Four new compounds, including three secolignans (1-3) and one tetrahydrofuran lignan (4), were isolated from the petroleum ether and EtOAc fractions of Peperomia heyneana. These compounds were accompanied by eight known secolignans, one known tetrahydrofuran lignan, one known cyclohexenone, and one known amide. The structures were elucidated mainly by 1D and 2D NMR and MS experiments, and the relative configurations by NOE techniques. Five compounds were evaluated for their inhibitory activities against HIV-1 in infected C8166 cells.