Department of Polymer Science & Engineering, University of Massachusetts, Amherst, MA 01003, United States.
Department of Chemistry, Johannes Gutenberg University, Mainz, Germany.
J Control Release. 2024 Jan;365:950-956. doi: 10.1016/j.jconrel.2023.12.005. Epub 2023 Dec 19.
The application and design of protein transduction domains (PTDs) and protein transduction domain mimics (PTDMs) have revolutionized the field of biomacromolecule delivery. Our group has previously synthesized block copolymer PTDMs with well-defined hydrophobic and cationic blocks via ring-opening metathesis polymerization (ROMP). We have optimized the balance of hydrophobicity and cationic density to intracellularly deliver model proteins, active proteins, and antibodies. Despite the presence of serine, threonine, and tyrosine in naturally occurring PTDs, synthetic analogs have yet to be studied in PTDMs. In our present work, we introduce different alcohol groups to our PTDM structures as a new design parameter. A library of nine novel PTDMs were synthesized to incorporate alcohol groups of varying structures and evaluated based on their ability to intracellularly deliver fluorescently labeled antibodies. One PTDM in this novel library, named PTDM4, incorporates alcohol groups in both the hydrophobic and cationic blocks and was found to be the best performing PTDM with almost twice the median fluorescence intensity of the delivered antibody and half the cationic density compared to our positive control, a PTDM thoroughly studied by our group. PTDM4 was further studied by intracellularly delivering the active enzyme, TAT-Cre Recombinase. The activity of TAT-Cre Recombinase delivered by PTDM4 was comparable to that of the positive control, again with half the cationic density. This study is one of the first to examine the effects of alcohol groups on intracellular antibody and active enzyme delivery.
蛋白转导结构域（PTDs）和蛋白转导结构域模拟物（PTDMs）的应用和设计彻底改变了生物大分子递释领域。我们小组之前通过开环复分解聚合（ROMP）合成了具有明确疏水性和阳离子性嵌段的嵌段共聚物 PTDMs。我们优化了疏水性和阳离子密度的平衡，以实现模型蛋白、活性蛋白和抗体的细胞内递释。尽管天然 PTDs 中存在丝氨酸、苏氨酸和酪氨酸，但合成类似物尚未在 PTDMs 中进行研究。在我们目前的工作中，我们将不同的醇基团引入到 PTDM 结构中作为新的设计参数。合成了一个包含九个新型 PTDM 的文库，这些 PTDM 包含不同结构的醇基团，并根据其将荧光标记抗体细胞内递释的能力进行评估。在这个新型文库中，一个名为 PTDM4 的 PTDM 同时在疏水性和阳离子性嵌段中引入了醇基团，它被发现是表现最好的 PTDM，与我们的阳性对照相比，递释的抗体的中位数荧光强度增加了近两倍，而阳离子密度则降低了一半。与我们小组深入研究的阳性对照相比，PTDM4 进一步通过细胞内递释活性酶 TAT-Cre 重组酶进行了研究。PTDM4 递释的 TAT-Cre 重组酶的活性与阳性对照相当，而阳离子密度仅为其一半。这项研究是首次研究醇基团对细胞内抗体和活性酶递释的影响的研究之一。
