Identification of pyrimidine structure-based compounds as allosteric ligands of the dopamine transporter as therapeutic agents for NeuroHIV.

The disruption of dopamine (DA) neurotransmission by the HIV-1 transactivator of transcription (Tat) during HIV-1 infection has been linked to the development of neurocognitive disorders, even under combined antiretroviral therapy treatment. We have demonstrated that Southern Research Institute (SRI) 32742, a novel allosteric modulator of DA transporter (DAT), attenuates cocaine- and Tat-binding to DAT, alleviates Tat-induced cognitive deficits and potentiation of cocaine reward in inducible Tat transgenic mice. The current study determined the in vitro pharmacological profile of SRI-32743 and its optimized second-generation analogs and their effects as allosteric modulators. Through structure-activity relationship studies of SRI-32743, 170 compounds were synthesized and evaluated for their ability to modulate DAT function. We identified 21 analogs as atypical competitors of DAT (maximum attributable drug effect, ≤60%). Four compounds, SRI-46564, SRI-47056, SRI-46286, and SRI-47867, displayed IC50 values for [3H]DA uptake inhibition from 9.33 ± 0.50 to 0.96 ± 0.05 μM and from 3.96 ± 1.36 to 1.29 ± 0.19 for DAT binding, respectively. The 4 analogs also displayed high potency at 2 different concentrations (0.5 nM and 0.05 nM) to attenuate Tat-induced inhibition of [3H]DA uptake and cocaine-mediated dissociation of [3H]WIN35,428 binding in Chinese hamster ovary cells expressing human DAT, suggesting that the effects occur through an allosteric mechanism. In further ex vivo studies using fast scan cyclic voltammetry, we demonstrated that the analogs do not disrupt the baseline phasic-like DA release. These findings provide a new insight into the potential for development of novel therapeutic agents to attenuate DAT-Tat interactions to normalize DA neurotransmission in NeuroHIV. SIGNIFICANCE STATEMENT: The allosteric inhibition of the dopamine (DA) transporter by the HIV-1 transactivator of transcription (Tat) disrupts DA homeostasis, leading to HIV-associated neurocognitive disorders. Analogs of Southern Research Institute 32743, a novel allosteric modulator of the Tat-DA transporter (DAT) interaction, were evaluated in the current study and characterized as atypical ligands of DA uptake. Four novel lead compounds demonstrated high potency to attenuate Tat-induced inhibition of human DAT-mediated DA uptake in an allosteric modulatory manner with no effects on the dynamics of DA uptake-release in DAT.