Lipid nanoparticles for delivery of nucleic acids and methods of use thereof

Abstract

The present disclosure provides for improved compositions of ionizable lipid nanoparticles for the delivery of therapeutic nucleic acids to cells. Anionic phospholipids, including phosphatidylserine and phosphatidylglycerol are included in the lipid nanoparticles to increase the transfection efficiency in human dendritic cells. The further incorporation of mono-unsaturated alkyl chain analogs in dimethylaminopropyl-dioxolane or heterocyclic ketal ionizable lipids in the formulation demonstrated high levels of transfection in human dendritic cells, compared to other ionizable lipids in the same family, and demonstrated good stability to oxidative damage. Finally, the use of an ammonium salt of phosphatidylserine allows for the efficient production of PS-targeted LNPs.

Description

[0001]Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 LIPID NANOPARTICLES FOR DELIVERY OF NUCLEIC ACIDS AND METHODS OF USE THEREOF RELATED APPLICATIONS This patent application claims the benefit of and priority to U.S. Provisional Patent Application No. 63 / 604,134, filed on November 29, 2023, which is incorporated herein by reference in its entirety. REFERENCE TO SEQUENCE LISTING This specification includes a sequence listing submitted herewith, which includes the file entitled 191016-010901.xml having the following size: 17,585 bytes which was created November 22, 2024, the contents of which are incorporated by reference herein. FIELD The present disclosure relates to cationic ionizable lipids and lipid nanoparticles (LNP). In some embodiments, a LNP comprising one or more cationic ionizable lipid(s) is useful for delivery of a nucleic acid compound, for dendritic cell targeting or methods of using these LNP compositions as vaccines. In some embodiments, a LNP can comprise bioreducible ionizable cationic lipids or unconjugated polyolefinic ionizable cationic lipids. BACKGROUND Lipid nanoparticles (LNP) are used for the delivery of therapeutic nucleic acids to cells. For example, LNP pharmaceutical compositions are employed in vaccines to deliver mRNA therapeutics. LNP formulations typically include an ionizable cationic lipid (ICL). However, it is known in the art that certain ICL compounds are undesirably sensitive to oxidation during storage. Therefore, there is a need for improved ICL compounds with improved stability to oxidative degradation while in storage, while also providing desired transfection activity or potency in cells when incorporated in a LNP with a therapeutic agent such as a nucleic acid. LNP compositions, including stable nucleic acid lipid particle (SNALP) compositions, are useful for delivery of nucleic acid therapies for various infectious diseases. Infectious diseases such as tuberculosis, HIV / AIDS, malaria, and COVID-19 represent significant challenges to human health. Mycobacteria, for example, is a genus of bacteria responsible for tuberculosis (TB). 1 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 According to the World Health Organization, worldwide, TB is one of the top 10 causes of death and the leading cause of death from a single infectious agent. Despite current best efforts, there have been significant challenges in the development of effective vaccines for the prevention of many infectious diseases. New efforts in the identification of individual or combinations of antigenic peptides has helped improved the efficiency of vaccines. Nonetheless, significant opportunities remain in the engineering of adjuvants to help efficiently deliver and present these antigenic sequences to professional antigen presenting cells, like dendritic cells. mRNA coding for antigenic peptides or proteins combined with ionizable cationic lipid nanoparticles represent a particularly promising strategy in the development of a vaccine. There is a need for safe and effective therapies comprising LNP pharmaceutical compositions for delivery of mRNA for treatment and prevention of various diseases, including vaccine compositions. SUMMARY In some embodiments, the present disclosure provides compositions comprising ionizable cationic lipids. Aspects of the disclosure include compositions comprising 3-rac-2,2-di((Z)- octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N-dimethylpropan-1-amine (KC3-OA racemate) or chiral purified forms of the KC3-OA racemate such as KC3-OA(S) and KC3-OA(R), and methods of making and purifying the same. Aspects of the disclosure include compositions comprising 4- rac-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N-dimethylbutan-1-amine (KC4-OA), and methods of making the same. In some embodiments, a composition comprises an ionizable cationic lipid selected from one or more of the following: (a) a racemic mixture of 3-rac-2,2-di((Z)- octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N-dimethylpropan-1-amine (KC3-OA racemate), or KC3-OA enantiomer; and (b) 4-rac-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N- dimethylbutan-1-amine (KC4-OA racemate). In some embodiments, a composition comprises a mixture of (R) and (S) enantiomers of KC3-OA ionizable cationic lipid, or a mixture of (R) and (S) enantiomers of KC4-OA ionizable cationic lipid. In some embodiments, a composition comprises a mixture of (R) and (S) enantiomers of KC3-OA ionizable cationic lipid, or a mixture of (R) and (S) enantiomers of KC4-OA ionizable cationic lipid, and the mixture is racemic. In some embodiments, the disclosure provides certain LNP compositions. In some aspects, the LNP compositions comprise: a nucleic acid; an ionizable cationic lipid at a N / P ratio of 3 to 8 relative to the nucleic acid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; a sterol in a total amount of 0.5-50 mol% of the total lipid content of the LNP 2 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 composition; one or more phospholipids in a total amount of phospholipids of 5-50 mol% of the total lipid content of the LNP composition; and a conjugated lipid in a total amount of 0.5-2.5 mol% of the total lipid content of the LNP composition. In some aspects, the LNP composition is further characterized in that: the nucleic acid is mRNA; the ionizable cationic lipid is present in the LNP composition at a N / P ratio of 4 to 7 relative to the nucleic acid; the sterol is cholesterol; and the conjugated lipid is a PEG-containing conjugated lipid. In some aspects, the one or more phospholipids in the LNP comprise at least two phospholipids having mismatched acyl chain lengths. In some aspects, the one or more phospholipids in the LNP comprise a phosphatidylserine (PS) lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition. In some aspects, the phosphatidylserine (PS) lipid in the LNP consists of, consists essentially of or comprises dipalmitoylphosphatidyl-L-serine ((L-serine)DPPS). In some aspects, the one or more phospholipids in the LNP comprise a phospholipid selected from the group consisting of: distearoylphosphatidylcholine (DSPC) and hydrogenated soy phosphatidylcholine (HSPC). In some aspects, the one or more phospholipids in the LNP consist of distearoylphosphatidylcholine (DSPC) and dipalmitoylphosphatidyl-L-serine ((L-serine)DPPS). In some aspects, the PEG- containing conjugated lipid in the LNP is PEG(2000)-dimyristoylglycerol (PEG-DMG). In some aspects, the LNP composition has 5-50 mol% total phospholipid, including compositions with 5, 10, 15, 20, 25, 30, 35, 40, 45 or 50 mol% total phospholipid. In some embodiments, the LNP composition is further characterized by: the sterol in a total amount of 0.5- 45.5 mol% of the total lipid content of the LNP composition; and the one or more phospholipids in a total amount of phospholipids of 5-50 mol% of the total lipid content of the LNP composition. In some embodiments, the sterol in the LNP composition is cholesterol. In some embodiments, the ionizable cationic lipid is KC3-OA (Racemic). In some embodiments, the ionizable cationic lipid is KC3-OA(S). In some embodiments, the ionizable cationic lipid is KC3-OA(R). In some embodiments, the ionizable cationic lipid is KC4-OA (Racemic). In some embodiments, the ionizable cationic lipid is KC4-OA(S). In some embodiments, the ionizable cationic lipid is KC3- OA(R). In some embodiments, the ionizable cationic lipid is a mixture of KC3-OA and KC4-OA. In some embodiments, the LNP composition comprises a total of 48-54 mol% of the ionizable cationic lipid. In some embodiments, the one or more phospholipids comprise a phosphatidylserine (PS) lipid. In some embodiments, the PS lipid in the LNP composition is DPPS. In some embodiments, 3 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 the PS lipid in the LNP composition is present in a total of 5 mol%. In some embodiments, the LNP composition comprises a DSPC phospholipid. In some embodiments, the LNP composition comprises 7.5-20 mol% of a DSPC phospholipid. In some embodiments, the sterol in the LNP composition is cholesterol. In some embodiments, the LNP composition comprises 25-40 mol% cholesterol. In some embodiments, the sterol in the LNP composition is beta sitosterol. In some embodiments, the LNP composition comprises 33-35.5 mol% beta sitosterol. In some embodiments, the PEG-containing conjugated lipid in the LNP composition is PEG-DMG. In some embodiments, the LNP composition comprises 1.5-4.0 mol% PEG- containing conjugated lipid in the LNP composition is PEG-DMG. In some embodiments, the PEG-containing conjugated lipid in the LNP composition is PEG-DLG. In some embodiments, the LNP composition comprises 1.0-4.0 mol% PEG-containing conjugated lipid in the LNP composition is PEG-DLG. In some embodiments, the composition is a composition selected from Formulations 1-44 in Table 87A. In some embodiments, the composition is a composition selected from Formulations 45-78 in Table 87B. In some embodiments, the composition is a composition selected from Formulations 79-116 in Table 87C. In some embodiments, the composition is a composition selected from Formulations 117-154 in Table 87D. Aspects of the disclosure relate to a lipid nanoparticle (LNP) composition comprising an ionizable cationic lipid selected from one or more of the following (a) a mixture of 3-(S)-2,2- di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N-dimethylpropan-1-amine (KC3-OA(S)) and 3- (R)-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N-dimethylpropan-1-amine (KC3- OA(R)) enantiomer; and (b) 4-(S)-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N- dimethylbutan-1-amine (KC4-OA(S)) and 4-(R)-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4- yl)-N,N-dimethylbutan-1-amine (KC4-OA(R)). In some embodiments, the ionizable cationic lipid is a racemic mixture of 3-rac-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N- dimethylpropan-1-amine (KC3-OA racemate). In other embodiments, the ionizable cationic lipid is a racemic mixture of 4-rac-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N- dimethylbutan-1-amine (KC4-OA racemate). In other embodiments, the ionizable cationic lipid is 3-(R)-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N-dimethylpropan-1-amine (KC3- 4 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 OA(R)). Yet, in other embodiments, the ionizable cationic lipid is 4-(R)-2,2-di((Z)-octadec-9-en- 1-yl)-1,3-dioxolan-4-yl)-N,N-dimethylbutan-1-amine (KC4-OA(R)). In some embodiments, the ionizable cationic lipid is KC3-OA racemate of KC3-OA(S) and KC3-OA(R). In some embodiments, the ionizable cationic lipid is KC3-OA (R) enantiomer. In some embodiments, the ionizable cationic lipid is a KC3-OA enantiomer purified from AKG-KC3- OA racemate of KC3-OA(S) and KC3-OA(R). In some embodiments, the ionizable cationic lipid is KC3-OA mixed enantiomers of KC3-OA(S) and KC3-OA(R), or KC4-OA mixed enantiomers of KC4-OA(S) and KC4-OA(R). In some embodiments, the LNP composition comprises a nucleic acid, the ionizable cationic lipid at a N / P ratio of 3 to 8 relative to the nucleic acid in a total amount of 40-65 mol% of the total lipid content of the LNP composition, a sterol in a total amount of 0.5-50 mol% of the total lipid content of the LNP composition, one or more phospholipids in a total amount of phospholipids of 5-50 mol% of the total lipid content of the LNP composition, and a conjugated lipid in a total amount of 0.5-2.5 mol% of the total lipid content of the LNP composition. In some embodiments, the nucleic acid is mRNA. In some embodiments, the ionizable cationic lipid is present in the LNP composition at a N / P ratio of 4 to 7 relative to the nucleic acid. In some embodiments, the one or more phospholipids comprise a phosphatidylserine (PS) lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition. In some embodiments, the phosphatidylserine (PS) lipid is dipalmitoylphosphatidyl-L-serine ((L-serine)DPPS). In some embodiments, the one or more phospholipids comprise at least two phospholipids having mismatched acyl chain lengths. In some embodiments, the one or more phospholipids comprise a phospholipid selected from the group consisting of: distearoylphosphatidylcholine (DSPC), hydrogenated soy phosphatidylcholine (HSPC), and dipalmitoylphosphatidyl-L-serine ((L- serine)DPPS). In some embodiments, the sterol is cholesterol. In other embodiments, the sterol is beta- sitosterol. In some embodiments, the conjugated lipid is a PEG-containing conjugated lipid. In some embodiments, the PEG-containing conjugated lipid is selected from the group consisting of: PEG(2000)-dimyristoylglycerol (PEG-DMG), 1,2-dilauroyl-sn-glycero-3-phosphoethanolamine- N-[methoxy(polyethylene glycol)-2000] (PEG-DLPE), and PEG(2000)-dilauroylglycerol (PEG- DLG). 