Abstract：
Limited treatment options and development of resistance to targeted therapy within few months pose significant challenges in the treatment of BRAF-mutated malignant melanoma. Moreover, extensive angiogenesis and vasculogenic mimicry promote the rapid progression of disease. The purpose of this study was to develop a protein kinase C inhibitor anchored BRD4 PROTAC (ARV) loaded PEGylated nanoliposomes (LARPC). Palmitoyl-dl-carnitine chloride (PC) was used as a protein kinase C inhibitor to provide a cationic surface charge to LARPC. The formulation was characterized for particle size, zeta potential, drug release and various cell culture assays using HUVEC and vemurafenib resistant melanoma cells. The particle size of LARPC was found to be 105.25 ± 2.76 nm with a zeta potential of +26.6 ± 6.25 mV. Inhibition of angiogenesis was demonstrated by ARV and LARPC using human umbilical vein endothelial cells (HUVEC)-based matrigel basement membrane model. Additionally, LARPC demonstrated very low IC50 with promising inhibition of vasculogenic mimicry channel formation, cell migration as well as colony formation in vemurafenib-resistant melanoma cell lines. Hence, the outcome of this combination therapy indicated the suitability of LARPC as a potential and novel approach for eradicating vemurafenib-resistant melanoma.
摘要：
有限的治疗选择和几个月内对靶向治疗的耐药性的发展对BRAF突变的恶性黑色素瘤的治疗提出了重大挑战。此外，广泛的血管生成和血管生成拟态促进疾病的快速进展。这项研究的目的是开发蛋白激酶C抑制剂锚定的BRD4PROTAC（ARV）负载的聚乙二醇化纳米脂质体（LARPC）。棕榈酰-dl-肉碱氯化物(PC)用作蛋白激酶C抑制剂以向LARPC提供阳离子表面电荷。该制剂的粒度进行了表征，zeta电位,使用HUVEC和vemurafenib抗性黑色素瘤细胞的药物释放和各种细胞培养测定。发现LARPC的粒径为105.25±2.76nm,ζ电位为+26.6±6.25mV。使用基于人脐静脉内皮细胞（HUVEC）的基质胶基底膜模型，通过ARV和LARPC证明了血管生成的抑制作用。此外,LARPC表现出非常低的IC50，有望抑制血管生成拟态通道的形成，威罗菲尼耐药黑色素瘤细胞系的细胞迁移和集落形成。因此，这种联合治疗的结果表明,LARPC适合作为根除维罗非尼耐药黑色素瘤的一种潜在的新方法.
