Table 3. Optimized Lipid Nanoparticle Formulations for LNP Delivery of RNAa.
| ionizable lipid(s) (IL) | optimized formulation (mol %) | cargo, size, and method | outcome | ref |
|---|---|---|---|---|
| DLinDMA | IL (30%)/DSPC (20%)/Chol (48%)/PEG2000-c-DMA (2%) | Luc siRNA 132–182 nm spontaneous vesicle formation by ethanol dilution | 80% knockdown in Neuro 2A cells; uptake not rate-limiting for gene-silencing efficiency | Heyes et al. 2005152 |
| DLin-MC3-DMA | IL (50%)/DSPC (10%)/Chol (38.5%)/PEG2000-DMG (1.5%) | FVII and TTR siRNA 70–90 nm preformed vesicle method | ED50 = 0.005 mg/kg (mouse), ED50 < 0.03 mg/kg (cynomolgus monkeys) | Jayaraman et al. 2012150 |
| DLin-KC2-DMA | IL (57.1%)/DPPC (7.1%)/Chol (34.3%)/PEG-2000-C-DMA (1.4%) | FVII and TTR siRNA 64–85 nm preformed vesicle method and stepwise dilution | in vivo activity achieved at doses of 0.01 mg/kg in mice (C57BL/6) and 0.1 mg/kg in cynomolgus monkeys | Semple et al. 2010153 |
| DLin-MC3-DMA | IL (50%)/DSPC (10%)/Chol (38.5%)/DMPE-PEG-2000 (1.5%) | hEPO mRNA 50–130 nm fast mixing precipitation (microfluidic mixing) | LNPs with 1.5 mol % of DMPE-PEG2000 showed highest hEPO production after 20 h; protein expression in hepatocytes ∼6 hEPO molecules/dosed mRNA, adipocytes ∼0.6 hEPO molecules/dosed mRNA | Yanez Arteta et al. 2018180 |
| DLin-MC3-DMA | IL (50%)/DSPC (10%)/Chol (38.5%)/DMG-PEG2000 (1.5%) | Luc mRNA 83–242 nm microfluidic mixing | albino BALB/c mice (male and female) received subretinal injections of LNPs; LNPs formulated with MC3 and KC2 showed expression that was 2.8- and 3.2-fold higher than DODMA expression; other formulations had significantly lower expression | Patel et al. 2019182 |
| L319 | IL (55%)/DSPC (10%)/Chol (32.5%)/PEG2000-DMG (2.5%) | FVII and TTR mRNA avg. 60 nm spontaneous vesicle formation | ED50 < 0.01 mg/kg (mouse, FVII model), in cynomolgus monkeys ∼70% silencing of TTR mRNA relative to control | Maier et al. 2013134 |
| cKK-E12 | IL (15%)/DOPE (26%)/Chol (40.5%)/SLS (16%)/DMPE-PEG2000 (2.5%) | TRP2 and gp100 mRNA 84–108 nm microfluidic chip | C57BL/6 mice optimized LNPs loaded with TRP2 and gp100 slow tumor growth and extend survival in a B16F10 tumor model | Oberli et al. 2017167 |
| OF-02 | IL (35%)/DOPE (16%)/Chol (46.5%)/DMPE-PEG2000 (2.5%) | hEPO and Luc mRNA 75–112 nm microfluidic mixing | female C57BL/6 mice hEPO expression using OF-02 is double that using cKK-E12; biodistribution (firefly luciferase) is similar to that of cKK-E12 | Fenton et al. 2016169 |
a
Various studies aiming to optimize and characterize LNP formulations for mRNA/siRNA treatments are summarized above. Many of these studies involved screening several lipid species, and only the optimized lipid composition is shown here. Additionally, the RNA cargo, nanoparticle sizes, and LNP formation methods are mentioned. The outcomes of the treatment involving the optimized formulations are reported, measured by various means between studies, and briefly summarized.