Comparison of DLin-MC3-DMA and ALC-0315 for siRNA Delivery to Hepatocytes and Hepatic Stellate Cells

Abstract

Ionizable cationic lipids are essential for efficient in vivo delivery of RNA by lipid nanoparticles (LNPs). DLin-MC3-DMA (MC3), ALC-0315, and SM-102 are the only ionizable cationic lipids currently clinically approved for RNA therapies. ALC-0315 and SM-102 are structurally similar lipids used in SARS-CoV-2 mRNA vaccines, while MC3 is used in siRNA therapy to knock down transthyretin in hepatocytes. Hepatocytes and hepatic stellate cells (HSCs) are particularly attractive targets for RNA therapy because they synthesize many plasma proteins, including those that influence blood coagulation. While LNPs preferentially accumulate in the liver, evaluating the ability of different ionizable cationic lipids to deliver RNA cargo into distinct cell populations is important for designing RNA−LNP therapies with minimal hepatotoxicity. Here, we directly compared LNPs containing either ALC-0315 or MC3 to knock-down coagulation factor VII (FVII) in hepatocytes and ADAMTS13 in HSCs. At a dose of 1 mg/kg siRNA in mice, LNPs with ALC-0315 achieved a 2-and 10-fold greater knockdown of FVII and ADAMTS13, respectively, compared to LNPs with MC3. At a high dose (5 mg/kg), ALC-0315 LNPs increased markers of liver toxicity (ALT and bile acids), while the same dose of MC3 LNPs did not. These results demonstrate that ALC-0315 LNPs achieves potent siRNAmediated knockdown of target proteins in hepatocytes and HSCs, in mice, though markers of liver toxicity can be observed after a high dose. This study provides an initial comparison that may inform the development of ionizable cationic LNP therapeutics with maximal efficacy and limited toxicity.

Genetically engineered transfusable platelets using mRNA lipid nanoparticles

“…S7), low protein expression and high platelet activation made these undesirable transfection reagents. ALC-0315, which is a potent ionizable LNP in most settings ( 24 , 25 ), gave low expression. Apart from KC2, no ionizable lipid in combination with DSPC achieved higher protein expression than MC3, and only lipids with an acid dissociation constant (pKa) between 6.5 and 6.7 enabled NanoLuc expression when transfected at a pH of 6.5 (fig.…”

Potential of siRNA in COVID-19 therapy: Emphasis on in silico design and nanoparticles based delivery

“…The formed complex can be used to co-deliver mRNA within cationic amphiphilic drugs-assisted LNPs for various applications ( Bogaert et al, 2022 ). The approved LNP-based therapeutic formulations such as Patisiran (Alnylam), Elasomeran (Moderna) use similar ratios (50:10:38.5:1.5) of ILs, helper lipid (DSPC), cholesterol and polyethylene glycol lipids ( Suzuki and Ishihara, 2021 ; Ferraresso et al, 2022 ). The said molar ratio has been reported by many studies as optimum and highly potent ( Carrasco et al, 2021 ; Suzuki et al, 2022 ).…”

Effective Perturbations by Small-Molecule Modulators on Voltage-Dependent Hysteresis of Transmembrane Ionic Currents

“…SM-102 is a synthetic and ionizable amino lipid that has been widely used in combination with other lipids in the formation of lipid nanoparticles [ 50 , 51 , 52 ]. Formulations containing SM-102 have been noticeably used in the development of lipid nanoparticles for the delivery of mRNA-based vaccines.…”