Conjugates based on cell-penetrating peptides (CPPs) are scientifically relevant owing to their structural complexity; their ability to enter cells and deliver drugs, labels, antioxidants, bioactive compounds, or DNA fragments; and, consequently, their potential for application in research and biomedicine. In this study, carboxyamidated fluorescently labeled conjugates FAM-GG-TAT(47-57)-NH₂ and FAM-PEG₆-TAT(47-57)-NH₂ and photosensitizer-labeled conjugate Ch_k-PEG₆-TAT(47-57)-NH₂ [where TAT(47-57) is the CPP, 5(6)-carboxyfluorescein is the (FAM) fluorophore, chlorin k (Ch_k) is the photosensitizer, and the dipeptide glycyl–glycine (GG) or hexaethylene glycol (PEG₆) is the spacer] were originally designed, prepared, and fully characterized. Practically, all chemical reactions of the synthetic steps (peptide synthesis, spacer incorporation, and conjugation) were microwave-assisted at 60 °C using optimized protocols to give satisfying yields and high-quality products. Detailed analyses of the conjugates using spectrofluorimetry and singlet oxygen detection showed that they display photophysical properties typical of FAM or Ch_k. Anticandidal activity assays showed that not only this basic property of TAT(47-57) was preserved in the conjugates but also that the minimal inhibitory concentration was slightly reduced for cells incubated with PS-bearing conjugate Ch_k-PEG₆-TAT(47-57)-NH₂. Overall, these results indicated that the synthetic approach on-resin assisted by microwaves at 60 °C is simple, straightforward, selective, metal-free, sufficiently fast, cleaner, and more cost-effective than those previously used for preparing this type of macromolecule. Furthermore, such new data show that microwaves at 60 °C and/or conjugation did not harm the integrity of the conjugates’ constituents. Therefore, FAM-GG-TAT(47-57)-NH₂, FAM-PEG₆-TAT(47-57)-NH₂, and Ch_k-PEG₆-TAT(47-57)-NH₂ have high potential for practical applications in biochemistry, biophysics, and therapeutics.

中文翻译：

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细胞穿透肽 TAT(47-57) 的荧光和光敏结合物：设计、60 °C 下微波辅助合成和特性

基于细胞穿透肽（CPP）的缀合物由于其结构复杂性而具有科学意义；它们进入细胞并输送药物、标签、抗氧化剂、生物活性化合物或 DNA 片段的能力；因此，它们在研究和生物医学中的应用潜力。在本研究中，羧酰胺化荧光标记缀合物 FAM-GG-TAT(47-57)-NH ₂和 FAM-PEG ₆ -TAT(47-57)-NH ₂和光敏剂标记缀合物 Ch _k -PEG ₆ -TAT(47) -57)-NH ₂ [其中 TAT(47-57) 是 CPP，5(6)-羧基荧光素是 (FAM) 荧光团，二氢卟酚 k (Ch _k ) 是光敏剂，二肽甘氨酰-甘氨酸 (GG)或六乙二醇（PEG ₆）是间隔基]最初是设计、制备和充分表征的。实际上，合成步骤（肽合成、间隔基掺入和缀合）的所有化学反应均使用优化方案在 60 °C 下进行微波辅助，以获得令人满意的产量和高质量的产品。使用荧光分光光度法和单线态氧检测对缀合物进行详细分析表明，它们表现出 FAM 或 Ch _k典型的光物理特性。_{抗念珠菌活性测定表明，缀合物中不仅保留了 TAT(47-57) 的基本特性，而且与带有 PS 的缀合物 Ch k} -PEG ₆ -TAT(47-)一起孵育的细胞的最小抑制浓度略有降低。57)-NH ₂。总体而言，这些结果表明，60°C 微波辅助的树脂合成方法比以前用于制备此类高分子的方法简单、直接、选择性、不含金属、足够快、更清洁且更具成本效益。此外，这些新数据表明，60°C 的微波和/或缀合不会损害缀合物成分的完整性。因此，FAM-GG-TAT(47-57)-NH ₂、FAM-PEG ₆ -TAT(47-57)-NH ₂和 Ch _k -PEG ₆ -TAT(47-57)-NH ₂具有很高的潜力用于生物化学、生物物理学和治疗学的实际应用。