DMG-PEG 2000 is a component for the liposome for siRNA delivery, and improved transfection efficiency in vitro.
DMG-PEG 2000 is a component for the liposome for siRNA delivery, and improved transfection efficiency in vitro [1].
DMG-PEG 2000 is the constituent of the nanoliposomes. The nanoparticles are 60 - 100 nm in size and designed for delivery to hepatocytes. The nanoparticles are formed from a mixture of siRNA and four lipid excipients: 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), cholesterol, (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetren-19-yl-4-(dimethylamino) butanoate (DLin-MC3-DMA) and (R)-2,3-bis(tetradecyloxy)propyl 1-(methoxypoly(ethylene glycol)20000)propyl carbamate (DMG-PEG 2000). Both DSPC and cholesterol are well known pharmaceutical ingredients whilst DLin-MC3-DMA and DMG-PEG2000 are novel excipients. Each 1 mL of patisirin contains 3.3 mg DSPC, 6.2 mg cholesterol, 13.0 mg DLin-MC3-DMA and 1.6 mg of DMG-PEG2000 and these lipids associate with the siRNA to form lipid nanoparticles which protects the siRNA from immediate degradation in the circulation and improves delivery to the target site in the liver [2].
参考文献:
[1]. Tianqi Nie, et al. Surface Coating Approach to Overcome Mucosal Entrapment of DNA Nanoparticles for Oral Gene Delivery of Glucagon-like Peptide 1.ACS Appl Mater Interfaces. 2019 Aug 21;11(33):29593-29603.
[2]. Webb C, Forbes N, Roces C B, et al. Using microfluidics for scalable manufacturing of nanomedicines from bench to GMP: A case study using protein-loaded liposomes[J]. International Journal of Pharmaceutics, 2020, 582: 119266.
DMG-PEG 2000 是用于 siRNA 递送的脂质体组分，可提高体外转染效率[1]。
DMG-PEG 2000 是纳米脂质体的成分。纳米粒子的大小为 60-100 nm，专为递送至肝细胞而设计。纳米颗粒由 siRNA 和四种脂质赋形剂的混合物形成:1,2-二硬脂酰-sn-甘油-3-磷酸胆碱 (DSPC)、胆固醇、(6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28 ,31-teren-19-yl-4-(dimethylamino) butanoate (DLin-MC3-DMA) 和 (R)-2,3-bis(tetradecyloxy)propyl 1-(methoxypoly(ethylene glycol)20000)propyl carbamate (DMG -聚乙二醇 2000)。 DSPC 和胆固醇都是众所周知的药物成分，而 DLin-MC3-DMA 和 DMG-PEG2000 是新型赋形剂。每 1 mL patisirin 含有 3.3 mg DSPC、6.2 mg 胆固醇、13.0 mg DLin-MC3-DMA 和 1.6 mg DMG-PEG2000，这些脂质与 siRNA 结合形成脂质纳米颗粒，保护 siRNA 免于在循环中立即降解和改善肝脏中靶点的递送[2]。
| Cell experiment [1]: | |
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Cell lines |
Human hepatocellular carcinoma cell line Hep3B |
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Preparation Method |
TT2-TT8 were formulated with the helper lipid (1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) or 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE)), cholesterol, 1,2-dimyristoyl-sn-glycerol, methoxypolyethylene glycol (DMG-PEG2000) (molar ratio 50/10/38.5/1.5 or based on the orthogonal design table), and FLuc mRNA via pipetting for in vitro studies or via a microfluidic based mixing device for in vivo studies. |
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Reaction Conditions |
Hep3B cells were seeded (2 × 104cells per well) into each well of white 96-well plates in 150 μL of culture medium, allowed to attach overnight in growth medium, and transfected by addition of 20 μL of FLuc mRNA-loaded TT LLNs to growth medium. Transfections were performed in triplicate. After 6 h of transfection, culture medium containing TT LLNs was carefully removed, and 50 μL of serum-free EMEM and 50 μL of Bright-Glo luciferase substrate were mixed and added to each well. |
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Applications |
TT2-TT8 LLNs showed minimal to moderate inhibitory effects on Hep3B cells, and showed a significant positive correlation between transfection efficiency and entrapment efficiency. |
| Animal experiment [2]: | |
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Animal models |
C57BL/6 mice |
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Preparation Method |
C57BL/6 mice were administered intravenously via tail vein injection with free hFIX mRNA, O-TT3 FLuc (an irrelevant mRNA-loaded LLNs as a control), or O-TT3 hFIX at the indicated dosage. Six hours post administration, blood samples were collected and mixed with an anticoagulant solution (3.2% sodium citrate anticoagulant containing 0.17 mg/mL of corn trypsin inhibitor) in a ratio of 9:1, which was then centrifuged for 15 min at 2500 g. hFIX protein level was measured by enzyme-linked immunosorbent (ELISA) assay. |
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Dosage form |
0.55 mg/kg or 1.1 mg/kg |
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Applications |
Wild-type mice produced 1020 ng/mL hFIX at a dose of 0.55 mg/kg and 2057 ng/mL at a dose of 1.1 mg/kg, respectively. No hFIX was detected in the plasma of mice injected with untreated, free hFIX mRNA-, or TT3 LLNs-treated groups |
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参考文献: [1]: Li B, Luo X, Deng B, et al. An orthogonal array optimization of lipid-like nanoparticles for mRNA delivery in vivo[J]. Nano letters, 2015, 15(12): 8099-8107. |
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