Physiologically significant proteins often contain zinc, e.g., as a cofactor of enzymes or in the zinc finger motifs of transcription factors. The function of these zinc proteins can be modulated by ejecting zinc, as demonstrated by Rice et al.'s report of aromatic C-nitroso compounds that inhibit HIV-1 infectivity by ejecting zinc from zinc finger proteins. We report an alternate approach using novel zinc chelators to abstract zinc from HIV-EP1 (a human immunodeficiency virus type 1 enhancer binding protein with two C2H2-type zinc fingers that binds to the NF-κB recognition sequence) and thereby inhibit its DNA binding. We modified our previously reported compound Me2N-HPH (1) to prepare trityl derivative 2 and carboxyl derivatives 3-4. Competitive 1H NMR experiments showed that introducing a trityl group reduced zinc affinity (compound 2), while changing methyl ester to carboxyl groups enhanced it (compound 4 exhibited the highest zinc affinity). Electrophoretic mobility shift assays revealed that compounds 1-4 were more potent inhibitors of HIV-EP1 DNA binding than EDTA, even though EDTA has stronger zinc affinity—this superiority is likely due to the nitrogen-containing heterocyclic structure's favorable interactions (amino acid, hydrophobic, or electronic) that promote ternary complex formation with HIV-EP1-Zn. Zinc addition during or after the inhibition reaction fully restored HIV-EP1-DNA complex formation, confirming that inhibition resulted from zinc removal from HIV-EP1's zinc finger moiety (not competition for DNA binding, as verified by ethidium displacement and footprinting experiments). Thus, we developed novel zinc-binding heterocycles that effectively inhibit the DNA binding of the zinc finger protein HIV-EP1, providing a basis for the control and elucidation of various biochemical processes.