Antigens / Q000235:genetics / Q000378:metabolism*; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D002460:Cell Line; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D003619:Dansyl Compounds / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation / Q000187:drug effects; D015195:Drug Design; D005434:Flow Cytometry; D015497:HIV-1 / Q000187:drug effects / Q000254:growth & development; D006571:Heterocyclic Compounds / Q000138:chemical synthesis / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D007150:Immunohistochemistry; D007700:Kinetics; D015394:Molecular Structure; D020442:Poloxamer / Q000737:chemistry; D012997:Solvents / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013449:Sulfonamides / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology; D013501:Surface-Active Agents / Q000737:chemistry; D014162:Transfection
chemical_list: C498074:9-benzyl-3-methylene-1,5-di-p-toluenesulfonyl-1,5,9-triazacyclododeane; D019380:Anti-HIV Agents; D015704:CD4 Antigens; D003619:Dansyl Compounds; D006571:Heterocyclic Compounds; C523232:KKD-015; C523233:KKD-016; D012997:Solvents; D013449:Sulfonamides; D013501:Surface-Active Agents; D020442:Poloxamer; D004121:Dimethyl Sulfoxide