A novel amphiphilic peptide PA (APPA) containing two functional motifs is designed to coordinate the targeting to integrin alpha v beta 3 and neuropilin-1(NRP-1) on cell membrane. The cooperation of the two motifs fulfills the penetration of APPA-based nanosystem deep into tumor tissue and accumulates there. The APPA can be easily synthesized and self-assembled into spherical nanoparticles in the presence of doxorubicin (DOX) (PAD), or can be used for construction of fluorescent nanoprobe in the presence of DiR (PA-DiR). In vitro and in vivo studies demonstrate that the self-assembled PAD and PA-DiR nanosystems show a surprisingly high affinity for and penetration into alpha v beta 3 and NRP-1 positive tumors. The PA-DiR nanoprobe can be specifically recruited to 4T1 xenograft tumors. The signal of PA-DiR enriched in the tumor was more than 10 times higher than that of the negative control PC-DiR. The high affinity and specificity of the PA-DiR nanoprobe in in vivo fluorescence imaging, the profound therapeutic efficacy of PAD nanosystem in solid tumor models, and the low toxicity of APPA-based nanosystem demonstrate the great potential of this well-designed and simple nanosystem for application in the diagnosis and therapy of alpha v beta 3 and NRP-1 positive tumors, with minimal side effects.