Leveraging engineered yeast small extracellular vesicles serve as multifunctional platforms for effectively loading methyl salicylate through the “esterase-responsive active loading” strategy

IF 4.4 2区医学 Q1 PHARMACOLOGY & PHARMACY

European Journal of Pharmaceutics and Biopharmaceutics Pub Date : 2025-03-18 DOI:10.1016/j.ejpb.2025.114696

Tianhao Li , Yun Zhou , Haoran Wang , Junfeng Wang , Rong Lu

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引用次数: 0

Abstract

Small extracellular vesicles (sEVs) are a promising vehicle for drug delivery because of their good biocompatibility and nontoxicity. The drug loading and encapsulation efficiencies of them are not satisfactory. This is especially the case when drugs are loaded by co-incubation. In this situation, as the difference in drug concentration between the inside and outside of the membrane of ordinary sEVs decreases, the drugs cannot diffuse efficiently into the inside of the vesicles. As a result, the drug loading efficiency is low.

In this study, engineered yeast-derived small extracellular vesicles derived from Pichia pastoris X33 (XPP-sEVs) engineered with carboxylesterase 1 (CES1) were constructed using the “esterase-responsive active loading” method, which is based on the concept of prodrug design and guided by the strategy of immobilized enzymes, to improve the loading efficiency of methyl salicylate (MS) to about twice as much. This was achieved by engineering the CES1-contained small extracellular vesicles to catalyze the esterase hydrolysis reaction of MS to establish a continuous MS transmembrane concentration gradient for efficient loading of the active drugs, including methyl salicylate and its hydrolyzed active product salicylic acid.

The results showed that the enzyme activity of the CES1-sEVs group finally reached 7.88 ± 0.43 U/mL, and the drug loading efficiency was about doubled. The results of drug release from the engineered extracellular vesicles showed that the release of the drug reached equilibrium around 100 min-2 h, during which there was no sudden release of the MS, and the final amount of the drug released could be increased by 12.34 % compared with the emulsion dosage form of the MS.

Overall, the CES1-sEVs prepared in this study significantly improved the drug-loading efficiency of MS without affecting the anti-inflammatory activity of MS. The MS-CES1-sEVs prepared in this study are non-toxic and have a bright application prospect in the treatment of skin inflammation.

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小细胞外囊泡（sEVs）具有良好的生物相容性和无毒性，是一种很有前景的给药载体。但它们的药物负载和封装效率并不令人满意。当药物通过共孵育的方式装载时，情况尤其如此。在这种情况下，由于普通 sEVs 膜内外的药物浓度差减小，药物无法有效扩散到囊泡内部。因此，药物装载效率很低。本研究采用 "酯酶响应式主动装载 "方法，在原药设计理念和固定化酶策略的指导下，构建了由 Pichia pastoris X33 衍生的工程化酵母细胞外小囊泡（XPP-sEVs），用羧基酯酶 1（CES1）工程化，将水杨酸甲酯（MS）的装载效率提高了约两倍。具体做法是利用含有 CES1 的细胞外小囊泡催化 MS 的酯酶水解反应，建立连续的 MS 跨膜浓度梯度，以高效装载包括水杨酸甲酯及其水解活性产物水杨酸在内的活性药物。结果表明，CES1-sEVs 组的酶活性最终达到了 7.88±0.43 U/mL，药物负载效率提高了约一倍。工程细胞外囊泡的药物释放结果表明，药物释放在 100 min-2 h 左右达到平衡，其间没有出现 MS 的突然释放，与 MS 的乳剂剂型相比，药物的最终释放量可增加 12.34 %。总之，本研究制备的 CES1-sEVs 在不影响 MS 抗炎活性的前提下显著提高了 MS 的药物负载效率。本研究制备的 MS-CES1-sEVs 无毒，在治疗皮肤炎症方面具有广阔的应用前景。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

European Journal of Pharmaceutics and Biopharmaceutics 医学-药学

CiteScore

8.80

自引率

4.10%

发文量

211

审稿时长

36 days

期刊介绍： The European Journal of Pharmaceutics and Biopharmaceutics provides a medium for the publication of novel, innovative and hypothesis-driven research from the areas of Pharmaceutics and Biopharmaceutics. Topics covered include for example: Design and development of drug delivery systems for pharmaceuticals and biopharmaceuticals (small molecules, proteins, nucleic acids) Aspects of manufacturing process design Biomedical aspects of drug product design Strategies and formulations for controlled drug transport across biological barriers Physicochemical aspects of drug product development Novel excipients for drug product design Drug delivery and controlled release systems for systemic and local applications Nanomaterials for therapeutic and diagnostic purposes Advanced therapy medicinal products Medical devices supporting a distinct pharmacological effect.