The HIV-1 envelope glycoprotein gp120 is the key determinant for the anti-HIV-1 entry activity of IC9564. A T198P mutation in the gp120 of the HIV-1 primary isolate, DH012, drastically increases IC9564 sensitivity, which can be reversed by growing the virus in the presence of IC9564. The reversed resistant variants contain a P198S mutation that fully confers the drug-resistant phenotype. Although the amino acid residue at position 198 of gp120 can alter IC9564 sensitivity, results from this study suggest that T198 is not the direct target of the compound. The mutation at position 198 appears to affect the conformation of gp120 and subsequently decreases the accessibility of the drug target. This conformational effect is evidenced by the fact that the T198P mutation significantly increases the neutralizing activity of the conformational antibodies, 1b12 and 48d. On the other hand, the IC9564 escape variant with the P198S mutation is resistant to these conformational antibodies and highly sensitive to the potent neutralizing antiserum, C1206, which recognizes a conformational epitope involving the sequences from V1, V2, and V3 regions in gp120. Thus, results from this study indicate that the conformation of gp120 can be exploited by HIV-1 to escape IC9564.