Settelmeier Stephan, Varasteh Zohreh, Staniszewska Magdalena, Beerlage Anna-Lena, Zarrad Fadi, Fendler Wolfgang P, Rischpler Christoph, Notni Johannes, Totzeck Matthias, Herrmann Ken, Rassaf Tienush, Hendgen-Cotta Ulrike B
Department of Cardiology and Vascular Medicine, West German Heart and Vascular Center, Medical Faculty, University Hospital Essen, University of Duisburg-Essen, 45147 Essen, Germany.
Department of Nuclear Medicine, Medical Faculty, University Hospital Essen, University of Duisburg-Essen, 45147 Essen, Germany.
Pharmaceuticals (Basel). 2023 May 31;16(6):824. doi: 10.3390/ph16060824.
The cardiac bioavailability of peptide drugs that inhibit harmful intracellular protein-protein interactions in cardiovascular diseases remains a challenging task in drug development. This study investigates whether a non-specific cell-targeted peptide drug is available in a timely manner at its intended biological destination, the heart, using a combined stepwise nuclear molecular imaging approach. An octapeptide (heart8P) was covalently coupled with the trans-activator of transcription (TAT) protein transduction domain residues 48-59 of human immunodeficiency virus-1 (TAT-heart8P) for efficient internalization into mammalian cells. The pharmacokinetics of TAT-heart8P were evaluated in dogs and rats. The cellular internalization of TAT-heart8P-Cy(5.5) was examined on cardiomyocytes. The real-time cardiac delivery of Ga-NODAGA-TAT-heart8P was tested in mice under physiological and pathological conditions. Pharmacokinetic studies of TAT-heart8P in dogs and rats revealed a fast blood clearance, high tissue distribution, and high extraction by the liver. TAT-heart-8P-Cy(5.5) was rapidly internalized in mouse and human cardiomyocytes. Correspondingly, organ uptake of hydrophilic Ga-NODAGA-TAT-heart8P occurred rapidly after injection with an initial cardiac bioavailability already 10 min post-injection. The saturable cardiac uptake was revailed by the pre-injection of the unlabeled compound. The cardiac uptake of Ga-NODAGA-TAT-heart8P did not change in a model of cell membrane toxicity. This study provides a sequential stepwise workflow to evaluate the cardiac delivery of a hydrophilic, non-specific cell-targeting peptide. Ga-NODAGA-TAT-heart8P showed rapid accumulation in the target tissue early after injection. The implementation of PET/CT radionuclide-based imaging methodology as a means to assess effective and temporal cardiac uptake represents a useful and critical application in drug development and pharmacological research and can be extended to the evaluation of comparable drug candidates.
在心血管疾病中，抑制有害细胞内蛋白质 - 蛋白质相互作用的肽类药物的心脏生物利用度在药物开发中仍然是一项具有挑战性的任务。本研究使用联合逐步核分子成像方法，调查一种非特异性细胞靶向肽类药物是否能及时到达其预期的生物学目的地——心脏。一种八肽（heart8P）与人免疫缺陷病毒 -1（HIV-1）转录激活因子（TAT）蛋白转导结构域的48 - 59位残基共价偶联（TAT-heart8P），以便有效地内化进入哺乳动物细胞。在犬和大鼠中评估了TAT-heart8P的药代动力学。在心肌细胞上检测了TAT-heart8P-Cy(5.5)的细胞内化情况。在生理和病理条件下，对小鼠进行了Ga-NODAGA-TAT-heart8P的实时心脏递送测试。TAT-heart8P在犬和大鼠中的药代动力学研究表明，其血液清除快、组织分布高且肝脏摄取高。TAT-heart-8P-Cy(5.5)在小鼠和人心肌细胞中迅速内化。相应地，注射亲水性Ga-NODAGA-TAT-heart8P后，器官摄取迅速发生，注射后10分钟时初始心脏生物利用度就已出现。预先注射未标记的化合物可使饱和的心脏摄取占优势。在细胞膜毒性模型中，Ga-NODAGA-TAT-heart8P的心脏摄取没有变化。本研究提供了一个顺序逐步的工作流程，以评估亲水性、非特异性细胞靶向肽的心脏递送。Ga-NODAGA-TAT-heart8P在注射后早期在靶组织中迅速积累。基于PET/CT放射性核素的成像方法作为评估有效和瞬时心脏摄取的手段，在药物开发和药理学研究中是一种有用且关键的应用，并且可以扩展到对类似候选药物的评估。
