Exploring the antitumor effect of curcumin-piperlongumine hybrid molecule (CP) on EGFR-TKI-resistant non-small cell lung cancer using network pharmacological analysis and experimental verification

Abstract

EGFR-tyrosine kinase inhibitor (TKI) therapy is the most effective targeted therapy for non-small cell lung cancer (NSCLC). However, drug resistance remains a significant factor in the failure of lung cancer therapy. In the present study, we utilize network pharmacology, molecular docking, in vitro and in vivo experiments to explore the targets and biological mechanisms of CP, a novel curcumin-piperlongumine hybrid molecule, in EGFR-TKI-resistant NSCLC cells. The results reveal that CP exhibits enhanced biological activity compared to its parent compounds. CP can effectively inhibit cell proliferation by arresting cell cycle in the G2/M phase and inducing apoptosis. Mechanistically, CP-induced apoptosis is partially mediated by PI3K/AKT signaling pathway. These findings highlight the potential of CP as a promising therapeutic agent for EGFR-TKI-resistant lung cancer therapy.

1. Introduction

Lung cancer is the most commonly diagnosed cancer and remains the leading cause of cancer-related death throughout the world [1]. About 85% of lung cancer cases are non-small cell lung cancer (NSCLC) [2]. The epidermal growth factor receptor gene EGFR is one of the key driver genes in NSCLC. EGFR mutations often lead to abnormal activation of downstream pathways such as PI3K/AKT/mTOR and MAPK pathways, which promote the proliferation, differentiation and migration of tumor cells [3]. Approximately 20% of NSCLC patients harbor activating mutations in EGFR. The exon 19 deletions and the L858R point mutation comprise more than 90% of EGFR activating mutations [4]. According to current treatment guidelines, EGFR tyrosine kinase inhibitors (EGFR-TKIs) are the first-line treatment for advanced NSCLC patients with activating EGFR mutations. AZD9291 (osimertinib) is an irreversible third-generation epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI), and exhibits high selectivity against activating EGFR mutations, especially the T790M mutation [5]. However, the occurrence of acquired resistance is inevitable. Patients who initially benefit from osimertinib have been found to eventually develop resistance after 18.9 months [6]. Therefore, there is an urgent need to develop novel treatment strategies against EGFR-TKI resistance for advanced NSCLC.

Previous studies have revealed that curcumin and piperlongumine possess good antitumor activity in many types of cancers [7–9]. In addition, curcumin and piperlongumine have been reported to be potent tyrosine kinase inhibitors. It has been suggested that curcumin possesses good anti-EGFR activity, and curcumin overcomes Lenvatinib resistance through suppression of PI3K/AKT pathway in hepatocellular carcinoma [10]. Furthermore, curcumin analogs could dramatically inhibit the expression and phosphorylation of EGFR and reverse the EGFR-TKI resistance in gefitinib-resistant NSCLC [11,12]. Piperlongumine has also been revealed to suppress cell proliferation and angiogenesis of hepatocellular carcinoma via the EGF/EGFR axis [13]. Additionally, piperlongumine could overcome doxorubicin resistance in leukemia by suppressing the PI3K/AKT signaling pathway [14]. More importantly, piperlongumine could inhibit the phosphorylation of AKT, and reverse resistance to cisplatin in NSCLC [15]. In our previous study, we successfully synthesized a novel curcumin-piperlongumine hybrid molecule, CP [16]. Date revealed that CP exhibited enhanced structural stability and safety compared with its parent compounds in lung cancer cells. However, whether CP has better antitumor activity in EGFR-mutant NSCLC and overcome EGFR-TKI resistance is still unknown.

Network pharmacology approach has been widely applied in the study of various traditional Chinese medicine treatment [17–19]. It provides a novel network model of “multiple targets, multiple effects, and complex diseases” to investigate the potential mechanism of herbal compounds or natural products [20]. Moreover, network pharmacology affords a better understanding of the pathogenesis of diseases and new strategies for personalized treatment [21]. In the present study, we first utilized network pharmacology method to elucidate target genes and signaling pathways of CP and then further investigated the antitumor efficiency of CP on AZD9291-resistant NSCLC. This study may provide a potential therapeutic drug for EGFR-TKI-resistant lung cancer.