Conventional chemotherapy suffers from non-specificity, lack of aqueous solubility and multidrug resistance. Tumor-targeting nanotherapeutics exhibit unique advantages in the delivery of drugs specifically into tumors. Here, folic acid (FA), methionine (Met) and a near infrared (NIR) fluorescence probe (cypate, ICG derivative) were all bio-conjugated to succinyl-chitosan (SC) micelles. An anti-cancer drug (paclitaxel, PTX) was loaded into the hydrophobic cores of the formed FA-Met-SC-ICG derivative (FMSCI) micelles, which were under 200 nm in size. In comparison with micelles containing a single targeting moiety (FA or Met), FA and Met co-mediated micelles presented excellent biocompatibility, much higher affinity for cancer cells and excellent tumor-specific distribution in tumor-bearing mice. In vivo anti-tumor activity demonstrated that PTX-loaded FMSCI provided favourable therapeutic efficacy for tumors. In this research, novel nanotherapeutics based on FMSCI loaded with an anti-cancer drug provide a promising nanocomposite for combined tumor-targeting imaging and therapy.