C. Eugene Bennett Department of Chemistry, West Virginia University, 217 Clark Hall, Prospect Street, P.O. Box 6045, Morgantown, WV 26506, United States.
Toxicon. 2013 Nov;74:83-91. doi: 10.1016/j.toxicon.2013.07.028. Epub 2013 Aug 7.
The pharmaceutical world has greatly benefited from the well-characterized structure-function relationships of toxins with endogenous biomolecules, such as ion-channels, receptors, and signaling molecules. Thus, therapeutics derived from toxins have been aggressively pursued. However, the multifunctional role of various toxins may lead to undesirable off-target effects, hindering their use as therapeutic agents. In this paper, we suggest that previously unsuccessful toxins (due to off-target effects) may be revisited with mixtures by utilizing the pharmacodynamic response to the potential primary therapeutic as a starting point for finding new targets to ameliorate the unintended responses. In this proof of principle study, the pharmacodynamic response of HepG2 cells to a potential primary therapeutic (deguelin, a plant-derived chemopreventive agent) was monitored, and a possible secondary target (p38MAPK) was identified. As a single agent, deguelin decreased cellular viability at higher doses (>10 μM), but inhibited oxygen consumption over a wide dosing range (1.0-100 μM). Our results demonstrate that inhibition of oxygen consumption is related to an increase in p38MAPK phosphorylation, and may only be an undesired side effect of deguelin (i.e., one that does not contribute to the decrease in HepG2 viability). We further show that deguelin's negative effect on oxygen consumption can be diminished while maintaining efficacy when used as a therapeutic mixture with the judiciously selected secondary inhibitor (SB202190, p38MAPK inhibitor). These preliminary findings suggest that an endogenous response-directed mixtures approach, which uses a pharmacodynamic response to a primary therapeutic to determine a secondary target, allows previously unsuccessful toxins to be revisited as therapeutic mixtures.
制药领域极大地受益于毒素与内源性生物分子(如离子通道、受体和信号分子)之间特征明确的结构-功能关系。因此,从毒素中衍生的治疗药物一直受到积极的探索。然而,各种毒素的多功能作用可能导致不理想的脱靶效应,从而阻碍其作为治疗剂的应用。在本文中,我们提出可以通过利用潜在主要治疗药物的药效反应作为发现新靶点以改善非预期反应的起点,重新研究以前因脱靶效应而不成功的毒素(由于脱靶效应)。在这项原理验证研究中,我们监测了 HepG2 细胞对潜在主要治疗药物(角叉菜胶,一种植物来源的化学预防剂)的药效反应,并确定了一个可能的次要靶点(p38MAPK)。作为单一药物,角叉菜胶在较高剂量(>10 μM)下降低细胞活力,但在较宽的剂量范围内(1.0-100 μM)抑制耗氧量。我们的结果表明,耗氧量的抑制与 p38MAPK 磷酸化的增加有关,并且可能只是角叉菜胶的一种不良副作用(即,不有助于降低 HepG2 活力的副作用)。我们进一步表明,当用作治疗混合物与明智选择的二级抑制剂(SB202190,p38MAPK 抑制剂)时,角叉菜胶对氧消耗的负面影响可以减弱,同时保持疗效。这些初步发现表明,一种内源性反应导向的混合物方法,利用主要治疗药物的药效反应来确定次要靶点,可以重新研究以前不成功的毒素作为治疗混合物。