5 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 In some embodiments, the LNP composition comprises 25-40 mol% cholesterol. In some embodiments, the LNP composition comprises 1.5-4.0 mol% of the PEG- containing conjugated lipid. In some embodiments, the one or more phospholipids comprise distearoylphosphatidylcholine (DSPC). In some embodiments, the PEG-containing conjugated lipid is PEG(2000)- dimyristoylglycerol (PEG-DMG) or PEG(2000)-dilauroylglycerol (PEG-DLG). Aspects of the disclosure relates to a vaccine comprising the LNP composition provided herein. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a depiction of the oxidative degradation mechanisms of lipid esters of linoleic acid containing conjugated multiple unsaturations that are particularly sensitive to oxidation. FIG.2 shows the reaction of the reduced c-terminal cysteine of a Fab’ antibody fragment with a maleimide terminated-poly(ethylene glycol) 2000 derivatized distearoylphosphatidylethanolamine. R1 and R2 are stearic acid. The final antibody lipopolymer conjugate is an intermediate that is subsequently inserted into the outer lipid layer of lipidic nanoparticle to make it actively targeted. FIG. 3A. Impact of DSPS inclusion from 0-2.5 mol % on transfection efficiency of dendritic cells (MutuDC1940) using mCherry mRNA LNPs formulated with DLin-KC2-DMA as the ionizable cationic lipid. ICL was kept at 50 mol%, cholesterol at 38.5 mol%, PEG-DMG at 1.5 mol% and the DSPS content varied. Inclusion of DSPS was made by reducing the DSPC content by the same mol% of DSPS that was added. Cells were incubated with each formulation at a concentration of 1 ug mRNA / mL for 24 h. UT sample corresponds to cells where no LNPs were added. Lipofect refers to Lipofectamine treated sample. FIG. 3B. Impact of DSPS inclusion from 0-7.5 mol % on transfection efficiency of dendritic cells (MutuDC1940) using mCherry mRNA LNPs formulated with DLin-KC2-DMA as the ionizable cationic lipid. ICL was kept at 50 mol%, cholesterol at 38.5 mol%, PEG-DMG at 1.5 mol% and the DSPS content varied. Inclusion of DSPS was made by reducing the DSPC content by the same mol% of DSPS that was added. Cells were incubated with each formulation at a concentration of 1 ug mRNA / mL for 24 h. UT sample corresponds to cells where no LNPs were added. Lipofect refers to Lipofectamine treated sample. 6 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 FIG. 3C. Impact of DSPS inclusion from 0-7.5 mol % on transfection efficiency of dendritic cells (MutuDC1940) using mCherry mRNA LNPs formulated with DLin-KC2-DMA as the ionizable cationic lipid. ICL was kept at 50 mol%, cholesterol at 38.5 mol%, PEG-DMG at 1.5 mol% and the DSPS content varied. Inclusion of DSPS was made by reducing the DSPC content by the same mol% of DSPS that was added. Cells were incubated with each formulation at a concentration of 0.3 ug mRNA / mL for 24 h. UT sample corresponds to where no LNPs were added. FIG. 3D. Impact of DSPS inclusion from 0-7.5 mol % on transfection efficiency of dendritic cells (MutuDC1940) using mCherry mRNA LNPs formulated with DLin-KC2-DMA as the ionizable cationic lipid. ICL was kept at 50 mol%, cholesterol at 38.5 mol%, PEG-DMG at 1.5 mol% and the DSPS content varied. Inclusion of DSPS was made by reducing the DSPC content by the same mol% of DSPS that was added Cells were incubated with each formulation at a concentration of 0.1 ug mRNA / mL for 24 h. UT sample corresponds to cells where no LNPs were added. FIG. 4. Transfection of murine dendritic cells (MutuDC1940) using LNPs containing various ICLs (KC2, KC2-OA, KC3-OA, and SM-102) and 5 mol % DSPS, and comparison to LNPs using Glu-DSPE or Suc-DSPE rather than DSPS. UT sample corresponds to cells where no LNPs were added. FIG.5. DSPS or DPPS increase mCherry LNP transfection with KC2, KC2-01, KC2-PA, KC3-01, and KC3-OA comprising ICLs. UT sample corresponds to cells where no LNPs were added. FIG.6A. Comparison of various chemical forms of phosphatidylserine with AKG-UO-1 containing LNPs in transfecting murine dendritic cells. UT sample corresponds to cells where no LNPs were added. Lipo refers to Lipofectamine MessengerMax (ThermoFisher) used according to manufacturer’s instructions at the same dosage level as the LNPs. FIG.6B. Comparison of DSPS to other negatively charged phospholipids in transfecting murine dendritic cells using AKG-UO1 containing LNPs. UT sample corresponds to cells where no LNPs were added. Lipo refers to Lipofectamine MessengerMax (ThermoFisher) used according to manufacturer’s instructions at the same dosage level as the LNPs. FIG.7. Impact of DSPS concentration in AUG-UO-1 containing LNPs on transfection of dendritic cells. UT sample corresponds to cells where no LNPs were added. 7 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 FIG. 8. Impact of PEG-DMG concentration in AUG-UO-1 containing LNPs with and without 5 mol % DSPS on transfection of dendritic cells. The Y-axis shows the % PEG used in the composition followed by the concentration of mRNA added to the cells (0.11, 0.33, or 1 µg / mL). UT sample corresponds to cells where no LNPs were added. FIG. 9A. Oxidative degradation of lipid suspensions of ICLs with a single methylene between two olefins (KC2, KC3, and O-11769) and those with four methylenes between the two olefins (KC2-01, KC3-01, and UO-1). FIG. 9B. Oxidative degradation of liposomes containing O-11769, an ICL with a single methylene between two olefins liposomes containing UO-1, an ICL with four methylenes between the two olefins. FIG.10A. Effect of N / P on mCherry expression of KC2-01 containing LNPs at 1 µg / ml in murine dendritic cells. UT sample corresponds to cells where no LNPs were added. FIG.10B. Effect of N / P on mCherry expression of KC2-01 containing LNPs at 0.33 µg / ml in murine dendritic cells. UT sample corresponds to cells where no LNPs were added. FIG. 11. Transfection efficiency of LNPs with and without DSPS (7.5 mol %) and containing different ionizable cationic lipids. UT sample corresponds to cells where no LNPs were added. FIG. 12. Transfection efficiency of LNP formulations containing various concentrations of DOPS (0, 10, and 25 mol % as % of total lipid) and mCherry mRNA in murine dendritic cells. FIG. 13A. mRNA sequence of VRN-029, a SARS-COV2 spike protein generating sequence. FIG. 13B. The effect of PEG-DMG (C14) concentration (mol %) on LNP vaccine immunogenicity. Total anti-spike antibody titers and CD4 responses from mice immunized with mRNA-LNPs using 7.5% DSPS and the ionizable lipid UO1 with increasing mol% of PEG-DMG. The middle graph shows day 34 endpoint antibody titers. The right graph shows the corresponding CD4 T cell responses. FIG. 13C. The effect of PEG-DPPE (C16) concentration (mol %) on LNP vaccine immunogenicity. Total anti-spike antibody titers from mice immunized with mRNA-LNPs using 7.5% DSPS and the ionizable lipid UO1 with increasing mol% of PEG-DPPE. The middle graph shows day 34 endpoint antibody titers. The mol% of PEG-DPPE inversely impacted antibody levels. The right graph shows the corresponding CD4 T cell responses. 8 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 FIG.13D. Total anti-spike antibody titers and CD4 responses from mice immunized with mRNA-LNPs using 7.5% DSPS and the ionizable lipid KC2OA with either 1.5 mol% PEG-DMG (14C) or PEG-DSG (18C). The left graph shows day 34 endpoint antibody titers. The right graph shows the corresponding CD4 T cell responses. FIG.13E. Total anti-spike antibody titers and CD4 responses from mice immunized with mRNA-LNPs using 7.5% DSPS and the ionizable lipid UO1 with either 1.5 mol% PEG-DMG (14C) or PEG-DSG (18C). The left graph shows day 34 endpoint antibody titers. The right graph shows the corresponding CD4 T cell responses. FIG. 13F. Effect of phosphatidylserine incorporation in mRNA-LNP immunogenicity. Total anti-spike antibody titers (A) and spike-specific CD4 T cell responses from mice immunized with mRNA-LNPs using various ionizable lipids and PEG-lipids plus / minus 7.5 mol% DSPS Antibody data were log-transformed and analyzed using two-way ANOVA with a Sidak’s multiple comparison test. CD4 T cell data were analyzed using a REML mixed-effects model with a Sidak’s multiple comparison test. FIG.13G. Effect of phosphatidylserine lipid tail (DPPS vs DSPS) composition on mRNA- LNP priming of B (Panel A) and T cell (Panel B) responses. Antibody data were log-transformed prior to analysis. Data were analyzed using one-way ANOVA with a Tukey’s multiple comparison test. FIG.14A. Comparison of the mCherry expression of KC2-01 LNPs, 7.5 mol% DSPS (D isomer) and DSPS (L isomer) at 1 µg / mL mRNA for 24h. FIG.14B. Comparison of the mCherry expression of KC2-01 LNPs, 7.5 mol% DSPS (D isomer) and DSPS (L isomer) at 0.33 µg / mL mRNA for 24h. FIG.15. Comparison of the mCherry expression of KC2 LNPs, with 5 and 7.5 mol% DSPS (L-isomer) to LNPs prepared with SM-102 or ALC-0315 at 1 µg / mL mRNA for 24h. The Y-axis is mean fluorescence intensity (MFI). UT sample corresponds to cells where no LNPs were added. FIG. 16. Comparison of the mCherry expression of UO1, UO6 and UO7 formulations alone, or with added 7.5mol% D-isomer of DSPS, at 1 µg / mL mRNA for 24 h. UT sample corresponds to cells where no LNPs were added. FIG.17. Comparison of the mCherry expression of UO1, SM102, ALC-0315 formulations alone, or with added DSPS, at 1 µg / mL mRNA for 24h. Lipo refers to Lipofectamine 9 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 MessengerMax (ThermoFisher) used according to manufacturer’s instructions at the same dosage level as the LNPs. UT sample corresponds to cells where no LNPs were added. FIG. 18. Oxidative degradation of liposomes containing KC3 (DLin-KC3-DMA), a polyunsaturated ICL with a single methylene between two olefins, to liposomes containing ICLs with monounsaturated alkyl chains (KC3-OA, KC3-PA, or KC3-C17(C8:1)) and the fully saturated ICL, KC3-C17. Effect of hydrogen peroxide on the stability of individual ionizable cationic lipids measured by CAD-HPLC. FIG. 19. Comparison of the mCherry expression of UO1, UO1A, and KC3-OA LNP formulations alone, or with added DSPS, at 0.1 and 1 µg / mL mRNA for 24h in human dendritic cells. Untreated DC sample corresponds to human dendritic cells where no LNPs were added. FIG. 20. Comparison of the mCherry expression in murine dendritic cells of LNPs containing the polyunsaturated KC3, the monounsaturated KC3-OA, KC3-PA, or KC3C17(C8:1), and the fully saturated KC3C17, all with or without DPPS (NH4+salt), at 0.3 or 1 µg / mL mRNA for 24h. UT sample corresponds to cells where no LNPs were added. FIG. 21. Comparison of the mCherry expression in human dendritic cells of LNPs containing the polyunsaturated KC2 or KC3 with a single methylene between two olefins, polyunsaturated KC3-01 with four methylenes between two olefins, monounsaturated KC3-OA, KC3-PA, or KC3C17(C8:1), and ALC-0315, all with except ALC-0315 with DPPS (NH4+salt), at 0.1 or 1 µg / mL mRNA for 24h. Untreated samples correspond to cells where no LNPs were added. FIG.22. Comparison of the mCherry expression of LNP formulations with 5 mol % DSPS and 46-54 mol % of KC3-OA to ALC-0315 and SM-102 