Phytotoxic substances in plants that may serve as alternative natural herbicides for controlling weeds are required for sustainable agriculture. We explored the phytotoxic activities of aqueous methanol extracts of Senna garrettiana (Craib) Irwin & Barneby leaves and the active substances they contain. The results revealed that the S. garrettiana leaf extracts had significant phytotoxic effects on three dicotyledons (Lepidium sativum L., Lactuca sativa L., and Medicago sativa L.) and two monocotyledons (Phleum pratense L. and Lolium multiflorum Lam.). An bioassay-guided isolation process yielded three active substances; caffeic acid, methyl caffeate, and (S)-6-hydroxymellein inhibited the seed germination, seedling growth, and biomass accumulation of L. sativum in a concentration-dependent manner. Based on the concentration required for 50% growth inhibition (IC50), (S)-6-hydroxymellein had the highest inhibitory effects on L. sativum in all test parameters, followed by methyl caffeate and caffeic acid. The L. sativum roots were the most susceptible to (S)-6-hydroxymellein (IC50 = 383 µM) and caffeic acid (IC50 = 2627 µM), whereas methyl caffeate (IC50 = 1361 µM) had the greatest effect on the L. sativum shoots. Thus, three isolated compounds may explain the phytotoxic effects of the S. garrettiana extracts. Consequently, caffeic acid, methyl caffeate, and (S)-6-hydroxymellein could be potential candidates for the future production of bioherbicides.