Aims: Endostatin is a type of human vascular endostatin with more stable and higher activity. This study sought to evaluate pharmacokinetics (PK), safety and preliminary efficacy between treatment with endostatin 4 hours intravenous infusion and 72 hours or 168 hours intravenous pump infusion in patients with advanced non-small cell lung cancer (NSCLC). Methods: A total of 24 patients with advanced NSCLC were randomly assigned to group A and B. All patients received a standard 21 days treatment cycle of platinum-based doublet chemotherapy (paclitaxel or pemetrexed) and endostatin. Endostatin was administered at 7.5 mg/m2/day for 4 hours from day 1 to day 14 of cycle 1. Starting from day 1 of cycle 2, endostatin was administered at 105 mg/m2/cycle for 72 hours (group A) or 168 hours (group B), respectively. The total treatment duration was 4 cycles, with the option to extend up to 6 cycles. Results: The PK exposure per cycle of endostatin 4 hours infusion and 72 hours or 168 hours pump infusion were comparable between group A and group B. The majority of adverse events during pump infusion were consistent with those observed during infusion and no endostatin-related serious adverse events were reported. The overall objective response rate (ORR) was 37.5%. Conclusion: The PK exposure between intravenous infusion and intravenous pump infusion at the same total cycle dose is similar. Both 72 hours and 168 hours intravenous pump infusion endostatin were well tolerated, and no new unexpected safety signals were observed.