Background: It remains as a big challenge to provide therapeutic regime for the non-G12C KRAS-mutant non-small cell lung cancer (NSCLC) patients. The strategy of co-inhibition of MEK/RTKs pathways via trametinib and anlotinib showed preliminary activity in non-G12C KRAS-mutant NSCLC. Methods: The phase I clinical trial (NCT04967079) was divided into 2 parts including part A and part B. The primary endpoint of part A was to determine the recommended phase 2 dose (RP2D) and the primary endpoint of part B was to evaluate the objective response rate (ORR). The secondary endpoints were progression-free survival (PFS), disease control rate (DCR) and safety. Results: The part A containing 13 patients showed that the RP2D is trametinib (2 mg) plus anlotinib (8 mg), the ORR is 69.2%, the PFS is 207 days, DCR is 92% and the rate of adverse events (AEs) ≥ grade 3 is 23%. The part B containing 20 patients showed high efficacy of this combinational therapy (trametinib (2 mg) plus anlotinib (8 mg)), with the ORR at 65%, the PFS is 330 days, the DCR at 100%, and the rate of AEs ≥ grade 3 at 35%. An integrative analysis for the part A plus part B (33 patients) indicated that the ORR is 66.7%, the PFS is 300 days, the DCR is 97% and the rate of adverse events (AEs) ≥ grade 3 is 30%. Conclusions: This study provides a potential combinational therapeutic strategy for those non-G12C KRAS-mutant lung cancer patients via oral administration of trametinib and anlotinib. Clinical trial information: NCT04967079.