Non-small cell lung cancer (NSCLC), pre-dominant subtype of lung cancer, is a global disorder affecting millions worldwide. One of the early treatments for NSCLC was use of a first-generation tyrosine kinase inhibitor, Erlotinib (Erlo). However, chronic exposure to Erlo led to development of acquired drug resistance (ADR) in NSCLC, limiting the clinical use of Erlo. A potential approach to overcome development of ADR is a multi-drug therapy. It has been previously reported that Erlo and Quinacrine (QA), an anti-malarial drug, can work synergistically to inhibit tumor progression in NSCLC. However, the combination failed at clinical stages, citing lack of efficacy. In this study, an effort has been made to improve the efficacy of Erlo-QA combination via development of nanoformulations, known to enhance therapeutic efficacy of potent chemotherapies. Synergy between Erlo and QA was measured via estimating the combination indices (CI). It was seen that established combination of nanoformulations (CI: 0.25) had better synergy than plain drug solutions (CI: 0.85) in combination. Following extensive in-vitro testing, data were simulated in biologically relevant 3D tumor models. Two tumor models were developed for extensive in-vitro testing, 3D-Spheroids grown in ultra-low attachment culture plates for efficacy evaluation and a 5D-spheroid model in 5D-sphericalplate with capability of growing 750 spheroids/well for protein expression analysis. Extensive studies on these models revealed that combination of Erlo and QA nanoformulations overall had a better effect in terms of synergy enhancement as compared to plain drug combination. Further, effect of combinatorial therapy on molecular markers was evaluated on 5D-Sphericalplate leading to similar effects on synergy enhancement. Results from present study suggests that combination of nanoformulations can improve the synergy between Erlo and QA while reducing the overall therapeutic dose.

非小细胞肺癌（NSCLC）是肺癌的主要亚型，是一种全球性疾病，影响着全球数百万人。NSCLC的早期治疗方法之一是使用第一代酪氨酸激酶抑制剂厄洛替尼（Erlo）。但是，长期接触Erlo导致NSCLC获得性耐药（ADR）的发展，限制了Erlo的临床应用。克服ADR发展的潜在方法是多药疗法。先前已有报道，抗疟疾药物Erlo和Quinacrine（QA）可以协同作用来抑制NSCLC中的肿瘤进展。但是，由于缺乏疗效，该组合在临床阶段失败了。在这项研究中，已努力通过开发纳米制剂来改善Erlo-QA组合的功效，已知该纳米制剂可增强强效化学疗法的治疗功效。Erlo和QA之间的协同作用是通过估算组合指数（CI）来衡量的。可以看出，已建立的纳米制剂组合（CI：0.25）比普通药物溶液（CI：0.85）具有更好的协同作用。继广泛在体外测试中，在生物学相关的3D肿瘤模型中模拟了数据。开发了两种肿瘤模型用于广泛的体外测试中，在超低附着力培养板中生长的3D球体用于功效评估，在5D球形板中的5D球体模型具有每孔生长750个球体的能力，用于蛋白质表达分析。对这些模型的大量研究表明，与普通药物组合相比，Erlo和QA纳米制剂的组合在协同增效方面总体上具有更好的效果。此外，在5D-Sphericalplate上评估了组合疗法对分子标记物的作用，从而对协同增效产生了相似的作用。目前研究的结果表明，纳米制剂的组合可以改善Erlo和QA之间的协同作用，同时降低总体治疗剂量。