Department of Chemistry, Indian Institute of Technology Guwahati, Guwahati 781039, Assam, India.
Department of Pharmacology & Toxicology, National Institute of Pharmaceutical Education & Research, Guwahati 781101, Assam, India.
J Mater Chem B. 2024 May 1;12(17):4248-4261. doi: 10.1039/d3tb02552g.
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 (HS). 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 HO 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 HS was investigated in HeLa cells using the HS-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.
长期使用非常常见的处方非甾体抗炎药（NSAIDs）常常与不良副作用相关，包括由于环氧化酶非选择性抑制引起的胃肠道溃疡。我们描述了一种炎症刺激响应型的开环荧光诊断前药 DCF-HS 的开发，用于辅助药物递送。在活性氧物种（ROS）的激活下，前药释放双氯芬酸 DCF（活性药物）和近红外荧光团 DCI-NH2 以及羰基硫（COS）。酶碳酸酐酶（CA）对 COS 的第二次激活产生硫化氢（HS）。前药通过多步有机合成方便地合成。紫外-可见和荧光研究表明，DCF-HS 对 HO 等 ROS 在水相中的选择性反应性，以及在生理条件下用开环近红外荧光报告分子对药物 DCF 的所需开环。此外，使用反相高效液相色谱法证实了荧光团 DCI-NH2 和药物 DCF 的释放。接下来在细胞培养基中研究了前药激活的兼容性。前药 DCF-HS 在代表性癌细胞系（HeLa）和巨噬细胞系（RAW 264.7）中直至 100 μM 浓度下是无毒的，表明其生物相容性。使用 HS 选择性探针 WSP2 研究了细胞内 ROS 介导的前药激活及其近红外染料 DCI-NH2 和 HS 的释放在 HeLa 细胞中的作用。使用 Western blot 分析研究了前药 DCF-HS 中活性药物 DCF 的抗炎活性，并与母体药物 DCF 进行了比较，发现活性药物在剂量依赖性方式下显著抑制 COX-2。最后，在炎症诱导的 Wister 大鼠模型中验证了前药的抗炎潜力和开环荧光。
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