Acne vulgaris is a chronic inflammatory skin disorder commonly associated with the proliferation of pathogenic bacteria such as Cutibacterium acnes and Staphylococcus aureus. The increasing prevalence of antibiotic resistance has encouraged the exploration of natural resources as alternative antibacterial agents. This study aimed to formulate and evaluate an anti-acne cream containing esterified extract of stingless bee hive waste from Trigona laeviceps and to investigate its antibacterial activity against acne-causing bacteria. Stingless bee hive waste was extracted using ultrasound-assisted extraction with 96% ethanol, followed by sonochemical esterification using ethanol and propanol in the presence of sulfuric acid catalyst. The esterified extracts were characterized using FTIR and GC–MS analyses. Antibacterial activity of the esterified extracts was evaluated using the agar well diffusion method at concentrations of 10%, 15%, and 20% (w/v). The most active extract was subsequently formulated into an oil-in-water cream and evaluated for physicochemical properties, including pH, spreadability, adhesion, viscosity, homogeneity, and antibacterial activity. FTIR spectra confirmed successful esterification through the appearance of ester carbonyl absorption bands at approximately 1730–1740 cm⁻¹. GC–MS analysis revealed the presence of major bioactive compounds such as methyl oleate, ethyl oleate, squalene, lanosterol, cycloartenol, and 1-monooleoylglycerol. Ethanol-derived ester exhibited the strongest antibacterial activity, particularly at 20% concentration, with inhibition zones of 1.81 ± 0.10 cm against C. acnes and 1.94 ± 0.17 cm against S. aureus. The cream formulations showed good homogeneity, acceptable pH values, appropriate spreadability, and enhanced adhesion properties. The 20% cream formulation demonstrated the highest antibacterial activity against S. aureus with an inhibition zone of 1.64 ± 0.06 cm. In conclusion, esterified stingless bee hive waste, particularly ethanol-derived ester, possesses promising antibacterial potential and can be successfully formulated into a stable anti-acne cream for topical application.