D-Lin-MC3-DMA (Synonyms: MC3)

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.