Brenneman Douglas E, Kinney William A, McDonnell Mark E, Ippolito Michael J, Ward Sara Jane
Kannalife Sciences, Inc* Pennsylvania Biotechnology Center.
Temple University.
Res Sq. 2024 Feb 28:rs.3.rs-3982851. doi: 10.21203/rs.3.rs-3982851/v1.
KLS-13019 was reported previously to reverse paclitaxel-induced mechanical allodynia in a mouse model of chemotherapy-induced peripheral neuropathy (CIPN). Recent studies demonstrated that paclitaxel-induced increases in inflammatory markers (GPR55, NLRP3 and IL-1b) of dorsal root ganglion (DRG) cultures were shown to be reversed by KLS-13019 treatment. The mechanism of action for KLS-13019-mediated reversal of paclitaxel-induced neuroinflammation now has been explored using GPR55 siRNA. Pretreatment of DRG cultures with GPR55 siRNA produced a 21% decrease of immunoreactive (IR) area for GPR55 in cell bodies and a 59% decrease in neuritic IR area, as determined by high content imaging. Using a 24-hour reversal treatment paradigm, paclitaxel-induced increases in the inflammatory markers were reversed back to control levels after KLS-3019 treatment. Decreases in these inflammatory markers produced by KLS-13019 were significantly attenuated by GPR55 siRNA co-treatment, with mean IR area responses being attenuated by 56% in neurites and 53% in cell bodies. These data indicate that the percentage decreases in siRNA-mediated attenuation of KLS-13019-related efficacy on the inflammatory markers were similar to the percentage knockdown observed for neuritic GPR55 IR area. Similar studies conducted with cannabidiol (CBD), the parent compound of KLS-13019, produced low efficacy (25%) reversal of all inflammatory markers that were poorly attenuated (29%) by GPR55 siRNA. CBD was shown previously to be ineffective in reversing paclitaxel-induced mechanical allodynia. The present studies indicated significant differences between the anti-inflammatory properties of KLS-13019 and CBD which may play a role in their observed differences in the reversibility of mechanical allodynia in a mouse model of CIPN.
先前有报道称，KLS-13019可逆转化疗诱导的周围神经病变（CIPN）小鼠模型中紫杉醇诱导的机械性异常性疼痛。最近的研究表明，KLS-13019治疗可逆转紫杉醇诱导的背根神经节（DRG）培养物中炎症标志物（GPR55、NLRP3和IL-1β）的增加。现在已使用GPR55 siRNA探索了KLS-13019介导的逆转紫杉醇诱导的神经炎症的作用机制。用GPR55 siRNA预处理DRG培养物后，通过高内涵成像测定，细胞体中GPR55的免疫反应性（IR）面积减少了21%，神经突IR面积减少了59%。采用24小时逆转治疗模式，KLS-3019治疗后，紫杉醇诱导的炎症标志物增加恢复到对照水平。GPR55 siRNA共处理显著减弱了KLS-13019引起的这些炎症标志物的减少，神经突中的平均IR面积反应减弱了56%，细胞体中减弱了53%。这些数据表明，siRNA介导的KLS-13019相关疗效对炎症标志物的减弱百分比与神经突GPR55 IR面积的敲低百分比相似。用KLS-13019的母体化合物大麻二酚（CBD）进行的类似研究显示，所有炎症标志物的逆转效果较低（25%），且GPR55 siRNA对其减弱效果较差（29%）。先前已证明CBD在逆转紫杉醇诱导的机械性异常性疼痛方面无效。本研究表明，KLS-与CBD的抗炎特性存在显著差异，这可能在它们在CIPN小鼠模型中机械性异常性疼痛可逆性的观察差异中起作用。
