<p>Numerous efforts have been devoted to altering the dynamic covalent linkers between the drug structural units in polyprodrugs from the viewpoint of molecular structure; however, the effect of their aggregation states has not yet been explored. Here, the effect of aggregation states on the <i>in vitro</i> drug release and cytotoxicity was investigated using a pH/glutathione (GSH) co-triggered degradable doxorubicin (DOX)-based polyprodrug (PDOX) as a model, which was synthesized by the facile polymerization of a pH/GSH dual-triggered dimeric prodrug (DDOX_ss) and 2,2-dimethoxypropane (DMP) by forming acid-labile ketal bond. Owing to the pH/GSH dual-triggered disulfide/<i>α</i>-amide and acid-labile ketal linkers between the DOX structural units, the resultant PDOX exhibited excellent pH/GSH co-triggered DOX release. With a similar diameter, the PDOX-NPs1 nanomedicines <i>via</i> fast precipitation showed faster DOX release than PDOX-NPs2 <i>via</i> slow self-assembly, regardless of their polymerization degree (DP). The effect of aggregation states is expected to be a secondary strategy for a more desired tumor intracellular microenvironment-responsive drug delivery for tumor chemotherapy, in addition to the molecular structures of polyprodrugs as drug self-delivery systems (DSDSs).</p>