DeMaria Andrew H, Lee Jeoung Soo, Webb Ken
Microenvironmental Engineering Laboratory, Department of Bioengineering, Clemson University, Clemson, SC 29634, USA.
Drug Design, Development, and Delivery (4D) Laboratory, Department of Bioengineering, Clemson University, Clemson, SC 29634, USA.
Gels. 2025 Jan 2;11(1):27. doi: 10.3390/gels11010027.
Hypoxia-inducible factor-1α (HIF-1α) initiates the cellular response to low oxygen levels, making it an attractive target for stimulating therapeutic angiogenesis. Several small molecules have been identified that stabilize HIF-1α and activate the angiogenic signaling pathway. However, achieving therapeutic doses of bioactive small molecules in target tissues remains challenging. In this paper, we report the synthesis and characterization of a new macromolecular prodrug composed of the pro-angiogenic small molecule N-oxalylglycine conjugated to hyaluronic acid (HA-NOG). NOG was conjugated to HA by esterification, and release was significantly increased in the presence of degradative enzymes, esterase and hyaluronidase, compared to physiological buffer, confirming that the release of NOG is primarily enzymatically driven. Normal human dermal fibroblasts (NHDFs) cultured with HA-NOG exhibited HIF-1α accumulation in the cell nucleus and dose-dependent increases in mRNA expression levels of three direct HIF transcriptional targets. Conditioned medium from these cells stimulated endothelial cell tubulogenesis. As an initial evaluation of safety and possible side effects, HA-NOG was found not to significantly affect NHDF metabolic activity, proliferation, or collagen deposition. These studies demonstrate that HA-NOG releases NOG in response to cellular enzymatic activity, activating the HIF signaling pathway and culminating in the secretion of soluble factors that activate endothelial cells without adversely affecting other cellular metabolic pathways.
缺氧诱导因子-1α（HIF-1α）启动细胞对低氧水平的反应，使其成为刺激治疗性血管生成的一个有吸引力的靶点。已经鉴定出几种能稳定HIF-1α并激活血管生成信号通路的小分子。然而，在靶组织中达到生物活性小分子的治疗剂量仍然具有挑战性。在本文中，我们报道了一种由促血管生成小分子N-草酰甘氨酸与透明质酸（HA-NOG）偶联而成的新型大分子前药的合成与表征。通过酯化反应将NOG与HA偶联，与生理缓冲液相比，在降解酶、酯酶和透明质酸酶存在的情况下，释放量显著增加，证实NOG的释放主要由酶驱动。用HA-NOG培养的正常人皮肤成纤维细胞（NHDFs）在细胞核中表现出HIF-1α积累，并且三个直接HIF转录靶点的mRNA表达水平呈剂量依赖性增加。这些细胞的条件培养基刺激内皮细胞形成管状结构。作为对安全性和可能副作用的初步评估，发现HA-NOG不会显著影响NHDF的代谢活性、增殖或胶原蛋白沉积。这些研究表明，HA-NOG响应细胞酶活性释放NOG，激活HIF信号通路，并最终导致激活内皮细胞的可溶性因子的分泌，而不会对其他细胞代谢途径产生不利影响。
