Chronic myeloid leukemia (CML) is a clonal myeloproliferative neoplasm characterized by the uncontrolled proliferation of myeloid cells [1]. The Philadelphia (pH+) chromosome arises from a reciprocal translocation event involving the Abelson murine leukemia virus (Abl) gene on chromosome 9 and the breakpoint cluster region (Bcr) on chromosome 22 t(9; 22) (q34; q11) [2], [3]. Bcr-Abl is a chimeric oncoprotein associated with the pathogenesis of Philadelphia chromosome-positive human leukemia, resulting in dysregulation of tyrosine kinase activity [4]. In recent years, the discovery of Imatinib (IMA) as the first designed BCR-ABL tyrosine kinase inhibitor (TKI), along with the introduction of third-generation TKIs such as Ponatinib, has significantly improved the survival prospects for patients diagnosed with CML [5], [6]. However, prolonged use of TKIs significantly impacts the efficacy of targeted therapy for leukemia due to both primary and acquired drug resistance, particularly during the transition from chronic to accelerated and acute stages [7], [8], [9].

Piperlongumine (PL) is a naturally occurring alkaloid first isolated from the root of Long Piper, a traditional Chinese herbal medicine, in 1967 (Fig. 1A) [10], [11]. and traditionally employed for the treatment of various diseases, including antiplatelet aggregation [12], anti-inflammation [13], cardiovascular protection [14], and anti-tumor. Encouragingly, recent studies have demonstrated that it exhibited the selective targeting ability towards various cancer cells, involving mechanisms such as reactive oxygen species (ROS) production, apoptosis, necrosis, and autophagy, thereby exerting anti-tumor effects [15], [16], [17]. However, the existing literature lacks sufficient structural diversity of PL derivatives and fails to demonstrate satisfactory anti-tumor activity, necessitating further exploration in terms of design, synthesis, and mechanism elucidation.

The α, β-unsaturated ketone skeleton of Xanthohumol, a natural prenylated chalcone exhibiting diverse biological activities, is illustrated in Fig. 1B [18]. Meanwhile, PL is characterized by the presence of two α, β-unsaturated ketone functionalities, which facilitate covalent interaction with intracellular thiols in proteins and glutathione [19]. The C2-C3 and C7-C8 double bonds described in the literature serve as essential pharmacophores acting as Michael acceptors, contributing to the cytotoxicity of PL towards cancer cells [20]. Moreover, the incorporation of urea structural fragments in molecular design and optimization can enhance biological activity, augment selectivity, modulate physicochemical properties, and facilitate metabolic stability while eliminating toxic pharmacophores, among other benefits [21], [22], [23]. Urea-containing compounds are increasingly employed in pharmaceutical chemistry and drug design, exemplified by the clinical anti-tumor drugs Sorafenib and Tivozanib (Fig. 1B) [24], [25].

In this study, a series of PL analogs were designed and synthesized by linking aryl-urea groups (Fig. 1A). The anti-tumor activity of all the synthesized target compounds was evaluated against PC-3, K562, and A549 cell lines. Furthermore, the preliminary mechanism of action for compound C5 was investigated, focusing on reactive oxygen species (ROS) generation and apoptosis induction. Transcriptomic analysis revealed that the differentially expressed genes (DEGs) primarily exhibited alterations in apoptosis and the PI3K/AKT signaling pathway. Molecular docking demonstrated that C5 displayed favorable binding affinity and scores towards Imatinib-Bcr-AblWT and Ponatinib-Bcr-AblT315I kinase proteins.