LNP controls, at 0.1 and 1 µg / mL mRNA for 24h in human dendritic cells. Untreated DC sample corresponds to human dendritic cells where no LNPs were added. FIG.23. Comparison of the mCherry expression of LNP formulations with 0 or 5 mol % DSPS and 50 mol % KC2-O1 at N / P ratios of 4-7, at 0.1 µg / mL mRNA for 24h in human dendritic cells. These were also compared to LNPs containing KC3-OA and 5 mol % DSPS at N / P of 5. Untreated DC sample corresponds to human dendritic cells where no LNPs were added. FIG.24A. Comparison of polyunsaturated KC3 with monounsaturated KC3-OA and KC3- PA containing LNP formulations on vaccine immunogenicity. Total anti-spike antibody titers from mice immunized with mRNA-LNPs using 5 mol % DSPS or DPPS-targeted LNPs containing 10 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 either KC3, KC3-OA, or KC3-PA. For KC3-OA and KC3-PA LNPs, each formulation was also evaluated with either the C16 DPPC or C18 DSPC neutral phosphatidylcholine component. All LNPs contained 1.5 mol % of PEG-DMG. The graph shows day 21 endpoint antibody titers after the initial prime injection of 1 µg mRNA per mouse. FIG.24B. Comparison of polyunsaturated KC3 with monounsaturated KC3-OA and KC3- PA containing LNP formulations on vaccine immunogenicity. Total anti-spike antibody titers from mice immunized with mRNA-LNPs using 5 mol % DSPS or DPPS-targeted LNPs containing either KC3, KC3-OA, or KC3-PA. For KC3-OA and KC3-PA LNPs, each formulation was also evaluated with either the C16 DPPC or C18 DSPC neutral phosphatidylcholine component. All LNPs contained 1.5 mol % of PEG-DMG. The graph shows day 34 endpoint antibody titers after the prime then boost on day 21 of 1 µg mRNA per mouse. FIG. 25A. Comparison of the mCherry expression of LNP formulations with 5 mol % DSPS and 43-48 mol % of KC3-OA to ALC-0315 and SM-102 LNP controls, at 1 µg / mL mRNA for 24h in human dendritic cells KC3-OA LNPs prepared at 45 mol % KC3-OA and 5 mol % DSPS of total lipid were also compared at N / P ratios of 5, 5.5, 6.0, and 6.5. Finally, LNPs with 45 mol % KC3-OA at N / P of 5 and 6 were evaluated with PEG-SA, at either 1 or 3 mol %, in place of 1.5 mol % PEG-DMG. Untreated DC sample corresponds to human dendritic cells where no LNPs were added. FIG. 25B. Comparison of the mCherry expression of LNP formulations with 5 mol % DSPS and 43-48 mol % of KC3-OA to ALC-0315 and SM-102 LNP controls, at 0.1 µg / mL mRNA for 24h in human dendritic cells KC3-OA LNPs prepared at 45 mol % KC3-OA and 5 mol % DSPS of total lipid were also compared at N / P ratios of 5, 5.5, 6.0, and 6.5. Finally, LNPs with 45 mol % KC3-OA at N / P of 5 and 6 were evaluated with PEG-SA, at either 1 or 3 mol %, in place of 1.5 mol % PEG-DMG. Untreated DC sample corresponds to human dendritic cells where no LNPs were added. FIG. 26A. Comparison of the mCherry expression of 50 mol % UO-1 containing LNP formulations with 7.5 mol % of various anionic phospholipids in human dendritic cells following incubation for 24 h at 1 µg / mL mRNA. All LNPs included 2.5 mol % of DSPC, 50 mol % of UO- 1, and 1.5 mol % of PEG-DMG. The anionic phospholipids included the phosphatidylserines, DOPS, DSPS, DPPS, and DMPS, distearoylphosphatidylglycerol (DSPG), and N-glutaryl- distearoylphophatidylethanolamine (Glu-DSPE) and N-succinyl-distearoylphophatidylethanol- 11 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 amine (Suc-DSPE) and were included at 7.5 mol %, except for DSPG was compared at both 5 and 7.5 mol %. FIG. 26B. Comparison of the mCherry expression of 50 mol % UO-1 containing LNP formulations with 7.5 mol % of various anionic phospholipids in human dendritic cells following incubation for 24 h at 0.1 µg / mL mRNA. All LNPs included 2.5 mol % of DSPC, 50 mol % of UO-1, and 1.5 mol % of PEG-DMG. The anionic phospholipids included the phosphatidylserines, DOPS, DSPS, DPPS, and DMPS, distearoylphosphatidylglycerol (DSPG), and N-glutaryl- distearoylphophatidylethanolamine (Glu-DSPE) and N-succinyl-distearoylphophatidylethanol- amine (Suc-DSPE) and were included at 7.5 mol %, except for DSPG was compared at both 5 and 7.5 mol %. FIG. 27A. Comparison of the mCherry expression of UO-1 or KC3-01 containing LNP formulations with 0-10 mol % of DSPG in human dendritic cells following incubation for 24 h at 1 µg / mL mRNA. ALC-0315 and SM-102 LNPs controls were also included at 1 µg / mL mRNA and untreated DC sample corresponds to human dendritic cells where no LNPs were added. FIG. 27B. Comparison of the mCherry expression of UO-1 or KC3-01 containing LNP formulations with 0-10 mol % of DSPG in human dendritic cells following incubation for 24 h at 0.1 µg / mL mRNA. ALC-0315 and SM-102 LNPs controls were also included at 0.1 µg / mL mRNA and untreated DC sample corresponds to human dendritic cells where no LNPs were added. FIG. 28. Comparison of the mCherry expression in murine dendritic cells of LNPs containing UO1, UO6, or UO7 with 7.5 mol % DSPS after incubation at 1 µg / mL mRNA for 24h. UT sample corresponds to cells where no LNPs were added. FIG.29. Comparison of dilinoleyl KC2, monounsaturated KC3-OA, and four methylene interrupted poly unsaturated ICLs (KC3-01, AKG-UO1, and AKG-UO9) containing LNP formulations on vaccine immunogenicity. ALC-0315 containing LNPs were included as a control. All LNPs contained 1.5 mol % of PEG-DMG. Total anti-spike antibody titers from mice immunized with mRNA-LNPs were determined on day 21 after the initial prime injection of 1 µg mRNA per mouse on day 1. FIG.30A. Comparison of the mCherry expression of 48 mol % KC3-OA containing LNP formulations with 5 mol % of various anionic phospholipids in human dendritic cells following incubation for 24 h at 1 µg / mL mRNA. All LNPs included 2.5 mol % of DSPC, 50 mol % of UO- 1, and 1.5 mol % of PEG-DMG. The anionic phospholipids included the phosphatidylglycerols, 12 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 DOPG, DSPG, DPPG, and DMPG, as well as DSPS. In some LNPs, the DSPG and DSPS were combined either alone or together with DSPC. Two donors were used to produce human dendritic cells in this study and untreated DC sample corresponds to human dendritic cells where no LNPs were added. FIG.30B. Comparison of the mCherry expression of 48 mol % KC3-OA containing LNP formulations with 5 mol % of various anionic phospholipids in human dendritic cells following incubation for 24 h at 0.1 µg / mL mRNA. All LNPs included 2.5 mol % of DSPC, 50 mol % of UO-1, and 1.5 mol % of PEG-DMG. The anionic phospholipids included the phosphatidylglycerols, DOPG, DSPG, DPPG, and DMPG, as well as DSPS. In some LNPs, the DSPG and DSPS were combined either alone or together with DSPC. Two donors were used to produce human dendritic cells in this study and untreated DC sample corresponds to human dendritic cells where no LNPs were added. FIG. 31. Comparison of the mCherry expression in murine dendritic cells of LNPs containing KC3-OA LNPs with either 5 mol % DSPS (Na+salt) or 5 mol % DPPS (NH4+salt) after incubation at 1 µg / mL mRNA for 24h. ALC-0315 and SM-102 LNPs controls were also included at 1 µg / mL mRNA. UT sample corresponds to cells where no LNPs were added. FIG.32. Graph of LNP uptake over time in MutDC1940 cells incubated with various LNP formulations of mCherry mRNA (0.2 µg / mL) labeled with a fluorescent lipid label DiIC18(5)-DS (DiI5-DS) (Example 47). At indicated times LNP uptake was quantified by flow cytometry of the label (RL-1 fluorescence channel) and plotted as median cell fluorescence intensity. Data are the average of triplicate runs. Error bars, standard deviation. Formulations: AKG + PS, KC3OA+PS; AKG – PS, KC3OA-PS; SM102; ALC-0315. Untreated refers to the cells without LNP treatment. FIG.33. Graph of LNP mRNA expression over time in MutuDC1940 cells incubated with various LNP formulations of mCherry mRNA (0.2 µg / mL) labeled with a fluorescent lipid label DiIC18(5)-DS (DiI5-DS) (Example 47). At indicated times mCherry protein expression was quantified by flow cytometry by the protein fluorescence (YL-2 fluorescence channel) and plotted as median cell fluorescence intensity. Data are the average of triplicate runs. Error bars, standard deviation. Formulations: AKG + PS, KC3OA+PS; AKG – PS, KC3OA-PS; SM102; ALC-0315. Untreated refers to the cells without LNP treatment. FIG.34. The effect of cell pre-treatment with “blocking liposomes” at 10x the LNP lipid concentration on the uptake of DiI5-DS-labeled LNPs by MutuDC1940 cells (Example 48). The 13 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 liposomes were added 15 min prior to the LNPs. After 1h of LNP exposure the uptake of LNPs was measured as DiI5-DS fluorescence by flow cytometry (RL1-A channel) and plotted as median cellular fluorescence intensity. Data are the average of quadruplicate runs. Error bars, standard deviation. LNP type is shown at the horizontal axis. The legend indicates the amount and nature of the PS component in the “blocking liposomes” The “%” indicates mol% of PS in the blocking liposome formulation related to the total lipid. FIG. 35. The effect of PS targeting agents on mCherry expression in MutuDC1940 cell after 3 hours incubation with mCherry mRNA-LNP constructs (0.3 µg / mL mRNA) prepared with various ICL (Example 49). The expression was quantified by flow cytometry by the mCherry protein fluorescence (YL2-A fluorescence channel) and plotted as median cell fluorescence intensity. Data are the average of quadruplicate runs. Error bars, standard deviation. ICL: UO1, KC2, KC3OA. Targeting PS: DPPS, DSPS, D-DSPS, no targeting (NT). UT refers to the cells without LNP treatment. FIG. 36. The effect of PS targeting agents on mCherry expression in MutuDC1940 cell after 24 hours incubation with mCherry mRNA-LNP constructs (0.3 µg / mL mRNA) prepared with various ICL (Example 49). The expression was quantified by flow cytometry by the mCherry protein fluorescence (YL2-A fluorescence channel) and plotted as median cell fluorescence intensity. Data are the average of quadruplicate runs. Error bars, standard deviation. ICL: UO1, KC2, KC3OA. Targeting PS: DPPS, DSPS, D-DSPS, no targeting (NT). UT refers to the cells without LNP treatment. FIG.37. The effect of PS targeting agents on the LNP uptake in MutuDC1940 cell after 3 hours incubation with mCherry mRNA-LNP constructs (0.3 µg / mL mRNA) prepared with various ICL (Example 49). The expression was quantified by flow cytometry by the DiI5-DS (DiI) lipid label fluorescence (RL1A fluorescence channel) and plotted as median cell fluorescence intensity. Data are the average of quadruplicate runs. Error bars, standard deviation. ICL: UO1, KC2, KC3OA. Targeting PS: DPPS, DSPS, D-DSPS, no targeting (NT). UT refers to the cells without LNP treatment. FIG.38. The effect of PS targeting agents on the LNP uptake in MutuDC1940 cell after 24 hours incubation with mCherry mRNA-LNP constructs (0.3 µg / mL mRNA) prepared with various ICL (Example 49). The expression was quantified by flow cytometry by the DiI5-DS (DiI) lipid label fluorescence (RL1A fluorescence channel) and plotted as median cell fluorescence 14 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 intensity. Data are the average of quadruplicate runs. Error bars, standard deviation. ICL: UO1, KC2, KC3OA. Targeting PS: DPPS, DSPS, D-DSPS, no targeting (NT). UT refers to the cells without LNP treatment. FIG. 39. Expression of mCherry mRNA delivered to MutuDC1940 cells by LNP formulations of mCherry mRNA (0.3 µg / mL) formulated in KC3OA / DPPS LNPs prepared with PEG-DMG and various amounts of PEG-DLG (Example 50). The expression was quantified by flow cytometry of mCherry fluorescence 24 hours after transfection with LNPs at 1.0 and 0.1 μg / mL mRNA. PBMCs were taken from two donors and each was run in triplicate. The bars are average across the combined donor replicate data. LNP compositions are indicated at the X-axis. “%” refers to the mol% of the PEG-lipid component relative to the total LNP lipid. FIG. 40. Anti-SARS-COV-2 spike protein antibody titers in the sera of Balb / C mice immunized with the VRN029 spike mRNA formulated in KC3-PA / DSPC / DPPS-NH4 / Chol / PEG- lipid LNPs containing PRG-DMG or PEG-DLG (N=5) (Example 51). The sera were collected 2 weeks post boost dose (day 35 post prime dose) of 1 μg mRNA / mouse. The titers were determined using 4x background as an endpoint of ELISA test. The bars represent geometric mean titers across the groups, the