Prolonged use of very commonly prescribed non-steroidal anti-inflammatory drugs (NSAIDs) is often associated with undesired side effects, including gastrointestinal ulcers due to the non-selective inhibition of cyclooxygenases. We describe the development of an inflammatory-stimuli-responsive turn-on fluorogenic theranostic prodrug DCF-HS for adjuvant drug delivery. Upon activation by reactive oxygen species (ROS), the prodrug releases diclofenac DCF (active drug) and the NIR fluorophore DCI-NH₂ along with carbonyl sulfide (COS). The second activation of COS by the enzyme carbonic anhydrase (CA) generates hydrogen sulfide (H₂S). The prodrug was conveniently synthesized using multi-step organic synthesis. The UV-Vis and fluorescence studies revealed the selective reactivity of DCF-HS towards ROS such as H₂O₂ in the aqueous phase and the desired uncaging of the drug DCF with turn-on NIR fluorescent reporter under physiological conditions. Furthermore, the release of fluorophore DCI-NH₂ and drug DCF was confirmed using the reverse phase HPLC method. Compatibility of prodrug activation was studied next in the cellular medium. The prodrug DCF-HS was non-toxic in a representative cancer cell line (HeLa) and a macrophage cell line (RAW 264.7) up to 100 μM concentration, indicating its biocompatibility. The intracellular ROS-mediated activation of the prodrug with the release of NIR dye DCI-NH₂ and H₂S was investigated in HeLa cells using the H₂S-selective probe WSP2. The anti-inflammatory activity of the active drug DCF from the prodrug DCF-HS was studied in the lipopolysaccharide (LPS)-induced macrophage cell line and compared to that of the parent drug DCF using western blot analysis and it was found that the active drug resulted in pronounced inhibition of COX-2 in a dose-dependent manner. Finally, the anti-inflammatory potential of the prodrug and the turn-on fluorescence were validated in the inflammation-induced Wister rat models.

中文翻译：

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炎症刺激响应开启近红外荧光治疗诊断前药：双氯芬酸和硫化氢的辅助递送可减轻急性炎症性疾病

长期使用非常常用的非甾体类抗炎药 (NSAID) 通常会产生不良副作用，包括由于环氧合酶的非选择性抑制而导致的胃肠道溃疡。我们描述了用于辅助药物递送的炎症刺激响应开启荧光治疗诊断前药DCF-HS的开发。被活性氧 (ROS) 激活后，前药释放双氯芬酸DCF（活性药物）和近红外荧光团DCI-NH ₂以及羰基硫 (COS)。碳酸酐酶 (CA) 对 COS 的第二次激活产生硫化氢 (H ₂ S)。使用多步有机合成方便地合成前药。紫外-可见光和荧光研究揭示了DCF-HS对水相中的ROS（例如 H ₂ O _{2 ）}的选择性反应性，以及在生理条件下通过打开近红外荧光报告基因对药物DCF进行所需的解封。此外，使用反相HPLC方法确认了荧光团DCI-NH ₂和药物DCF的释放。接下来在细胞介质中研究前药激活的相容性。前药DCF-HS在浓度高达 100 μM 的代表性癌细胞系 (HeLa) 和巨噬细胞系 (RAW 264.7) 中是无毒的，表明其具有生物相容性。使用 H ₂ S 选择性探针WSP2在 HeLa 细胞中研究了细胞内 ROS 介导的前药激活以及近红外染料DCI-NH ₂和 H ₂ S的释放。在脂多糖（LPS）诱导的巨噬细胞系中研究了前药DCF-HS的活性药物DCF的抗炎活性，并使用蛋白质印迹分析与母体药物DCF进行比较，发现活性药物以剂量依赖性方式导致 COX-2 的显着抑制。最后，前药的抗炎潜力和开启荧光在炎症诱导的 Wister 大鼠模型中得到了验证。