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 IC