Araújo Héverton Mendes, Moura Gabriel Acácio de, Rocha Yasmim Mendes, Pinheiro Gomes Cristian Vicson, Melo de Oliveira Valentina Nascimento E, Oliveira Ronaldo Nascimento de, Figueiredo Nicolete Larissa Deadame de, Magalhães Emanuel Paula, de Menezes Ramon R P P B, Nicolete Roberto
Post-Graduate Program in Pharmaceutical Sciences (PPGCF), Federal University of Ceará (UFC), Fortaleza, CE, Brazil.
Fundação Oswaldo Cruz, Fiocruz, Fiocruz Ceará, Eusébio, CE, Brazil.
Biochem Biophys Rep. 2025 Feb 10;41:101950. doi: 10.1016/j.bbrep.2025.101950. eCollection 2025 Mar.
Melanoma is the most aggressive and lethal type of skin cancer, responsible for approximately 60,000 deaths annually. The main strategy for treating melanoma is surgery to completely remove the lesion and its margins. However, for more advanced cases with a high recurrence rate, the preferred approach is to combine chemotherapy with immunotherapy treatments. Tumor-associated macrophages (TAMs) are the most abundant leukocytes in solid tumors. Current immunotherapy approaches target TAMs by inhibiting pro-tumoral TAMs and activating anti-tumoral TAMs, repolarizing them to the M1 phenotype. The antitumor and immunomodulatory activities of molecules derived from 1,2,4-oxadiazole, as demonstrated in the literature, highlight the potential of this class as a source of promising candidates for therapeutic applications. Thus, the present study aims to evaluate the antitumor and immunomodulatory effects of the synthetic derivative 1,2,4-oxadiazole, N-cyclohexyl-3-(3-methylphenyl)-1,2,4-oxadiazole-5-amine (1,2,4-oxadiazole derivative 2), in melanoma cells and murine Bone Marrow-Derived Macrophages (BMDMs). Cytotoxicity in B16-F10 and BMDMs cells was assessed using the (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) MTT method. 1,2,4-oxadiazole derivative 2 exhibited antiproliferative effects on both cell lines, being 2.6 times more selective for B16-F10. Necrosis was identified as the active induced death pathway. BMDMs isolated and exposed to 1,2,4-oxadiazole derivative 2 polarize to the M1 phenotype and induce TNF-α at a concentration of 64.34 μM. Exposure to melanoma murine supernatants also promotes M1 polarization. Supernatants containing traces of 1,2,4-oxadiazole derivative 2 (Supernatants B, C, and D) increased the percentage of M1 cells compared to Supernatant A, as well as elevated levels of nitrite, TNF-α, and IL-12. 1,2,4-oxadiazole derivative 2 combined with Supernatant A and 1,2,4-oxadiazole derivative 2 combined with LPS also resulted in higher M1 polarization, suggesting a synergistic effect on M1 polarization and TNF-α production. Our findings underscore the significance of the 1,2,4-oxadiazole compound class and highlight the potential of 1,2,4-oxadiazole derivative 2 as an antitumoral and immunotherapeutic agent.
黑色素瘤是最具侵袭性和致命性的皮肤癌类型，每年导致约60000人死亡。治疗黑色素瘤的主要策略是通过手术完全切除病灶及其边缘。然而，对于复发率高的更晚期病例，首选方法是将化疗与免疫治疗相结合。肿瘤相关巨噬细胞（TAM）是实体瘤中最丰富的白细胞。目前的免疫治疗方法通过抑制促肿瘤TAM和激活抗肿瘤TAM，使其重新极化至M1表型来靶向TAM。文献表明，1,2,4-恶二唑衍生分子的抗肿瘤和免疫调节活性突出了该类化合物作为有前景的治疗应用候选物来源的潜力。因此，本研究旨在评估合成衍生物1,2,4-恶二唑，N-环己基-3-（3-甲基苯基）-1,2,4-恶二唑-5-胺（1,2,4-恶二唑衍生物2）对黑色素瘤细胞和小鼠骨髓来源巨噬细胞（BMDM）的抗肿瘤和免疫调节作用。使用（3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐）MTT法评估B16-F10细胞和BMDM细胞的细胞毒性。1,2,4-恶二唑衍生物2对两种细胞系均表现出抗增殖作用，对B16-F10的选择性高2.6倍。坏死被确定为诱导的活性死亡途径。分离并暴露于1,2,4-恶二唑衍生物2的BMDM极化至M1表型，并在浓度为64.34μM时诱导TNF-α。暴露于黑色素瘤小鼠上清液也促进M1极化。与上清液A相比，含有痕量1,2,4-恶二唑衍生物2的上清液（上清液B、C和D）增加了M1细胞的百分比，同时亚硝酸盐、TNF-α和IL-12水平升高。1,2,4-恶二唑衍生物2与上清液A组合以及1,2,4-恶二唑衍生物2与LPS组合也导致更高的M1极化，表明对M1极化和TNF-α产生具有协同作用。我们的研究结果强调了1,2,4-恶二唑化合物类别的重要性，并突出了1,2,4-恶二唑衍生物2作为抗肿瘤和免疫治疗剂的潜力。
