FGF21 is an endocrine signaling protein belonging to the family of fibroblast growth factors (FGFs). It has emerged as a molecule of interest for treating various metabolic diseases due to its role in regulating glucogenesis and ketogenesis in the liver. However, FGF21 is prone to heat, proteolytic, and acid-mediated degradation, and its low molecular weight makes it susceptible to kidney clearance, significantly reducing its therapeutic potential. Protein engineering studies addressing these challenges have generally shown that increasing the thermostability of FGF21 led to improved pharmacokinetics. Here, we describe the computer-aided design and experimental characterization of FGF21 variants with enhanced melting temperature up to 15°C, uncompromised efficacy at activation of MAPK/ERK signaling in Hep G2 cell culture, and ability to stimulate proliferation of Hep G2 and NIH 3T3 fibroblasts cells comparable with FGF21-WT. We propose that stabilizing the FGF21 molecule by rational design should be combined with other reported stabilization strategies to maximize the pharmaceutical potential of FGF21.

中文翻译：

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稳定成纤维细胞生长因子 21 的计算机辅助工程

FGF21 是一种内分泌信号蛋白，属于成纤维细胞生长因子 (FGF) 家族。由于其在调节肝脏糖生成和酮生成中的作用，它已成为治疗各种代谢疾病的重要分子。然而，FGF21 容易发生热、蛋白水解和酸介导的降解，而且其低分子量使其容易被肾脏清除，从而显着降低其治疗潜力。针对这些挑战的蛋白质工程研究普遍表明，提高 FGF21 的热稳定性可以改善药代动力学。在这里，我们描述了 FGF21 变体的计算机辅助设计和实验表征，其熔解温度提高至 15°C，在 Hep G2 细胞培养物中激活 MAPK/ERK 信号传导具有毫不妥协的功效，并且能够刺激 Hep G2 和 NIH 增殖3T3成纤维细胞与FGF21-WT相当。我们建议通过合理设计稳定 FGF21 分子应与其他报道的稳定策略相结合，以最大限度地发挥 FGF21 的药物潜力。