生物正交前药疗法提供了一种有趣的双组分系统，其特点是增强的循环稳定性和按需受控激活。目前的策略通常通过单一机制的靶向机制将前药或其补充激活剂递送至肿瘤，这无法实现所需的抗肿瘤功效和安全性。两种不同且正交的机制的协调应该克服实体瘤的层次异质性，以同时提高生物正交前药疗法的两种成分的递送效率。我们在此通过整合两种正交的、不依赖于受体的肿瘤靶向策略开发了一种双机制靶向生物正交前药疗法。我们首先利用内源性白蛋白转运系统生成原位白蛋白结合的生物正交笼阿霉素前药，具有延长的血浆循环和在肿瘤部位的选择性积累。然后，我们采用酶指导自组装（EISA）来特异性富集肿瘤细胞内的生物正交激活剂。由于每种靶向递送模式都会诱导内在的药代动力学特征，因此根据其药代动力学进一步优化给药顺序，可以实现时空控制的前药靶向和按需激活。总之，通过协调两种离散且不依赖于受体的靶向策略，我们开发了一种基于全小分子的生物正交前药系统，用于双机制靶向抗癌治疗，以最大限度地提高化疗药物的治疗效果并最大限度地减少药物不良反应。

Bioorthogonal prodrug therapies offer an intriguing two-component system that features enhanced circulating stability and controlled activation on demand. Current strategies often deliver either the prodrug or its complementary activator to the tumor with a monomechanism targeted mechanism, which cannot achieve the desired antitumor efficacy and safety profile. The orchestration of two distinct and orthogonal mechanisms should overcome the hierarchical heterogeneity of solid tumors to improve the delivery efficiency of both components simultaneously for bio-orthogonal prodrug therapies. We herein developed a dual-mechanism targeted bioorthogonal prodrug therapy by integrating two orthogonal, receptor-independent tumor-targeting strategies. We first employed the endogenous albumin transport system to generate the in situ albumin-bound, bioorthogonal-caged doxorubicin prodrug with extended plasma circulation and selective accumulation at the tumor site. We then employed enzyme-instructed self-assembly (EISA) to specifically enrich the bioorthogonal activators within tumor cells. As each targeted delivery mode induced an intrinsic pharmacokinetic profile, further optimization of the administration sequence according to their pharmacokinetics allowed the spatiotemporally controlled prodrug activation on-target and on-demand. Taken together, by orchestrating two discrete and receptor-independent targeting strategies, we developed an all-small-molecule based bioorthogonal prodrug system for dual-mechanism targeted anticancer therapies to maximize therapeutic efficacy and minimize adverse drug reactions for chemotherapeutic agents.