Yoshioka Yukitake, Yamamoto Syunsuke, Kusamori Kosuke, Nakayama Miyu, Fujita Hisashi, Goto Akihiko, Iwasaki Shinji, Nagata Tetsuya, Itakura Shoko, Kusuhara Hiroyuki, Yokota Takanori, Hirabayashi Hideki, Nishikawa Makiya
Laboratory of Biopharmaceutics, Faculty of Pharmaceutical Sciences, Tokyo University of Science, Noda, Chiba, Japan.
Center of Excellence for Drug Metabolism, Pharmacokinetics and Modeling, Preclinical and Translational Sciences, Research, Takeda Pharmaceutical Company Limited, Kanagawa, Japan.
J Control Release. 2025 Apr 10;380:787-799. doi: 10.1016/j.jconrel.2025.02.025. Epub 2025 Feb 18.
Heteroduplex oligonucleotide (HDO) is a novel oligonucleotide therapeutic consisting of an antisense oligonucleotide (ASO) and its complementary RNA. A recent report showed that cholesterol-conjugated HDO (Chol-HDO) exhibited antisense activity in various tissues, including the brain; however, little information is available on the pharmacokinetic and plasma protein-binding properties of HDO and Chol-HDO. In the present study, we investigated the tissue distributions of an ASO, HDO, and Chol-HDO in mice and rats after intravenous injection. Tissue distribution was evaluated by measuring the concentration of ASO in tissue samples using liquid chromatography and tandem mass spectroscopy. ASO and HDO disappeared rapidly from the plasma, whereas Chol-HDO showed prolonged retention in the systemic circulation. The amount of ASO in the brain tissue was highest after injection of Chol-HDO, confirming its efficient delivery to the brain. The tissue distribution of oligonucleotides differed less in rats than in mice. Hepatic uptake of ASO and HDO, but not of Chol-HDO, was significantly inhibited by co-administration with the scavenger receptor inhibitor dextran sulfate sodium. The binding to plasma proteins was evaluated. Compared to ASO, HDO showed lower protein binding, but Chol-HDO showed much higher binding, with remarkable differences in binding to high-density and low-density lipoproteins. The binding of Chol-HDO to these proteins was also confirmed in mouse plasma after injection. These results indicate that the binding of Chol-HDO to plasma proteins, especially lipoproteins, is critical for determining tissue distribution and brain delivery after intravenous injection.
异源双链寡核苷酸（HDO）是一种新型寡核苷酸治疗药物，由反义寡核苷酸（ASO）及其互补RNA组成。最近的一份报告显示，胆固醇偶联的HDO（Chol-HDO）在包括脑在内的各种组织中均表现出反义活性；然而，关于HDO和Chol-HDO的药代动力学和血浆蛋白结合特性的信息却很少。在本研究中，我们研究了静脉注射后ASO、HDO和Chol-HDO在小鼠和大鼠体内的组织分布。通过使用液相色谱和串联质谱法测量组织样品中ASO的浓度来评估组织分布。ASO和HDO迅速从血浆中消失，而Chol-HDO在体循环中的保留时间延长。注射Chol-HDO后，脑组织中ASO的含量最高，证实其能有效递送至脑。寡核苷酸的组织分布在大鼠中比在小鼠中差异更小。与清除剂受体抑制剂硫酸葡聚糖钠共同给药可显著抑制肝脏对ASO和HDO的摄取，但对Chol-HDO无此作用。评估了与血浆蛋白的结合情况。与ASO相比，HDO显示出较低的蛋白结合，但Chol-HDO显示出更高的结合，在与高密度和低密度脂蛋白的结合方面存在显著差异。注射后在小鼠血浆中也证实了Chol-HDO与这些蛋白的结合。这些结果表明，Chol-HDO与血浆蛋白尤其是脂蛋白的结合对于静脉注射后确定组织分布和脑递送至关重要。
