Dual-template molecular imprinting enables fully synthetic phosphatidylserine-targeted cancer therapy.

Phosphatidylserine (PS) exposed on tumor cell membranes is an attractive target for cancer therapy, but current PS-targeting strategies depend largely on biologically derived ligands that suffer from calcium dependence, limited stability, immunogenicity risk, and manufacturing complexity. Here, we report a fully synthetic therapeutic platform based on dual-template molecular imprinting that integrates PS-targeted tumor recognition with pH-responsive doxorubicin (Dox) delivery in a single nanocarrier. By co-imprinting the PS polar head group and Dox, the resulting molecularly imprinted polymer nanoparticles (dt-MIP) generate orthogonal binding sites for membrane targeting and drug loading, achieving high binding capacities for PS and Dox (Qmax ∼ 180 mg g-1 and ∼37 mg g-1, respectively), high selectivity, minimal drug leakage at physiological pH, and accelerated release in acidic environments. The dt-MIP recognized surface-exposed PS with specificity comparable to Annexin V and enabled PS-dependent Dox delivery across multiple cell lines, with delivery efficiency correlating with the degree of PS externalization. In triple-negative breast cancer cells, Dox-loaded dt-MIP markedly enhanced intracellular drug accumulation and reduced cell viability to ∼8% at 200 μg mL-1 dt-MIP, compared with ∼70% for free Dox at the same dose. In an MDA-MB-231 xenograft model, dt-MIP achieved tumor-selective accumulation and substantially suppressed tumor growth while maintaining a favorable safety profile. These results establish dual-template molecular imprinting as a general strategy for constructing fully synthetic, target-directed cancer therapeutics, positioning PS-imprinted nanocarriers as an alternative to biologically derived targeting systems.