Despite the extensive development of aptamers in basic research, only a limited number have successfully progressed to clinical trials. This limitation is primarily attributed to the inherent instability of aptamers’ conformation, resulting in low affinity, poor serum stability, and inconsistent potency, posing a significant challenge to their stabilization. Herein, we established a feasible strategy to develop staple aptamers using the predicted binding conformations and titration cross-linking (TTC) method. Through this strategy, we successfully synthesized various stapled sclerostin aptamers with over 70% yield. Importantly, we demonstrated that stapled aptamers significantly enhanced their affinity (approximately 20-fold) and serum stability (negligible degradation within 32 h). Moreover, in an osteogenesis imperfecta mouse model (Col1a2^+/G610C mice), the stapled aptamer (named c-0127OA) exhibited a potent antagonistic effect on sclerostin, leading to enhanced anabolic bone anabolic potential. Collectively, our established stapling strategy is effective in stabilizing aptamers’ conformation, with c-0127OA emerging as a promising therapeutic candidate for osteogenesis imperfecta.

中文翻译：

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开发具有受限构象的吻合适配子，用于成骨不完美疗法


尽管适配体在基础研究中得到了广泛的发展，但只有少数适配体成功进入临床试验。这种限制主要归因于适配子构象的固有不稳定性，导致亲和力低、血清稳定性差和效力不一致，对其稳定性构成重大挑战。在此，我们建立了一种可行的策略，使用预测的结合构象和滴定交联 （TTC） 方法开发主要适配体。通过这种策略，我们成功合成了各种固定的硬化蛋白适配体，产率超过 70%。重要的是，我们证明钉合体显着增强了它们的亲和力（约 20 倍）和血清稳定性（32 小时内降解可忽略不计）。此外，在成骨不全症小鼠模型 （Col1a2^+/G610C 小鼠） 中，装订合体 （命名为 c-0127OA） 对硬化蛋白表现出有效的拮抗作用，导致合成代谢骨合成代谢潜力增强。总的来说，我们建立的吻合策略可有效稳定适配子的构象，其中 c-0127OA 成为成骨不全症的有前途的治疗候选药物。