error intervals are standard deviations of the log transformed data, the markers show individual animal titers. The numbers at the PEG-lipid designation are mol% of the PEG- lipid relative to the LNP total lipid. The dotted line shows the upper limit of detection in the titer assay. FIG. 41A. Anti-SARS-COV-2 spike protein antibody titers in the sera of Balb / C mice immunized with the VRN029 spike mRNA formulated in KC3-OA / DSPC / DPPS-NH4 / Chol / PEG- lipid LNPs containing PEG-DMG or PEG-DLG (N=5) (Example 52). The sera were collected 3 weeks post prime mRNA-LNP dose of 1 μg mRNA / mouse. The titers were determined using 4x background as an endpoint of ELISA test. The bars represent geometric mean titers across the groups, the error intervals are standard deviations of the log transformed data, the markers show individual animal titers. The difference between the groups is statistically significant. The dotted line shows the lower limit of detection in the titer assay. FIG. 41B. Anti-SARS-COV-2 spike protein antibody titers in the sera of Balb / C mice immunized with the VRN029 spike mRNA formulated in KC3-OA / DSPC / DPPS-NH4 / Chol / PEG- lipid LNPs containing various amounts of DPPS and either PEG-DMG or PEG-DLG (N=5) (Example 52). The sera were collected 2 weeks after the boost mRNA-LNP dose of 1 μg 15 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 mRNA / mouse administered 3 weeks after the prime. The titers were determined using 4x background as an endpoint of ELISA test. The bars represent geometric mean titers across the groups, the error intervals are standard deviations of the log transformed data, the markers show individual animal titers The dotted line shows the lower limit of detection in the titer assay. “%” signifies the mol% of the lipid component relative to the total LNP lipid. FIG.42. Expression of mCherry mRNA delivered to human PBMC-derived dendritic cells by KC3OA / DSPC / DPPS-NH4 / Chol / PEG-lipid LNPs prepared with various amounts of PEG- DMG or PEG-DLPE (Example 53). The expression was quantified by flow cytometry of mCherry fluorescence 24 hours after transfection with LNPs at 0.3 μg / mL mRNA. The bars show average median cell fluorescence intensity of the replicates. Error intervals are standard deviation. Mol% PEG-lipid refers to the mol% relative to the total LNP lipid. FIG. 43A. Anti-SARS-COV-2 spike protein antibody titers in the sera of Balb / C mice immunized with the VRN029 spike mRNA formulated in KC3-OA / DSPC / DPPS-NH4 / Chol / PEG- DMG LNPs containing various amounts of DPPS (N=5) (Example 54). The sera were collected on Day 20 after the prime mRNA dose of 0.3 μg mRNA / mouse. The titers were determined using 4x background as an endpoint of ELISA test. The bars represent geometric mean titers across the groups, the error intervals are standard deviations of the log transformed data, the markers show individual animal titers. The asterisk denotes statistical significance of the difference between the groups at p=0.05. The groups with p value of 0.059 are also shown. The dotted line shows the lower limit of detection in the titer assay. The numbers at the LNP lipid composition labels are mol% of respective components relative to the total LNP lipid. FIG. 43B. Anti-SARS-COV-2 spike protein antibody titers in the sera of Balb / C mice immunized with the VRN029 spike mRNA formulated in KC3-OA / DSPC / DPPS-NH4 / Chol / PEG- lipid LNPs containing various amounts of PEG-DMG or PEG-DLG, and of the ALC-0315-based nontargeted LNP formulation (N=5) (Example 54). The sera were collected at 2 weeks after the boost injection made at 3 weeks following the prime mRNA-LNP dose of 0.3 μg mRNA / mouse. The titers were determined using 4x background as an endpoint of ELISA test. The bars represent geometric mean titers across the groups, the error intervals are standard deviations of the log transformed data, the markers show individual animal titers. The dotted line shows the lower limit of detection in the titer assay. The numbers at the LNP lipid composition labels are mol% of respective components relative to the total LNP lipid. 16 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 FIG. 43C. Anti-SARS-COV-2 spike protein antibody titers in the sera of Balb / C mice immunized with the VRN029 spike mRNA formulated in KC3-OA / DSPC / PS / Chol / PEG-lipid LNPs containing DSPS-Na or DPPS-NH4 as a PS component at 5 mol% relative to the total LNP lipid, and PEG-DMG or PEG-DLG as a PEG-lipid component at 1.5 mol% relative to the total LNP lipid (N=5) (Example 54). The sera were collected at 2 weeks after the boost injection made at 3 weeks following the prime mRNA-LNP dose of 0.3 μg mRNA / mouse (Day 35 post prime). The titers were determined using 4x background as an endpoint of ELISA test. The bars represent geometric mean titers across the groups, the error intervals are standard deviations of the log transformed data, the markers show individual animal titers. The dotted line shows the lower limit of detection in the titer assay. The numbers at the LNP lipid composition labels are mol% of PEG- lipid relative to the total LNP lipid. FIG. 44. Anti-SARS-COV-2 spike protein antibody titers in the sera of Balb / C mice immunized with various SARS-COV-2 spike mRNA (VRN029, VRN118, VRN119) formulated in KC3-OA / DSPC / DPPS-NH4 / Chol / PEG-DMG LNPs or ALC-0315 LNPs (N=5) (Example 55). The sera were collected on Day 20 after the prime mRNA-LNP dose of 0.3 μg mRNA / mouse. The titers were determined using 4x background as an endpoint of ELISA test. The bars represent geometric mean titers across the groups, the error intervals are standard deviations of the log transformed data, the markers show individual animal titers. The dotted line shows the lower limit of detection in the titer assay. FIG. 45A. Anti-SARS-COV-2 spike protein antibody titers in the sera of Balb / C mice immunized with the VRN029 spike mRNA formulated in ALC-0315-based LNP, SM-102-based LNP, or KC3-OA / DSPC / DPPS-NH4 / Chol / PEG-DMG LNPs (N=5) (Example 56). The sera were collected at 2 weeks after the boost injection made at 3 weeks following the prime mRNA-LNP dose of 0.1, 0.3, or 1.0 μg mRNA / mouse (Day 35 post prime). The titers were determined using 4x background as an endpoint of ELISA test. The bars represent geometric mean titers across the groups, the error intervals are standard deviations of the log transformed data, the markers show individual animal titers. The dotted line shows the lower limit of detection in the titer assay. The numbers at the LNP lipid composition labels are mol% of PEG-lipid relative to the total LNP lipid. FIG. 45B. Anti-SARS-COV-2 spike protein antibody titers in the sera of Balb / C mice immunized with the VRN029 spike mRNA formulated in KC3-OA / DSPC / DPPS-NH4 / Chol / PEG- DMG LNPs with different amounts of PEG-DMG and administered at different doses (N=5) 17 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 (Example 56). The sera were collected at 2 weeks after the boost injection made at 3 weeks following the prime mRNA-LNP dose of 0.1, 0.3, or 1.0 μg mRNA / mouse (Day 35 post prime). The titers were determined using 4x background as an endpoint of ELISA test. The bars represent geometric mean titers across the groups, the error intervals are standard deviations of the log transformed data, the markers show individual animal titers. The dotted line shows the lower limit of detection in the titer assay. The numbers at the LNP lipid composition labels are mol% of the corresponding lipid components relative to the total LNP lipid. FIG. 46A. Anti-SARS-COV-2 spike protein antibody titers in the sera of Balb / C mice immunized with the VRN029 spike mRNA formulated in KC3-OA / DSPC / DPPS-NH4 / Chol / PEG- DMG (targeted KC3-OA) LNPs , KC3-OA / DSPC / Chol / PEG-DMG (non-targeted, KC3-OA) LNPs, or ALC-0315 LNPs (Example 57). The sera were collected at Day 20 following the prime mRNA-LNP dose of 0.1 μg mRNA / mouse. The titers were determined using 4x background as an endpoint of ELISA test. The bars represent geometric mean titers across the groups, the error intervals are standard deviations of the log transformed data, the markers show individual animal titers. The dotted line shows the lower limit of detection in the titer assay. FIG. 46B. Anti-SARS-COV-2 spike protein antibody titers in the sera of Balb / C mice immunized with the VRN029 spike mRNA formulated in KC3-OA / DSPC / DPPS-NH4 / Chol / PEG- DMG (targeted KC3-OA) LNPs, KC3-OA / DSPC / Chol / PEG-DMG (non-targeted, KC3-OA) LNPs, or ALC-0315 LNPs (Example 57). The sera were collected at 2 weeks after the boost injection made at 3 weeks following the prime mRNA-LNP dose of 0.1 μg mRNA / mouse (Day 35 post prime). The titers were determined using 4x background as an endpoint of ELISA test. The bars represent geometric mean titers across the groups, the error intervals are standard deviations of the log transformed data, the markers show individual animal titers. The dotted line shows the lower limit of detection in the titer assay. FIG.47A and FIG.47B. Synthetic schemes for UO-1 series of ionizable cationic lipids. FIG.48. Synthetic scheme for KC2-01 and KC3-01 ionizable cationic lipids. FIG. 49. Synthetic scheme for monounsaturated KC2-0A, KC2-PA, and KC3-OA ionizable cationic lipids. FIG. 50. Synthetic scheme for monounsaturated KC3-C17(C8:1) and fully unsaturated KC3-C17 ionizable cationic lipids. FIG.51 Synthetic scheme for distearoylphophatidyl-D-serine. 18 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 DETAILED DESCRIPTION It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the compositions and methods of the present disclosure. Liposomal nanoparticle (LNP) compositions can comprise an ionizable lipid, a sterol, and one or more phospholipids. In some embodiments, the LNP compositions further comprise a nucleic acid such as mRNA for administration in a pharmaceutical composition such as a vaccine. In some embodiments, the LNP compositions optionally further comprise a conjugated lipid. Lipid Nanoparticle (LNP) compositions comprising mRNA include Stabilized Nucleic Acid Lipid Particles (SNALP) used as a vehicle for the systemic delivery of mRNA or other nucleic acid therapeutics. SNALP compositions include cationic lipids such as MC3 or KC2, comprising a protonatable tertiary amine head group joined to a pair of linear 18 carbon aliphatic chains containing a pair of carbon-carbon double bonds separated by a single methylene group (e.g., linoleic acid). However, while the structure of these hydrocarbon chains, each containing a pair of double bonds separated by a single methylene group, imparts desirable biological properties to the SNALP compositions, this chemical sub-structure also results in the undesired problem of increased sensitivity of the compound to oxidative degradation. For example, FIG.1 is a depiction of the oxidative degradation mechanisms of lipid esters of linoleic acid containing conjugated multiple unsaturations that are particularly sensitive to oxidation. What is needed are novel cationic lipids suitable for use in a SNALP composition, but having enhanced resistance to oxidative degradation. Disclosed herein are compounds, compositions and methods related to the treatment of bacterial infections. As used herein, the term “compound”, “drug” and “active agent” are used interchangeably. Some aspects of the disclosure relate to novel ionizable lipids or bioreducible ionizable lipids. These lipids are cationic (i.e. positively charged) at acidic pH, such as encountered intracellularly following endocytosis or phagocytosis by a cell. The same lipids, and compositions containing them, are near neutral in charge when present at pH 7.4. These lipids may also have a single olefin group present in their alkyl or acyl groups. Some aspects of the disclosure relate to the process for the synthesis of the novel ionizable lipids. 