Trilaurin-based solid lipid nanoparticles (TL-SLNs) containing paclitaxel (PTX) were prepared by hot-melt high-pressure homogenisation and subsequent freezing and thawing. PTX-containing TL-SLNs showed 160.0 ± 15.8 nm of mean particle size and -43.9 mV of zeta potential. Scanning electron microscopy also revealed the spherical shape and submicron-size of the TL-SLNs. Differential scanning calorimetry measurement presented melting transition peak of TL in SLNs indicating the solidified state of the core lipid. The prepared TL-SLNs were physically stable without significant particle size changes at 4 °C for 2 months. The amounts of uptake into the human ovarian cancer cells, SKOV-3, were similar between PTX delivered in Cremophor EL-based formulation and TL-SLNs. In vitro and in vivo antitumour activity of PTX in TL-SLNs was comparable to the commercial Cremophor EL-based formulation in SKOV-3. These results suggest that PTX-loaded TL-SLNs have promising potential as an alternative parenteral formulation for PTX.