Introduction: To date, lung cancer has become increasingly prevalent and remains the leading cause of cancer-related death in both sexes, globally. Despite the advances of cancer treatment, systemic chemotherapy remains as the major treatment option for lung cancer. Nevertheless, the trend of chemotherapy resistance has restricted the efficacy of the chemotherapy treatment, thus leading the urge to develop an alternative chemotherapeutic agent which could give a promising treatment effect. Short peptides have acquired increasing interest as promising therapeutics due to its anticancer potential, rapid kinetics, high potency and low biocompatibility issue. In search of novel anti-cancer leads, the main objective of this study is to evaluate the in vitro antiproliferative properties of hybridized peptide analogues against human lung adenocarcinoma (A549) cell line. Methods: ND and DN analogues were designed based on two parent peptides, NDC1 and NDC2, through fragments hybridization approach. Modification of amino acid residues at specific positions of NDC1 and NDC2 was done at the C-terminal. Then, MTT assay was performed to examine the antiproliferative activities of NDC1, NDC2, NDs and DNs against A549 cells at concentrations ranging from 2-256μg/mL for 24 hours. Results: Findings obtained showed that the parental peptides, NDC1 and NDC2, exhibited IC50 values of 47.5±4.950μg/mL and 239±9.899μg/mL, respectively. All NDs showed excellent antiproliferative activities with IC50 values ranging from 22-71μg/mL. Nevertheless, all DNs did not display antiproliferative activity when tested up to 256μg/mL. We speculated that increased valine and isoleucine with decreased aspartic acid composition in NDs might be associated with their intermediate cytotoxicity strength, comparing with the parent peptides. However, the location of other amino acids in the peptide sequence should still be further investigated as it contributes to the peptide structure, hence leading to its selectivity and potency. Conclusion: As a conclusion, NDs could be further explored to develop a potent anti-cancer therapeutic drug.