Lung cancer claims the highest mortality and the second-most estimated new cases among all oncological diseases [1]. Non-small cell lung cancer (NSCLC) accounts for approximately 85% of all newly diagnosed lung cancers [2]. Approximately 40% of newly diagnosed lung cancer patients are stage IV. For stage IIIB/IV NSCLC, cytotoxic combination chemotherapy is standard first-line chemotherapy. A regimen of platinum (cisplatin or carboplatin) plus paclitaxel, gemcitabine, docetaxel, vinorelbine, irinotecan, or pemetrexed is the recommended clinical treatment [3]. Cisplatin is the first-generation platinum-based anti-cancer drug. Although cisplatin is much more effective than other platinum drugs at the same dosage [4], accumulating reports have shown the failure of conventional platinum-based chemotherapy due to various side effects and drug resistance [5]. Miriplatin, a member of platinum drug family, has been approved in Japan in 2009 for transcatheter arterial chemoembolization treatment of hepatocellular carcinoma (HCC) [6]. Miriplatin is a lipophilic platinum drug that contains myristates (14-carbon chains) as leaving groups and diamino cyclohexane as the non-leaving carrier ligand. The application of miriplatin in clinic is limited because it has very poor solubilities both in water and in common organic solvents [7]. The structure of solid tumors and tumor microenvironment (TME) in lung cancer constitute a barrier to the deep penetration of chemotherapy agents, which limits the effectiveness of chemotherapy [8]. Nanoparticles with appropriate properties provide a promising delivery system to overcome the biological and physiochemical barriers that hinder anti-cancer activity [9]. Lipid-based nanoparticles such as liposomes, micelles, and solid lipid nanoparticles (SLNs) can delivery anti-cancer drugs to improve their anti-cancer activities. In this study, we formulated miriplatin into various micelles, liposomes, and SLNs by film-hydration and evaluated their physicochemical properties and anti-cancer activity against NSCLC cells in culture. Miriplatin-loaded formulations with different compositions were successfully prepared by the film-hydration method. Most miriplatin-loaded micelles were more homogeneous and much smaller than miriplatin-loaded liposomes and SLNs. The majority of miriplatin-loaded micelles were about 15 nm in diameter, while SLNs were around 120 nm, and liposomes were about 180 nm. Formulations with a higher molar ratio of PE-PEG2000 had smaller sizes. SLNs loaded with a higher molar ratio of miriplatin in the compositions showed smaller sizes. Inductively coupled plasma mass spectrometry (ICP-MS) and inductively coupled plasma optical emission spectrometry (ICP-OES) techniques were attempted to quantify the platinum element in the formulations. Formulations with a higher molar ratio of PE-PEG2000 had higher recovery of platinum element. Most miriplatin-loaded formulations had higher than 80% platinum recovery. The recovery of intact miriplatin was characterized by HPLC. Miriplatin-loaded micelles had much higher intact miriplatin recovery (about 100%) than SLNs (about 30%). By TEM imaging, the micelles showed the morphology of spherical dots of about 10 nm in diameter while SLNs showed both spherical and rodlike structures of about 120 nm in diameter. The TEM results were consistent with the size and PDI results by the Zetasizer. Three-dimensional multicellular spheroids (3D MCS) of A549 and A549-iRFP cell lines were successfully established as cell culture models to evaluate activity against non-small cell lung cancer. The viability of 3D MCS after 7-days treatment with miriplatin-loaded micelles was about 0%, which was similar to cisplatin. Miriplatin-loaded formulations with a higher molar ratio of PE-PEG2000 in the compositions had higher anti-cancer activity against 3D MCS. The anticancer activity of miriplatin-loaded formulations against 3D MCS was positively associated with the recovery of intact miriplatin from the formulations. The IC50 value of miriplatin-loaded micelles against A549-iRFP 3D MCS was around 25 µM, while that of cisplatin was 84.78 µM. In summary, the reported lipid-based nano-formulations represent a promising delivery system of miriplatin against NSCLC.