Despite the success for targeted delivery in the body, the efficient release without side effects caused by residual drug remains a challenge. For reducing residual drug, the pH-responsive carriers were prepared by self-assembly from aromatic macrocycles, which were non-toxic and biocompatible. The inner surroundings of aromatic macrocycles could be protonated positively by acid inducing the separation of neighboring macrocycles. Thus, Dox-loaded carriers successfully inhibited the proliferation of carcinoma cells (HepG2 and 4T1) rather than normal cells (HL7702). The effects were further proved in vivo without systemic cytotoxicity. Notably, the responsive environment for drug release depended on the concentration of carriers. Particularly, drug release was promoted by carrier separation. Carrier 2 exhibited preferable anticancer efficacy than carrier 1 due to the efficient release of Dox by full separation of the carrier. Collectively, we have developed a novel strategy serving as a selective and controlled drug release platform for cancer therapeutics.