19 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 Other aspects relate to compositions comprising lipidic nanoparticles comprising ionizable cationic lipid, the lipidic nanoparticles containing nucleic acids. In some embodiments, nucleic acids are encapsulated into the lipidic nanoparticles. Aspects of the disclosure provide for improved compositions of ionizable lipid nanoparticles for the delivery of therapeutic nucleic acids to cells. Anionic phospholipids, including phosphatidylserine and phosphatidylglycerol are included in the lipid nanoparticles to increase the transfection efficiency in dendritic cells. The further incorporation of ionizable lipids in an LNP formulation with gem di-substitution of mono-unsaturated alkyl chains (single olefin) on 2-position of 1,3-dioxolane or ketal demonstrated high levels of transfection in human dendritic cells, compared to other ionizable lipids in the same family, and demonstrated good stability to oxidative damage. Definitions For convenience, certain terms employed in the specification, examples, and appended claims are collected here. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. As used herein, the following terms and phrases are intended to have the following meanings: The articles “a” and “an” are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element. As used herein the term "comprising" or "comprises" is used in reference to compositions, methods, and respective component(s) thereof, that are present in a given embodiment, yet open to the inclusion of unspecified elements. As used herein the term "consisting essentially of" refers to those elements required for a given embodiment. The term permits the presence of additional elements that do not materially affect the basic and novel or functional characteristic(s) of that embodiment of the disclosure. The term "consisting of" refers to compositions, methods, and respective components thereof as described herein, which are exclusive of any element not recited in that description of the embodiment. 20 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 The term “comprising” when used in the specification includes “consisting of” and "consisting essentially of". If it is referred to “as mentioned above” or “mentioned above”, “supra” within the description it is referred to any of the disclosures made within the specification in any of the preceding pages. If it is referred to “as mentioned herein”, “described herein”, “provided herein,” or “as mentioned in the present text,” or “stated herein” within the description it is referred to any of the disclosures made within the specification in any of the preceding or subsequent pages. As used herein, the term “about” means acceptable variations within 20%, within 10% and within 5% of the stated value. In certain embodiments, "about" can mean a variation of + / -1%, 2%, 3%, 4%, 5%, 10% or 20%. The term "effective amount" as used herein with respect to a compound or the composition means the amount of active compound (also referred herein as active agent or drug) sufficient to cause a bactericidal or bacteriostatic effect. In some embodiments, the effective amount is a "therapeutically effective amount" meaning the amount of active compound that is sufficient alleviate the symptoms of the bacterial infection being treated. The term "subject" (or, alternatively, "patient") as used herein refers to an animal, preferably a mammal, most preferably a human that receives either prophylactic or therapeutic treatment. The term “administration” or “administering” as used herein includes all means of introducing the compounds or the pharmaceutical compositions to the subject in need thereof, including but not limited to, oral, intravenous, intramuscular, intraperitoneal, subcutaneous, transdermal, inhalation, buccal, ocular, sublingual, vaginal, rectal and the like. Administration of the compound or the composition is suitably parenteral. For example, the compounds or the composition can be preferentially administered intravenously, but can also be administered intraperitoneally or via inhalation like is currently used in the clinic for liposomal amikacin in the treatment of mycobacterium avium (see Shirley et al., Amikacin Liposome Inhalation Suspension: A Review in Mycobacterium avium Complex Lung Disease. Drugs.2019 Apr; 79(5):555-562) The terms “treat,” “treating,” and “treatment,” as used herein, refer to therapeutic or preventative measures such as those described herein. 21 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 The term “pharmaceutically acceptable salt" refers to a relatively non-toxic, inorganic or organic acid addition salt of a compound of the present disclosure which salt possesses the desired pharmacological activity. The term "alkyl" means saturated carbon chains having from one to twenty carbon atoms which may be linear or branched or combinations thereof, unless the carbon chain is defined otherwise. Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, sec- and tert- butyl, pentyl, hexyl, heptyl, octyl, and the like. Unless stated otherwise specifically in the specification, an alkyl group is optionally substituted. The term “phosphatidylserine”, with any of it’s acyl chain compositions, refers to the L- isomer of serine in the headgroup unless specified in a particular example. The term “lipid conjugate” refers to a conjugated lipid that inhibits aggregation of lipid particles. Such lipid conjugates include, but are not limited to, polysarcosine (see e.g. WO2021191265A1 which is herein incorporated by reference in its entirety for all purposes), polyamide oligomers (e.g., ATTA-lipid conjugates), PEG-lipid conjugates, such as PEG coupled to dialkyloxypropyls, PEG coupled to diacylglycerols, PEG coupled to cholesterol, PEG coupled to phosphatidylethanolamines, PEG conjugated to ceramides (see, e.g., U.S. Pat. No.5,885,613, the disclosure of which is herein incorporated by reference in its entirety for all purposes), cationic PEG lipids, and mixtures thereof. PEG can be conjugated directly to the lipid or may be linked to the lipid via a linker moiety. Any linker moiety suitable for coupling the PEG to a lipid can be used including, e.g., non-ester containing linker moieties and ester-containing linker moieties. In preferred embodiments, non-ester containing linker moieties are used. The abbreviations for the ionizable cationic lipids may be truncated in the Examples from that used in the Tables, for example, AKG-UO-1 or AKG-KC2-01 may be referred to as UO1 or KC2-01. The abbreviation UT used in various studies refers to untreated samples. The term “lipidic nanoparticle”, or “LNP”, refers to particles having a diameter of from about 5 to 500 nm. In some embodiments, the lipid nanoparticle comprises one or more active agents. In some embodiments, the lipid nanoparticle comprises a nucleic acid. In some embodiments, the nucleic acid is condensed in the interior of the nanoparticle with a cationic lipid, polymer, or polyvalent small molecule and an external lipid coat that interacts with the biological milieu. Due to the repulsive forces between phosphate groups, nucleic acids are naturally stiff 22 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 polymers and prefer elongated configurations. In the cell, to cope with volume constraints DNA can pack itself in the appropriate solution conditions with the help of ions and other molecules. Usually, DNA condensation is defined as the collapse of extended DNA chains into compact, orderly particles containing only one or a few molecules. By binding to phosphate groups, cationic lipidic can condense DNA by neutralizing the phosphate charges and allow close packing. In some embodiments, the active agent is encapsulated into the LNP. In some embodiments, the active agent can be an anionic compounds, for example, but not limited to DNA, RNA, natural and synthetic oligonucleotides (including antisense oligonucleotides, interfering RNA and small interfering RNA), nucleoprotein, peptide, nucleic acid, ribozyme, DNA- containing nucleoprotein, such as an intact or partially deproteinated viral particles (virions), oligomeric and polymeric anionic compounds other than DNA (for example, acid polysaccharides and glycoproteins)). In some embodiments, the active agent can be intermixed with an adjuvant. In a LNP vaccine product, the active agent is generally contained in the interior of the LNP. In some embodiments, the active agent comprises a nucleic acid. Typically, water soluble nucleic acids are condensed with cationic lipids or polycationic polymers in the interior of the particle and the surface of the particle is enriched in neutral lipids or PEG-lipid derivatives. Additional ionizable cationic lipid may also be at the surface and respond to acidification in the environment by becoming positively charged, facilitating endosomal escape. Ionizable lipids can have different properties or functions with respect to LNPs. Due to the pKa of the amino group, the lipid molecules can become positively charged in acidic conditions. Under these conditions, lipid molecules can electrostatically bind to the phosphate groups of the nucleic acid which allows the formation of LNPs and the entrapment of the nucleic acid. In some embodiments, the pKa can be low enough that it renders the LNP substantially neutral in surface charge in biological fluids, such as blood, which are at physiological pH values. High LNP surface charge is associated with toxicity, rapid clearance from the circulation by the fixed and free macrophages, hemolytic toxicities, including immune activation (Filion et al Biochim Biophys Acta.1997 Oct 23;1329(2):345-56). In some embodiments, pKa can be high enough that the ionizable cationic lipid can adopt a positively charged form at acidic endosomal pH values. This way, the cationic lipids can combine with endogenous endosomal anionic lipids to promote membrane lytic nonbilayer structures such as the hexagonal HII phase, resulting in more efficient intracellular delivery. In some 23 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 embodiments, the pKa ranges between 6.2-6.5. For example, the pKa can be about 6.2, about 6.3, about 6.4, about 6.5. Unsaturated tails also contribute to the lipids’ ability to adopt nonbilayer structures. (Jayaraman et al., Angew Chem Int Ed Engl.2012 Aug 20;51(34):8529-33). Release of nucleic acids from LNP formulations, among other characteristics such as liposomal clearance and circulation half-life, can be modified by the presence of polyethylene glycol and / or sterols (e.g. cholesterol) or other potential additives in the LNP, as well as the overall chemical structure, including pKa of any ionizable cationic lipid included as part of the formulation. The term “bioreducible” refers to compounds that undergo accelerated degradation due to the cleavage of disulfide linkages in a reductive environment. Unlike other nucleic acid therapeutics such as siRNA, the success of mRNA-based therapies depends on the availability of a safe and efficient delivery vehicle that encapsulates the mRNA. mRNA is fragile and needs a protective coating for it to remain active until it reaches its target site. mRNA containing LNPs are a promising vaccine option for Covid-19 immunity (Jackson et al., Preliminary Report. N Engl J Med. 2020 Nov 12;383(20):1920-1931). The efficiency and tolerability of LNPs has been attributed to the amino lipid and unlike many biomaterial applications that may have a required service lifetime of weeks or months, functional LNP mediated delivery of mRNA occurs within hours obviating the need for persistent lipids. Indeed, in applications where chronic dosing is required this will be especially important. It has been demonstrated that LNPs enter cells via endocytosis and accumulate in endolysosomal compartments. The ionizable cationic lipid (ICL) is able to effectively deliver mRNA to the cytosol after endocytosis while being susceptible to enzymatic hydrolysis in late endosomes / lysosomes by lipases or hydrolysis triggered by the reductive environment of the lysosome allowing complete biodegradation. The extracellular space is a relatively oxidative environment, while the intracellular space is a reductive one, allowing a disulfide linked molecule to remain intact in the extracellular space but be rapidly reduced once internalized (Huang et al., Mol Ther.2005 Mar;11(3):409-17, 2005). Some embodiments, provide bioreducible disulfide linked ICL molecules (see compounds 29-36, Table 2) that are stable in LNP formulation and while in circulation but undergo cleavage in the reductive environment of the lysosome. Such compounds and compositions can facilitate rapid biological destruction of the lipids and can prevent potentially toxic accumulation of ICL lipids (as observed in rats with DLin- MC3-DMA (Sabins et al., Mol Ther.2018 Jun 6;26(6):1509-1519). 