The invention discloses preparation and application of a 5-aminolevulinic acid-camptothecin micromolecular prodrug. A preparation method comprises the following steps: (1) preparing a disulfide bond-containing camptothecin monomer BHD-CPT; (2) preparing CPT-SS-NPC; and (3) reacting a photosensitizer ALA with CPT-SS-NPC to prepare CPT-SS-ALA. Benefited from the amphiphilic structure of the drug conjugate, the drug conjugate can be self-assembled in water to form nanoparticles, and has the advantages of high micelle stability, controllable micelle shape, high drug loading capacity, low toxic andside effects, good drug controlled release and the like. In a reductive tumor microenvironment, disulfide bonds in the drug nanoparticles are broken, a chemotherapeutic drug CPT is released, meanwhile, the photosensitizer ALA continues to generate singlet oxygen under laser irradiation to kill cancer cells, and multi-modal synergistic treatment of tumors is achieved.