Ultrasound (US) exposure of soft tissues, such as the skin, has been shown to increase permeability, enhancing the passage of drug molecules via passive processes such as diffusion. However, US regimes have not been exploited to enhance active convective transport of drug molecules from a donor layer, such as a gel, into another medium. A layered tissue-mimicking material (TMM) was used as a model for a drug donor layer and underlying soft tissue to test penetration of agents in response to a range of US parameters. Influence of agent molecular mass (3-2000 kDa), US frequency (0.256/1.1 MHz) and US pressure (0-10 MPa) on transport was characterised. Agents of four different molecular sizes were embedded within the TMM with or without cavitation nuclei (CN) and US applied to achieve inertial cavitation. Post-insonation, samples were analysed to determine the concentration and penetration distance of agent transported. US exposure substantially enhanced transport. At both US frequencies, enhancement of transport was significantly higher (p < 0.05) above the cavitation threshold, and CN reduced the pressure at which cavitation, and therefore transport, was achieved. Acoustic cavitation activity and related phenomena was the predominant transport mechanism, and addition of CN significantly enhanced transport within a range of clinically applicable acoustic pressures. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci.