title: Structure-based identification and neutralization mechanism of tyrosine sulfate mimetics that inhibit HIV-1 entry.
abstract: Tyrosine sulfate-mediated interactions play an important role in HIV-1 entry. After engaging the CD4 receptor at the cell surface, the HIV-1 gp120 glycoprotein binds to the CCR5 co-receptor via an interaction that requires two tyrosine sulfates, at positions 10 and 14 in the CCR5-N terminus. Building on previous structure determinations of this interaction, here we report the targeting of these tyrosine sulfate binding sites for drug design through in silico screening of small molecule libraries, identification of lead compounds, and characterization of biological activity. A class of tyrosine sulfate-mimicking small molecules containing a "phenyl sulfonate-linker-aromatic" motif was identified that specifically inhibited binding of gp120 to the CCR5-N terminus as well as to sulfated antibodies that recognize the co-receptor binding region on gp120. The most potent of these compounds bound gp120 with low micromolar affinity and its CD4-induced conformation with K(D)'s as tight as ∼50 nM. Neutralization experiments suggested the targeted site to be conformationally inaccessible prior to CD4 engagement. Primary HIV-1 isolates were weakly neutralized, preincubation with soluble CD4 enhanced neutralization, and engineered isolates with increased dependence on the N terminus of CCR5 or with reduced conformational barriers were neutralized with IC(50) values as low as ∼1 μM. These results reveal the potential of targeting the tyrosine sulfate interactions of HIV-1 and provide insight into how mechanistic barriers, evolved by HIV-1 to evade antibody recognition, also restrict small-molecule-mediated neutralization.
authors.affiliation: Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892, United States.
authors.forename: Priyamvada
authors.initials: P
authors.lastname: Acharya
authors.forename: Cajetan
authors.initials: C
authors.lastname: Dogo-Isonagie
authors.forename: Judith M
authors.initials: JM
authors.lastname: LaLonde
authors.forename: Son N
authors.initials: SN
authors.lastname: Lam
authors.forename: George J
authors.initials: GJ
authors.lastname: Leslie
authors.forename: Mark K
authors.initials: MK
authors.lastname: Louder
authors.forename: Leah L
authors.initials: LL
authors.lastname: Frye
authors.forename: Asim K
authors.initials: AK
authors.lastname: Debnath
authors.forename: Jeremy R
authors.initials: JR
authors.lastname: Greenwood
authors.forename: Timothy S
authors.initials: TS
authors.lastname: Luongo
authors.forename: Loïc
authors.initials: L
authors.lastname: Martin
authors.forename: K Shawn
authors.initials: KS
authors.lastname: Watts
authors.forename: James A
authors.initials: JA
authors.lastname: Hoxie
authors.forename: John R
authors.initials: JR
authors.lastname: Mascola
authors.forename: Carole A
authors.initials: CA
authors.lastname: Bewley
authors.forename: Peter D
authors.initials: PD
authors.lastname: Kwong
mesh_terms: D019380:Anti-HIV Agents / Q000737:chemistry* / Q000494:pharmacology*; D015704:CD4 Antigens / Q000276:immunology; D015699:HIV Envelope Protein gp120 / Q000276:immunology / Q000378:metabolism; D015658:HIV Infections / Q000188:drug therapy; D015497:HIV-1 / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D008958:Models, Molecular; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D053586:Virus Internalization / Q000187:drug effects*
chemical_list: D019380:Anti-HIV Agents; D015704:CD4 Antigens; D015699:HIV Envelope Protein gp120; C005662:tyrosine O-sulfate; D014443:Tyrosine