Dilinoleylmethyl-4-dimethylaminobutyrate (D-Lin-MC3-DMA) are potent siRNA delivery vehicles in vivo[1].
D-Lin-MC3-DMA is an effective carrier for delivering siRNA in HeLa cells[1]. Differences in gene silencing in two distinct liver cell populations when siRNA was delivered with LNPs containing the ionizable cationic lipid MC3(D-Lin-MC3-DMA) or ALC-0315[1]. D-Lin-MC3-DMA, which is practically uncharged at physiological pH, can become ionized once endocytosed and entrained in the early endosome compartment. Because these early endosomes are expected to contain negatively charged phosphatidylserine lipids, one may speculate that the LNP cargo might escape by LNPs adhering to and fusing with the early endosomal membrane through electrostatic means[5,6]. D-Lin-MC3-DMA -LNPs were close to neutral charge with a zeta-potential of 5±3 mV. While D-Lin-MC3-DMA -LNPs are neutrally charged at physiological pH, they are known to be positively charged at endosomal pH[7]. LNPs were synthesized by the microfluidic mixing technique and are composed of ionizable cationic lipid (D-Lin-MC3-DMA), a phospholipid, cholesterol, and poly(ethylene glycol) (PEG), as well as encapsulated cargoes that are either phosphorothioated siRNA (50 or 100%) or mRNA. LNPs form physically stable complexes with bioactive drug siRNA for a period of 94 days[8].
In the murine FVII model, an ED50 of 0.005 mg kg 1 siRNA was achieved in the optimized composition(D-Lin-MC3-DMA), which represented a sixfold improvement relative to the 40/10/40/10 molar ratio composition[2]. 2nd generation LNP containing D-Lin-MC3-DMA were more than two orders of magnitude more potent than 1st generation LNP containing DLinDMA[4].
References:
[1]. Kulkarni JA, Myhre JL, et,al.Design of lipid nanoparticles for in vitro and in vivo delivery of plasmid DNA. Nanomedicine. 2017 May;13(4):1377-1387. doi: 10.1016/j.nano.2016.12.014. Epub 2016 Dec 28. PMID: 28038954.
[2]. Jayaraman M, Ansell SM, et,al. Maximizing the potency of siRNA lipid nanoparticles for hepatic gene silencing in vivo. Angew Chem Int Ed Engl. 2012 Aug 20;51(34):8529-33. doi: 10.1002/anie.201203263. Epub 2012 Jul 10. PMID: 22782619; PMCID: PMC3470698.
[3]. Ferraresso F, Strilchuk AW, et,al.Comparison of DLin-MC3-DMA and ALC-0315 for siRNA Delivery to Hepatocytes and Hepatic Stellate Cells. Mol Pharm. 2022 Jul 4;19(7):2175-2182. doi: 10.1021/acs.molpharmaceut.2c00033. Epub 2022 May 31. PMID: 35642083; PMCID: PMC9621687.
[4]. Akinc A, Maier MA, et,al. The Onpattro story and the clinical translation of nanomedicines containing nucleic acid-based drugs. Nat Nanotechnol. 2019 Dec;14(12):1084-1087. doi: 10.1038/s41565-019-0591-y. PMID: 31802031.
[5]. Yanez Arteta M, Kjellman T, et,al. Successful reprogramming of cellular protein production through mRNA delivered by functionalized lipid nanoparticles. Proc Natl Acad Sci U S A. 2018 Apr 10;115(15):E3351-E3360. doi: 10.1073/pnas.1720542115. Epub 2018 Mar 27. PMID: 29588418; PMCID: PMC5899464.
[6]. Chan CL, Majzoub RN, et,al. Endosomal escape and transfection efficiency of PEGylated cationic liposome-DNA complexes prepared with an acid-labile PEG-lipid. Biomaterials. 2012 Jun;33(19):4928-35. doi: 10.1016/j.biomaterials.2012.03.038. Epub 2012 Apr 1. PMID: 22469293; PMCID: PMC3337860.
[7]. Maugeri M, Nawaz M, et,al. Linkage between endosomal escape of LNP-mRNA and loading into EVs for transport to other cells. Nat Commun. 2019 Sep 24;10(1):4333. doi: 10.1038/s41467-019-12275-6. PMID: 31551417; PMCID: PMC6760118.
[8]. Viger-Gravel J, Schantz A, et,al.Structure of Lipid Nanoparticles Containing siRNA or mRNA by Dynamic Nuclear Polarization-Enhanced NMR Spectroscopy. J Phys Chem B. 2018 Feb 22;122(7):2073-2081. doi: 10.1021/acs.jpcb.7b10795. Epub 2018 Feb 9. PMID: 29332384.
Dilinoleylmethyl-4-dimethylaminobutyrate (D-Lin-MC3-DMA) 是有效的体内 siRNA 递送载体[1]。
D-Lin-MC3-DMA 是在 HeLa 细胞中递送 siRNA 的有效载体[1]。当 siRNA 与含有可电离阳离子脂质 MC3(D-Lin-MC3-DMA) 或 ALC-0315[1] 的 LNP 一起递送时,两个不同肝细胞群中基因沉默的差异。 D-Lin-MC3-DMA 在生理 pH 下几乎不带电荷,一旦被内吞并夹带在早期内体隔室中,就会电离。由于预计这些早期内体含有带负电荷的磷脂酰丝氨酸脂质,因此可以推测 LNP 货物可能通过静电方式粘附并与早期内体膜融合的 LNP 逃逸[5,6]。 D-Lin-MC3-DMA -LNP 接近中性电荷,zeta 电位为 5±3 mV。虽然 D-Lin-MC3-DMA -LNP 在生理 pH 值下带中性电荷,但已知它们在核内体 pH 值下带正电荷 [7]。 LNPs 通过微流体混合技术合成,由可电离的阳离子脂质 (D-Lin-MC3-DMA)、磷脂、胆固醇和聚乙二醇 (PEG) 以及硫代磷酸化 siRNA 封装的货物组成（50 或 100%）或 mRNA。 LNPs 与生物活性药物 siRNA 形成物理稳定的复合物,持续时间为 94 天[8]。
在小鼠 FVII 模型中,优化组合物 (D-Lin-MC3-DMA) 的 ED50 为 0.005 mg kg 1 siRNA,相对于 40/10/40/10 摩尔比提高了六倍组成[2]。含有 D-Lin-MC3-DMA 的第 2 代 LNP 比含有 DLinDMA 的第 1 代 LNP 强两个数量级以上[4]。