24 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 The terms “encapsulation” and “entrapped,” as used herein, refer to the incorporation or association of the mRNA, DNA, siRNA or other nucleic acid pharmaceutical agent in or with a lipidic nanoparticle. As used herein, the term “encapsulated” refers to complete encapsulation or partial encapsulation. A siRNA may be capable of selectively knocking down or down regulating expression of a gene of interest. For example, an siRNA could be selected to silence a gene associated with a particular disease, disorder, or condition upon administration to a subject in need thereof of a nanoparticle composition including the siRNA. A siRNA may comprise a sequence that is complementary to an mRNA sequence that encodes a gene or protein of interest. The term “mol%" with regard to cholesterol refers to the molar amount of cholesterol relative to the sum of the molar amounts of cholesterol and non-PEGylated phospholipid expressed in percentage points. For example, “55 mol.% cholesterol” in a liposome containing cholesterol and HSPC refers to the composition of 55 mol. parts of cholesterol per 45 mol. parts of HSPC. The term “mol%" with regard to PEG-lipid refers to the ratio of the molar amount of PEG- lipid and non-PEGylated phospholipid expressed in percentage points. For example, “5 mol.% PEG-DSPE” in a LNP containing HSPC and PEG-DSPE refers to the composition having 5 mol. parts of PEG-DSPE per 100 mol. parts of HSPC. In some embodiments, “mol%" with regard to cholesterol refers to the molar amount of cholesterol relative to the sum of the molar amounts of total lipid expressed in percentage points. For example, “40.5 mol % cholesterol” in an LNP composition containing cholesterol, an ICL, DSPC, PS, cholesterol, and PEG-DMG refers to the composition of 40.5 mol. parts of cholesterol per 59.5 mol. parts of the ICL, DSPC, PS, and PEG-DMG components combined. In some embodiments, “mol%" with regard to conjugated lipid refers to the ratio of the molar amount of conjugated lipid expressed in percentage points. For example, “1.5 mol % PEG- DMG” in an LNP containing cholesterol, an ICL, DSPC, PS, cholesterol, and PEG-DMG refers to the composition of 1.5 mol. parts of PEG-DMG per 98.5 mol. parts of the combined ICL, DSPC, PS, and cholesterol components. In some embodiments, “mol%" with regard to the PS lipid refers to the ratio of the molar amount of PS lipid expressed in percentage points. For example, “5 mol % DPPS” in an LNP containing cholesterol, an ICL, DSPC, DPPS, cholesterol, and PEG-DMG refers to the composition of 5 mol. parts of PEG-DMG per 95 mol. parts of the combined ICL, DSPC, PEG- DMG, and cholesterol components. 25 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 In some embodiments, “mol%" with regard to ICL refers to the ratio of the molar amount of ICL expressed in percentage points. For example, “48 mol % PEG-DMG” in an LNP containing cholesterol, KC3-OA (e.g. an example of an ICL), DSPC, PS, cholesterol, and PEG-DMG refers to the composition of 48 mol. parts of the ICL per 52 mol. parts of the combined PEG-DMG, DSPC, PS, and cholesterol components. As used herein, the term “pharmaceutically acceptable carrier, diluent or excipient” includes without limitation any adjuvant, carrier, excipient, glidant, sweetening agent, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier which has been approved by the United States Food and Drug Administration as being acceptable for use in humans or domestic animals. Various aspects and embodiments are described in further detail in the following subsections. Ionizable Cationic Lipids Provided herein are compounds useful in the preparation of lipid nanoparticle (LNP) compositions. In an embodiment, the LNP comprises an ionizable cationic lipid. As used herein “ionizable cationic lipid”, “ionizable lipid” and “ICL” are used interchangeably. An ICL is a lipid that comprises an ionizable moiety capable of bearing a charge (e.g., a positive charge e.g., a cationic lipid) under certain conditions (e.g., at a certain pH range, e.g., under physiological conditions). The ionizable moiety may comprise an amine, and preferably a substituted amine. An ionizable lipid may be a cationic lipid or an anionic lipid. In addition to an ionizable moiety, an ionizable lipid may contain an alkyl or alkenyl group, e.g., greater than six carbon atoms in length (e.g., greater than about 8 carbons, 10 carbons, 12 carbons, 14 carbons, 16 carbons, 18 carbons, 20 carbons or more in length). Additional ionizable lipids that may be included in an LNP described herein are disclosed in Jayaraman et al. (Angew. Chem. Int. Ed. 51:8529-8533 (2012)), Semple et al. Nature Biotechnol.28:172-176 (2010)), and U.S. Patent Nos.8,710,200 and 8,754,062, each of which is incorporated herein by reference in its entirety. An LNP may comprise an ionizable lipid at a concentration greater than about 0.1 mol%, e.g., of the total lipid content of the LNP. In an embodiment, the LNP comprises an ionizable lipid at a concentration of greater than about 1 mol%, about 2mol%, about 4mol%, about 8mol%, about 20mol%, about 40mol%, about 50mol%, about 60mol%, about 80mol%, e.g., of the total lipid 26 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 content of the LNP. In an embodiment, the LNP comprises an ionizable lipid at a concentration of greater than about 20mol%, about 40mol%, or about 50mol%. In an embodiment, the LNP comprises an ionizable lipid at a concentration between about 1mol% to about 95mol%, e.g., of the total lipid content of the LNP. In an embodiment, the LNP comprises an ionizable lipid at a concentration between about 2mol% to about 90mol%, about 4mol% to about 80mol%, about 10mol% to about 70mol%, about 20mol% to about 60mol%, about 40mol% to about 55mol%, e.g., of the total lipid content of the LNP. In an embodiment, the LNP comprises an ionizable lipid at a concentration between about 20mol% to about 60mol%. In an embodiment, the LNP comprises an ionizable lipid at a concentration between about 40 mol% to about 55 mol%. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I): , 3 or 4; R2 and R3 are each independently (C1-C4) alkyl optionally substituted with hydroxyl; and n is an integer equal to 2, 3 or 4. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I), wherein the total length of the R1 hydrocarbon chain is C15 - C18. In some embodiments, the total length of the R1 hydrocarbon chain is C16- C18. In some embodiments, the total length of the R1hydrocarbon chain is C16or C18. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I), wherein a is 0 and b is 1, 2, 3 or 4. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I), wherein a is 0 and b is 1 or 3. In some embodiments, 27 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I), wherein a is 0 and b is 1. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I), wherein a is 0 and b is 3. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I), wherein a is 1 and b is 1, 2, 3 or 4. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I), wherein a is 1 and b is 1 or 3. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I), wherein a is 1 and b is 1. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I), wherein a is 1 and b is 3. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I), wherein R10and R12are the same. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I), wherein R10 and R12 are each (C1-C4)alkyl optionally substituted with hydroxyl. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I), wherein R10and R12 are each (C1-C4)alkyl. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I), wherein R10and R12are each methyl. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I), wherein R10 and R12 are each ethyl. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I), wherein R10and R12are each independently selected from methyl or ethyl. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I), wherein R10and R12are each independently selected from methyl, ethyl, -(CH2)(CH2)OH, and - (CH2)2(CH2)OH. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I), wherein a is 0 or 1; b is 1, 2, 3 or 4, provided the sum a+b is 1, 2, 3 or 4; R2and R3are each methyl; and n is an integer equal to 2, 3 28 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 or 4. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I), wherein a is 0 or 1; b is 1, 2, 3 or 4, provided the sum a+b is 1, 2, 3 or 4; R2 and R3 are each methyl; and n is an integer equal to 2 or 3. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I), wherein a is 0 or 1; b is 1, 2, 3 or 4, provided the sum a+b is 1, 2, 3 or 4; R2 and R3 are each methyl; and n is an integer equal to 2. n some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I), wherein a is 0 or 1; b is 1, 2, 3 or 4, provided the sum a+b is 1, 2, 3 or 4; R2 and R3 are each methyl; and n is an integer equal to 3. In some embodiments, an ionizable cationic lipid comprises the chemical structure of Formula (II): , or a pharmaceutically acceptable O R22, 4; R22is a hydrocarbon chain with a single olefin and a total length of C15-C18; and each of R10 and R12 is independently (C1-C4)alkyl optionally substituted with hydroxyl. In some aspects, R22in Formula (II) is a polyene hydrorcarbon chain of Formula A. In some aspects, R10and R12in Formula (II) are each independently selected from methyl, ethyl, propyl, -(CH2)(CH2)OH, and -(CH2)2(CH2)OH. In some aspects, R10and R12are each independently methyl in Formula (II). In some aspects, R10 and R12 are each independently ethyl in Formula (II). In some aspects, at least one of R10 and R12 is n-propyl optionally substituted with hydroxyl in Formula (II). In some aspects, R10is methyl and R12is selected from methyl, ethyl, - (CH2)(CH2)OH, and -(CH2)2(CH2)OH in Formula (II). In some aspects, R10 is methyl and R12 is selected from -(CH2)(CH2)OH, and -(CH2)2(CH2)OH in Formula (II). In some aspects, R10 is methyl and R12is selected from -(CH2)(CH2)OH, and -(CH2)2(CH2)OH in a compound comprising 29 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 the chemical structure of Formula (II). In some aspects, R10and R12are independently selected from methyl or ethyl, optionally substituted with one or more hydroxyl in Formula (II). In some aspects, one or both of R10 and R12 in Formula (II) are -(CH2)(CH2)OH, or -(CH2)2(CH2)OH in Formula (II). In some aspects, R10is methyl and R12is methyl or ethyl substituted with hydroxyl in Formula (II). In some aspects, one or both of R10in Formula (II) is methyl and R12is - (CH2)(CH2)OH in Formula (II). In some aspects, one or both of R10 in Formula (II) is methyl and R12is -(CH2)2(CH2)OH in Formula (II). In some embodiments, the compounds have the structure of the compounds listed in the tables below. Table 1A and Table 1B show examples of cationic lipids. Table 2 shows examples of bioreducible cationic lipids. Table 1A. Exemplary cationic lipids 30 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 31 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 Table 1A. Exemplary cationic lipids (continued) 32 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 Table 1A. Exemplary cationic lipids (continued) 33 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 Table 1A. Exemplary cationic lipids (continued) 34 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 Table 1B Additional Exemplary cationic lipids 35 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 Table 2. Exemplary bioreducible cationic lipids 36 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 Table 2. Exemplary bioreducible cationic lipids (continued) In some embodiments, the ionizable lipid encapsulate the nucleic acid. In some embodiments, the ionizable lipid encapsulate the nucleic acid in a LNP formulation. In some embodiments, the nucleic acid is a siRNA molecule. In some embodiments, the nucleic acid is a mRNA molecule. In some embodiments, the nucleic acid is a DNA molecule. In some embodiments, compositions further comprising ligands, such as antibody conjugates, directed against cell surface receptors to target lipid nanoparticles in a highly specific manner to dendritic cells are provided. In some embodiments, the composition further comprises a targeting ligand, wherein the targeting ligand is oriented to the outside of the nanoparticle. In some embodiments, the targeting ligand is an antibody. In some embodiments, the lipidic nanoparticles are in an aqueous medium. In some embodiments, the nucleic acid is entrapped in the lipidic