Objective To prepare prostate specific membrane antigen (PSMA)-targeting peptide-modified lipid-polymer nanoparticles co-loaded with homoharringtonine and triptolide (HHT/TPL-PSMA-Lips), optimize its preparation process, and evaluate its in vitro anti-prostate cancer activity. Methods The solubility and oil-water partition coefficient (lgP) of HHT and TPL were determined, and PSMA-targeting peptide-modified PEGylated phospholipids were synthesized. HHT/TPL-PSMA-Lips was prepared by the emulsification-solvent evaporation method, and the formulation was optimized using central composite design-response surface methodology (CCD-RSM). Indicators including particle size, ζ potential, and encapsulation efficiency were investigated. The in vitro release behavior of the formulation in different media was evaluated. MTT assay and flow cytometry were used to assess the in vitro inhibitory effect and apoptosis-inducing activity of the nanoparticles on prostate cancer cells, while cell uptake assay was performed to verify the tumor cell targeting efficiency of the nanoparticles. Results HHT and TPL showed low solubility in various media and strong lipophilicity. The molecular weight of the synthesized PSMA-targeting peptide-modified PEGylated phospholipids was 4 643.8. HHT/TPL-PSMA-Lips prepared with the optimal formulation exhibited uniform particle size, with an average particle size of (209.12 ± 3.24) nm, a ζ potential of (-34.4 ± 2.5) mV, and a comprehensive encapsulation efficiency of (81.20 ± 0.40)% [HHT (80.73 ± 0.86)%, TPL (82.00 ± 0.40)%]. In vitro release experiments demonstrated that HHT/TPL-PSMA-Lips could effectively improve the solubility of HHT and TPL, and exhibite a certain pH-responsive release property. Cell experiments showed that the HHT/TPL combined administration group displayed stronger inhibitory effect on prostate cancer cells and better apoptosis-inducing activity than the single drug groups. HHT/TPL-Lips enhanced the anti-tumor activity, and the targeted modified nanoparticles (HHT/TPL-PSMA-Lips) showed the strongest inhibitory effect on prostate cancer cells. Cell uptake assay indicated that modification with PSMA-targeting peptide could effectively enhance the cellular uptake of the nanoformulation in tumor cells. Conclusion The prepared HHT/TPL-PSMA-Lips not only solves the solubility problem of the drugs but also significantly enhances tumor targeting and anti-tumor efficacy, providing a new approach for the development of new traditional Chinese medicine formulations for the treatment of prostate cancer.

homoharringtonine  /  triptolide  /  prostate cancer  /  targeted nanoparticle  /  anti-tumor