Muto Hiroki, Kubara Kenji, Hitaoka Seiji, Osada Yoshihide, Suzuki Yuta
Tsukuba Research Laboratories, Eisai Co., Ltd., Ibaraki 300-2635, Japan.
Tsukuba Research Laboratories, Eisai Co., Ltd., Ibaraki 300-2635, Japan.
J Control Release. 2025 Jul 21;385:114055. doi: 10.1016/j.jconrel.2025.114055.
Lipid nanoparticles (LNPs) are clinically validated carriers for delivering drug agents for liver diseases. However, designing LNPs for delivery to hepatic stellate cells (HSCs) remains challenging. Here, using 12 ionizable lipids, including the approved MC3, SM-102, and ALC-0315, we demonstrate that ionizable lipid chemistry in LNPs determines the efficiency of delivery to HSCs. Using siRNAs targeting genes expressed exclusively in mouse HSCs (Reln) and hepatocytes (Ttr), we formulated an siRNA cocktail with LNPs. Systemic administration of 0.3 mg/kg body weight siRNA cocktails to mice caused varying Reln suppression (0-80 %) across the 12 lipids with consistent Ttr suppression (>90 %). Among the physicochemical properties of ionizable lipids, the delivery efficiency to HSCs could be influenced by the dipole moment of the ionizable lipids. The best-performing ALC-0315 LNP exhibited dose-dependent activity in HSCs (siRNA ED ∼ 0.03 mg/kg body weight) and durable silencing (up to 88 %) over two weeks, without toxicity, at 2 mg/kg. A comparative study in wild-type and ApoE-deficient mice revealed that ALC-0315 LNP delivery to HSCs was either independent of or slightly dependent on ApoE. In primary human activated HSCs, ALC-0315 LNPs induced robust cellular uptake and effective knockdown of heat shock protein 47, a therapeutic target for liver fibrosis, compared with MC3 LNPs. These findings will contribute to the development of advanced ionizable lipid-based LNPs for delivery to HSCs without targeting ligands.
脂质纳米颗粒（LNPs）是用于递送治疗肝病药物的经临床验证的载体。然而，设计用于递送至肝星状细胞（HSCs）的LNPs仍然具有挑战性。在这里，我们使用12种可电离脂质，包括已获批的MC3、SM - 102和ALC - 0315，证明了LNPs中的可电离脂质化学性质决定了递送至HSCs的效率。我们使用靶向仅在小鼠HSCs（Reln）和肝细胞（Ttr）中表达的基因的小干扰RNA（siRNAs），用LNPs配制了一种siRNA混合物。对小鼠全身给药0.3 mg/kg体重的siRNA混合物，在这12种脂质中导致了不同程度的Reln抑制（0 - 80%），而Ttr抑制则保持一致（>90%）。在可电离脂质的物理化学性质中，递送至HSCs的效率可能受可电离脂质偶极矩的影响。表现最佳的ALC - 0315 LNP在HSCs中表现出剂量依赖性活性（siRNA ED约为0.03 mg/kg体重），并且在2 mg/kg时，两周内具有持久的基因沉默效果（高达88%），且无毒性。在野生型和载脂蛋白E缺陷型小鼠中的一项比较研究表明，ALC - 0315 LNP递送至HSCs要么不依赖于载脂蛋白E，要么仅轻微依赖于载脂蛋白E。在原代人活化HSCs中，与MC3 LNP相比，ALC - 0315 LNP诱导了强大的细胞摄取并有效敲低了热休克蛋白47，热休克蛋白47是肝纤维化的一个治疗靶点。这些发现将有助于开发先进的基于可电离脂质的LNPs，用于在无靶向配体的情况下递送至HSCs。