nanoparticle with a compound disclosed herein, including compounds of Formula I, II, III, IV-B, V-A-1 or combinations thereof, wherein the nucleic acid is either RNA or DNA. In some embodiments, the nucleic acid is entrapped in the lipidic nanoparticle with a compound disclosed herein, including 37 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 compounds of disclosed herein or combinations thereof, wherein the nucleic acid is either RNA or DNA. In some embodiments, the nucleic acid is mRNA. In some embodiments, the nucleic acid is siRNA. In some embodiments, the nucleic acid is DNA. In some embodiments, the lipidic nanoparticle comprises a membrane comprising phosphatidylcholine and a sterol. In some embodiments, the sterol is cholesterol. In other embodiments, the sterol is beta-sitosterol. In some embodiments, the lipidic nanoparticle comprises a membrane comprising phosphatidylcholine, ionizable cationic lipid (ICL). In some embodiments, the ICL have a structure of Formula I, II, III, IV-B, V-A-1, and cholesterol, wherein the membrane separates the inside of the lipidic nanoparticles from the aqueous medium. In some embodiment, the ICL have a structure as shown in Table 1A and Table 2. In some embodiment, the ICL have a structure as shown in Table 1B. In some embodiments, the phosphatidylcholine is distearoylphosphatidylcholine (DSPC) or hydrogenated soy phosphatidylcholine (HSPC). In some embodiments, the ionizable cationic lipid to cholesterol molar ratios is from about 65:35 to 40:60. In some embodiments, the ICL to cholesterol molar ratio is from about 60:40 to about 45:55. In some embodiments, the phosphatidylcholine to cholesterol molar ratio is from about 1:5 to about 1:2. In some embodiments, the membrane further comprises a polymer-conjugated lipid. In some embodiments, the lipidic nanoparticle comprises ICL, DSPC, cholesterol and polymer-conjugated lipid in a about 49.5:10.3:39.6:2.5 molar ratio. In some embodiments, the polymer-conjugated lipid is PEG(2000)-dimyristoylglycerol (PEG-DMG) or PEG(Mol. weight 2,000)-dimyristoylphosphatidylethanolamine (PEG-DMPE). In some embodiments the percentage of oxidative degradation products for the ionizable lipid is less than 50 % of that for a DLin-KC2-DMA or DLin-MC3-DMA control formulation. In some embodiments, the composition is a liquid pharmaceutical formulation for parenteral administration. In some embodiments, the composition is a liquid pharmaceutical formulation for subcutaneous, intramuscular, or intradermal administration. In some embodiments, the composition is in the form of a lyophilized powder, that is subsequently reconstituted with aqueous medium prior to administration. Other aspects of the disclosure relate to a method of preventing a bacterial or viral infection, 38 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 the method comprising administering to a subject in need thereof an effective amount of the composition provided herein to elicit an immune response. Some embodiments provide methods of vaccinating a subject in need thereof, the method comprising administering the composition comprising a nucleic acid encoding an antigenic protein. In some embodiments, the composition is administered subcutaneously, intramuscularly, or intradermally. In some embodiments, the bacterial infection is Mycobacterium tuberculosis infection. In some embodiments, the bacterial infection is a form of nontuberculosis mycobacterium. In some embodiments, the viral infection is a coronavirus. In some embodiments, the coronavirus is SARS-CoV, MERS-CoV or SARS-CoV-2 In some embodiments, the viral infection is HIV / AIDs. In some embodiments, the lipidic nanoparticle is administered parenterally. In some embodiments, the lipidic nanoparticle composition is administered as part of a single injection. The present disclosure features a lipid nanoparticle comprising nucleic acids such as DNA, mRNA, siRNA, antisense oligonucleotides, CRISPR components such as a guide RNA (gRNA or sgRNA) and a CRISPR-associated endonuclease (Cas protein) and a lipid. Exemplary lipids include ionizable cationic lipids (ICLs), phospholipids, sterol lipids, alkylene glycol lipids (e.g., polyethylene glycol lipids), sphingolipids, glycerolipids, glycerophospholipids, prenol lipids, saccharolipids, fatty acids, and polyketides. In some embodiments, the LNP comprises a single type of lipid. In some embodiments, the LNP comprises a plurality (e.g. two or more) of lipids. An LNP may comprise one or more of an ionizable cationic lipid, a phospholipid, a sterol, or an alkylene glycol lipid (e.g., a polyethylene glycol lipid). In some embodiments, an LNP further comprises an ionizable lipid having a structure of Formula (IV-A), or a pharmaceutically acceptable salt thereof, ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 wherein each of R10 and with hydroxyl; O is In some embodiments, v equals 0 for compounds of Formula (III). In some embodiments, v equals 1 for compounds of Formula (III). In some embodiments, v equals 1 and q1 equals 1 for compounds of Formula (III). In some embodiments, v equals 1 and q1 equals 2 for compounds of Formula (III). In some embodiments, the sum of a and c is 6, 7, 8 or 9 in R22for compounds of Formula (III). In some embodiments, the sum of a and c is 6 in R22for compounds of Formula (III). In some embodiments, the sum of a and c is 7 in R22for compounds of Formula (III). In some embodiments, the sum of a and c is 9 in R22for compounds of Formula (III). In some embodiments, v equals 0 and the sum of a and c is 6, 7, 8 or 9 in R22for compounds of Formula (III). In some embodiments, v equals 0 and the sum of a and c is 6 in R22for compounds of Formula (IV-B). In some embodiments, v equals 0 and the sum of a and c is 7 in R22for compounds of Formula (III). In some embodiments, v equals 0 and the sum of a and c is 9 in R22for compounds of Formula (III). 40 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 In some embodiments, R10and R12are independently selected from methyl, ethyl, - (CH2)(CH2)OH, and -(CH2)2(CH2)OH for compounds of Formula (III). In some embodiments, R10 and R12 are each methyl and the sum of a and c is 6, 7, 8 or 9 in R22for compounds of Formula (III). In some embodiments, R10and R12are each methyl, v is 0 and the sum of a and c is 6, 7, 8 or 9 in R22for compounds of Formula (III). In some embodiments, v equals 0 and R22compounds of Formula (IV-B). In some embodiments, v equals 0 and R22, and the sum of a and c is 7 or 9 for compounds of Formula (III). In some 0 and R22is v some embodiments, v equals 0 and R22compounds of Formula (III). In an embodiment, the LNP comprises a phospholipid. A phospholipid is a lipid that comprises a phosphate group and at least one alkyl, alkenyl, or heteroalkyl chain. A phospholipid may be naturally occurring or non-naturally occurring (e.g., a synthetic phospholipid). A 41 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 phospholipid may comprise an amine, amide, ester, carboxyl, choline, hydroxyl, acetal, ether, carbohydrate, sterol, or a glycerol. In some embodiments, a phospholipid may comprise a phosphocholine, phosphosphingolipid, or a plasmalogen. Exemplary phospholipids include 1,2- dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-distearoyl-sn-glycero-3- phosphocholine (DSPC), hydrogenated soy phosphatidylcholine (HSPC), 1,2-dilauroyl-sn- glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2- distearoyl-sn-glycero-3-phosphoethanolamine (DSPE), 1-myristoyl-2-oleoyl-sn-glycero-3- phosphocholine (MOPC), 1,2-diarachidonoyl-sn-glycero-3-phosphocholine (DAPC), 1- palmitoyl-2-linoleoyl-sn-glycero-3-phosphatidylcholine (PLPC), 1-palmitoyl-2-oleoyl-glycero-3- phosphocholine (POPC), 1-stearoyl-2-myristoyl-sn-glycero-3-phosphocholine (SMPC), 1- palmitoyl-2-myristoyl-sn-glycero-3-phosphocholine (PMPC), bis(monoacylglycerol)phosphate (BMP), L-α-phosphatidylcholine, 1,2-Diheptadecanoyl-sn-glycero-3-phosphorylcholine (DHDPC), and 1-stearoyl-2-arachidonoyl-sn-glycero-3-phosphocholine (SAPC). Additional phospholipids that may be included in an LNP described herein are disclosed in Li, J. et al. (Asian J. Pharm. Sci.10:81-98 (2015)), which is incorporated herein by reference in its entirety. In some embodiments, the phospholipid is 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC). In some embodiments, the phospholipid is 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC). In some embodiments, the phospholipid is 1,2-dipalmitoyl-sn-glycero-3- phosphocholine (DPPC). In some embodiments, the phospholipid is 1,2-dioleoyl-sn-glycero-3- phosphoethanolamine (DOPE). Incorporation of phosphatidylserine The LNP (e.g., as described herein) may comprise one or more of the following components: (i) Ionizable cationic lipid (ICL) containing a C16 alkyl or C16 alkenyl group or C18 alkyl or C18 alkenyl group at a concentration between about 1mol% to about 95mol% (or any value therebetween, e.g. about 20mol% to about 80mol%); (ii) A phospholipid at a concentration between 0.1mol% to about 20 mol% (or any value there between, e.g. between about 2.5 mol% to about 10 mol%) where the phospholipid also contains C16 or C18 alkyl or alkenyl groups; (iii) cholesterol at a concentration between about 1mol% to about 95mol% (or any value therebetween, e.g. about 20mol% to about 80mol%); (iv) a phosphatidylserine (PS) or phosphatidylglycerol (PG) added to the LNP lipid formulation at a concentration between about 0.5 mol% to about 20 mol%, 42 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 about 2.5 mol% to about 10 mol%, about 4 mol% to about 8 mol%, or any value therebetween of the total lipid content of the LNP, and (v) a polyethyleneglycol (PEG)-2000-containing lipid (e.g., DPG-PEG2000, DPPE-PEG2000, DMPE-PEG2000, DMG-PEG2000) at a concentration between about 0.1mol% to about 5 mol% (or any value therebetween, e.g. between about 1 mol% to about 2.5 mol%). In some embodiments, the LNP comprises two of (i)-(v). In some embodiments, the LNP comprises three of (i)-(v). In some embodiments, the LNP comprises four of (i)-(v). In some embodiments, the LNP comprises each of (i)-(v). In some embodiments, the LNP comprises (i) and (ii). In some embodiments, the LNP comprises (i) and (iii). In some embodiments, the LNP comprises (i) and (v). In some embodiments, the LNP comprises (ii) and (iii). In some embodiments, the LNP comprises (ii) and (v). In some embodiments, the LNP comprises (iii) and (iv). In some embodiments, the LNP comprises (iii) and (v). In some embodiments, the LNP comprises (i), (ii), and (iii). In some embodiments, the LNP comprises (i), (ii), and (v). In some embodiments, the LNP comprises (ii), (iii), and (v). In some embodiments, the LNP comprises (ii), (iii), (iv) and (v). In an embodiment, the LNP consists or consists essentially of four of (i)-(v). In an embodiment, the LNP consists or consists essentially of each of (i)-(v). In some embodiments, the LNP consists or consists essentially of (i) and (ii). In some embodiments, the LNP consists or consists essentially of (i) and (iii). In some embodiments, the LNP consists or consists essentially of (i) and (v). In some embodiments, the LNP consists or consists essentially of (ii) and (iii). In some embodiments, the LNP comprises (ii) and (v). In some embodiments, the LNP consists or consists essentially of (iii) and (iv). In some embodiments, the LNP consists or consists essentially of (iii) and (v). In some embodiments, the LNP consists or consists essentially of (i), (ii), and (iii). In some embodiments, the LNP consists or consists essentially of (i), (ii), and (v). In some embodiments, the LNP comprises (ii), (iii), and (v). In some embodiments, the LNP consists or consists essentially of (ii), (iii), (iv) and (v). An LNP may comprise a phospholipid at a concentration greater than about 0.1mol%, e.g., of the total lipid content of the LNP. In an embodiment, the LNP comprises a phospholipid at a concentration of greater than about 0.5mol%, about 1mol%, about 1.5mol%, about 2mol%, about 3mol%, about 4mol%, about 5mol%, about 6mol%, about 8mol%, about 10mol%, about 12mol%, about 15mol%, about 20mol%, about 50mol%, e.g., of the total lipid content of the LNP. In an embodiment, the LNP comprises a phospholipid at a concentration of greater than about 1mol%, about 5mol%, or about 10mol%. In an embodiment, the LNP comprises a phospholipid at a 43 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 