Therapeutic nanoparticles (NPs) self-assembled from small molecular (pro)drug entities, opens up novel avenues for the generation of a wide range of drug delivery systems. Particularly, cocktail NPs created by co-assembly of multiple therapeutics often show profound efficacy beyond their individual agents. However, fabrication of synergistic NPs with high reproducibility and capability to deliver multiple therapeutics in a predefined ratio remains a challenge, which deters NP therapeutics from further clinical translation. In this work, a simple but versatile strategy has been developed to combine drug reconstitution and supramolecular nanoassembly to prodrug cocktail nanoparticle fabrication with microfluidics. Prodrugs reconstructed by PUFAylation were self-assembled into hybrid nanoparticles via microfluidic chip to synergistically deliver two chemotherapeutic drugs, 7-ethyl-10-hydroxy camptothecin (SN38) and paclitaxel (PTX), in a single nanoparticle container. In vitro cell-based assays demonstrate that the combinatorial chemotherapy is superior to each prodrug used alone while reduces the dosage of both drugs at the same time. Furthermore, the double-drug combination suppresses colon tumors by 86% at a total dosage of 16.7 mg/kg through synergy, and histological analysis indicates the safety of the hybrid nanoparticles. In general, this work shows that the nanomedicine synthesized by microfluidics provides considerable advantages including better size control and reproducibility, and great potential in effective combination therapy. It is expected to be applied to the fabrication of more chemical agent combination for other cancer types.

由小分子（前体）药物实体自组装的治疗性纳米颗粒 (NPs) 为生成各种药物递送系统开辟了新途径。特别是，由多种疗法共同组装产生的鸡尾酒 NPs 通常表现出超越其单个药物的深远功效。然而，制造具有高重现性和以预定比例提供多种疗法的协同 NP 仍然是一个挑战，这阻碍了 NP 疗法的进一步临床转化。在这项工作中，我们开发了一种简单但通用的策略，将药物重组和超分子纳米组装与微流体技术结合到前药鸡尾酒纳米颗粒制造中。通过PUFAylation重建前药自组装成纳米颗粒混合通过微流控芯片在单个纳米颗粒容器中协同递送两种化疗药物，7-乙基-10-羟基喜树碱（SN38）和紫杉醇（PTX）。体外基于细胞的测定表明，联合化疗优于单独使用的每种前药，同​​时降低了两种药物的剂量。此外，通过协同作用，双药组合以 16.7 mg/kg 的总剂量抑制结肠肿瘤 86%，组织学分析表明混合纳米颗粒的安全性。总的来说，这项工作表明，通过微流体合成的纳米药物具有相当大的优势，包括更好的尺寸控制和重现性，以及在有效联合治疗方面的巨大潜力。