concentration between about 0.1mol% to about 50mol%, e.g., of the total lipid content of the LNP. In an embodiment, the LNP comprises a phospholipid at a concentration between about 0.5mol% to about 40mol%, about 1mol% to about 30mol%, about 5mol% to about 25mol%, about 10mol% to about 20mol%, about 10mol% to about 15mol%, or about 15mol% to about 20mol%, e.g., of the total lipid content of the LNP. In an embodiment, the LNP comprises a phospholipid at a concentration between about 5mol% to about 25mol%. In an embodiment, the LNP comprises a phospholipid at a concentration between about 10mol% to 20mol%. In an embodiment, the LNP comprises a sterol or ionizable sterol molecule. A sterol is a lipid that comprises a polycyclic structure and an optionally a hydroxyl or ether substituent, and may be naturally occurring or non-naturally occurring (e.g., a synthetic sterol). Sterols may comprise no double bonds, a single double bond, or multiple double bonds. Sterols may further comprise an alkyl, alkenyl, halo, ester, ketone, hydroxyl, amine, polyether, carbohydrate, or cyclic moiety. An exemplary listing of sterols includes cholesterol, dehydroergosterol, ergosterol, campesterol, β-sitosterol, stigmasterol, lanosterol, dihydrolanosterol, desmosterol, brassicasterol, lathosterol, zymosterol, 7-dehydrodesmosterol, avenasterol, campestanol, lupeol, and cycloartenol. In some embodiments, the sterol comprises cholesterol, dehydroergosterol, ergosterol, campesterol, β-sitosterol, or stigmasterol. Additional sterols that may be included in an LNP described herein are disclosed in Fahy, E. et al. (J. Lipid. Res.46:839-862 (2005). Ionizable sterols In some embodiments, an LNP comprises a sterol. In some embodiments, the sterol is cholesterol. In some embodiments, the sterol is dehydroergosterol. In some embodiments, the sterol is ergosterol. In some embodiments, the sterol is campesterol. In some embodiments, the sterol is β-sitosterol. In some embodiments, the sterol is stigmasterol. In some embodiments, the sterol is a corticosteroid. (e.g., corticosterone, hydrocortisone, cortisone, or aldosterone). In some embodiments, the ionizable lipid can be a branched ionizable lipid selected from ALC-0315 and SM-102: 44 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 . An LNP may comprise a sterol at a concentration greater than about 0.1mol%, e.g., of the total lipid content of the LNP. In an embodiment, the LNP comprises a sterol at a concentration greater than about 0.5mol%, about 1mol%, about 5mol%, about 10mol%, about 15mol%, about 20mol%, about 25mol%, about 35mol%, about 40mol%, about 45mol%, about 50mol%, about 55mol%, about 60mol%, about 65mol%, or about 70mol%, e.g., of the total lipid content of the LNP. In an embodiment, the LNP comprises a sterol at a concentration greater than about 10mol%, about 15mol%, about 20mol%, or about 25mol%. In an embodiment, the LNP comprises a sterol at a concentration between about 1mol% to about 95mol%, e.g., of the total lipid content of the LNP. In an embodiment, the LNP comprises a sterol at a concentration between about 5mol% to about 90mol%, about 10mol% to about 85mol%, about 20mol% to about 80mol%, about 20mol% to about 60mol%, about 20mol% to about 50mol%, or about 20mol% to 40mol%, e.g., of the total lipid content of the LNP. In an embodiment, the LNP comprises a sterol at a concentration between about 20mol% to about 50mol%. In an embodiment, the LNP comprises a sterol at a concentration between about 30mol% to about 60mol%. In some embodiments, the sterol is cholesterol. In other embodiments, the sterol is beta- sitosterol. In some embodiments, the LNP comprises an alkylene glycol-containing lipid. An alkylene glycol-containing lipid is a lipid that comprises at least one alkylene glycol moiety, for 45 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 example, a methylene glycol or an ethylene glycol moiety. In some embodiments, the alkylene glycol-containing lipid comprises a polyethylene glycol (PEG). An alkylene glycol-containing lipid may be a PEG-containing lipid. Polymer-conjugated lipids may include poly(ethylene glycol)-conjugated (pegylated)phospholipids (PEG-lipids) such as PEG(Mol. weight 2,000) methoxy-poly(ethylene glycol)-1,2-distearoyl-sn-glycerol (PEG-DSG), PEG(Mol. weight 2,000) methoxy-poly(ethylene glycol)-1,2-palmitoyl-sn-glycerol (PEG-DPG), PEG(Mol. weight 2,000) 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (PEG-DSPE) or N-palmitoyl-sphingosine-1-{succinyl[methoxy(polyethylene glycol)2000]} (PEG-ceramide). The molecular weight of the PEG portion in the PEG-lipid component can also vary from 500-10,000 g / mol, from 1,500-6000 g / mol, but is preferably about 2,000 MW. Other polymers used for conjugation to lipid anchors may include poly(2-methyl-2-oxazoline) (PMOZ), poly(2-ethyl-2-oxazoline) (PEOZ), poly-N-vinylpyrrolidone (PVP), polyglycerol, poly(hydroxyethyl L-asparagine) (PHEA), and poly(hydroxyethyl L-glutamine) (PHEG). A PEG-containing lipid may further comprise an amine, amide, ester, carboxyl, phosphate, choline, hydroxyl, acetal, ether, heterocycle, or carbohydrate. PEG-containing lipids may comprise at least one alkyl or alkenyl group, e.g., greater than six carbon atoms in length (e.g., greater than about 8 carbons, 10 carbons, 12 carbons, 14 carbons, 16 carbons, 18 carbons, 20 carbons or more in length), e.g., in addition to a PEG moiety. In an embodiment, a PEG-containing lipid comprises a PEG moiety comprising at least 20 PEG monomers, e.g., at least 30 PEG monomers, 40 PEG monomers, 45 PEG monomers, 50 PEG monomers, 100 PEG monomers, 200 PEG monomers, 300 PEG monomers, 500 PEG monomers, 1000 PEG monomers, or 2000 PEG monomers. Exemplary PEG-containing lipids include PEG-DMG (e.g., DMG-PEG2k), PEG-c- DMG, PEG-DSG, PEG-DPG, PEG-DSPE, PEG-DMPE, PEG-DPPE, PEG-DOPE, and PEG- DLPE. In some embodiments, the PEG-lipids include PEG-DMG (e.g., DMG-PEG2k), PEG-c- DMG, PEG-DSG, and PEG-DPG. Additional PEG-lipids that may be included in an LNP described herein are disclosed in Fahy, E. et al. (J. Lipid. Res. 46:839-862 (2005) which is incorporated herein by reference in its entirety. In some embodiments, PEG-containing conjugated lipid is PEG(2000)- dimyristoylglycerol (PEG-DMG), 1,2-dilauroyl-sn-glycero-3-phosphoethanolamine-N- [methoxy(polyethylene glycol)-2000] (PEG-DLPE), or PEG(2000)-dilauroylglycerol (PEG- DLG). 46 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 In some embodiments, the PEG-lipid is PEG-DMG (e.g., DMG-PEG2k). In some embodiments, the PEG-lipid is α-(3’-{[1,2-di(myristyloxy)propanoxy] carbonylamino}propyl)-ω- methoxy, polyoxyethylene (PEG-c-DMG). In some embodiments, the PEG-lipid is PEG-DSG. In some embodiments, the PEG-lipid is PEG-DPG. In some embodiments, the PEG-lipid is mPEG- 1,2-dilauroylglycerol (PEG-DLG). An LNP may comprise an alkylene glycol-containing lipid at a concentration greater than about 0.1mol%, e.g., of the total lipid content of the LNP. In an embodiment, the LNP comprises an alkylene glycol-containing lipid at a concentration of greater than about 0.5mol%, about 1mol%, about 1.5mol%, about 2mol%, about 3mol%, about 4mol%, about 5mol%, about 6mol%, about 8mol%, about 10mol%, about 12mol%, about 15mol%, about 20mol%, about 50mol%, e.g., of the total lipid content of the LNP. In an embodiment, the LNP comprises an alkylene glycol- containing lipid at a concentration of greater than about 1mol%, about 4mol%, or about 6mol%. In an embodiment, the LNP comprises an alkylene glycol-containing lipid at a concentration between about 0.1mol% to about 50mol%, e.g., of the total lipid content of the LNP. In an embodiment, the LNP comprises an alkylene glycol-containing lipid at a concentration between about 0.5mol% to about 40mol%, about 1mol% to about 35mol%, about 1.5mol% to about 30mol%, about 2mol% to about 25mol%, about 2.5mol% to about 20%, about 3mol% to about 15mol%, about 3.5mol% to about 10mol%, or about 4mol% to 9mol%, e.g., of the total lipid content of the LNP. In an embodiment, the LNP comprises an alkylene glycol-containing lipid at a concentration between about 1.5mol% to about 4mol%. In an embodiment, the LNP comprises an alkylene glycol-containing lipid at a concentration between about 3.5mol% to about 10mol%. In an embodiment, the LNP comprises an alkylene glycol-containing lipid at a concentration between about 4mol% to 9mol%. In some embodiments, the LNP comprises at least two types of lipids. In an embodiment, the LNP comprises two of an ionizable lipid, a phospholipid, a sterol, and an alkylene glycol- containing lipid. In some embodiments, the LNP comprises at least three types of lipids. In an embodiment, the LNP comprises three of an ionizable lipid, a phospholipid, a sterol, and an alkylene glycol-containing lipid. In some embodiments, the LNP comprises at least four types of lipids. In an embodiment, the LNP comprises each of an ionizable lipid, a phospholipid, a sterol, and an alkylene glycol-containing lipid. 47 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 The LNP (e.g., as described herein) may comprise one or more of the following components: (i) an ionizable cationic lipid at a concentration between about 1mol% to about 95mol% (e.g. about 20mol% to about 80mol%); (ii) a phospholipid at a concentration between 0.1mol% to about 50mol% (e.g. between about 2.5mol% to about 20mol%); (iii) a sterol at a concentration between about 1mol% to about 95mol% (e.g. about 20mol% to about 80mol%); and (iv) a PEG-containing lipid at a concentration between about 0.1mol% to about 50mol% (e.g. between about 2.5mol% to about 20mol%). In some embodiments, the LNP comprises one of (i)- (iv). In some embodiments, the LNP comprises two of (i)-(iv). In some embodiments, the LNP comprises three of (i)-(iv). In some embodiments, the LNP comprises each of (i)-(iv). In some embodiments, the LNP comprises (i) and (ii). In some embodiments, the LNP comprises (i) and (iii). In some embodiments, the LNP comprises (i) and (iv). In some embodiments, the LNP comprises (ii) and (iii). In some embodiments, the LNP comprises (ii) and (iv). In some embodiments, the LNP comprises (iii) and (iv). In some embodiments, the LNP comprises (i), (ii), and (iii). In some embodiments, the LNP comprises (i), (ii), and (iv). In some embodiments, the LNP comprises (ii), (iii), and (iv). The LNP (e.g., as described herein) may comprise one or more of the following components: (i) Ionizable cationic lipid (ICL) at a concentration between about 1mol% to about 95mol% (e.g. about 20mol% to about 80mol%); (ii) DSPC at a concentration between 0.1mol% to about 50mol% (e.g. between about 2.5mol% to about 20mol%); (iii) cholesterol at a concentration between about 1mol% to about 95mol% (e.g. about 20mol% to about 80mol%); and (iv) DMG-PEG2k at a concentration between about 0.1mol% to about 50mol% (e.g. between about 2.5mol% to about 20mol%). In some embodiments, the LNP comprises two of (i)-(iv). In some embodiments, the LNP comprises three of (i)-(iv). In some embodiments, the LNP comprises each of (i)-(iv). In some embodiments, the LNP comprises (i) and (ii). In some embodiments, the LNP comprises (i) and (iii). In some embodiments, the LNP comprises (i) and (iv). In some embodiments, the LNP comprises (ii) and (iii). In some embodiments, the LNP comprises (ii) and (iv). In some embodiments, the LNP comprises (iii) and (iv). In some embodiments, the LNP comprises (iii) and (iv). In some embodiments, the LNP comprises (i), (ii), and (iii). In some embodiments, the LNP comprises (i), (ii), and (iv). In some embodiments, the LNP comprises (ii), (iii), and (iv). 48 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 In an embodiment, the LNP comprises a ratio of ionizable lipid to phospholipid of about 50:1 to about 1:1 (e.g., 40:1, 32:3, 6:1, 7:1, 5:1, 24:5, 26:5, 10:3, 15:2, 16:7, 18:1, 3:1, 3:2, or 1:1). In an embodiment, the LNP comprises a ratio of ionizable lipid to phospholipid of about 15:2. In an embodiment, the LNP comprises a ratio of ionizable lipid to phospholipid of about 5:1. In an embodiment, the LNP comprises a ratio of ionizable lipid to a sterol of about 10:1 to about 1:10 (e.g., 9:1, 8:1, 8:7, 7:1, 7:5, 7:3, 6:1, 6:5, 5:1, 5:3, 4:1, 4:3, 3:1, 2:1, 1:1, 1:2, 1:3, 3:4, 1:4, 3:5, 1:5, 4:5, 1:6, 5:6, 7:6, 7:8, or 8:9). In an embodiment, the LNP comprises a ratio of ionizable lipid to an alkylene-containing lipid of about 1:10 to about 10:1 (e.g., 1:9, 1:8, 7:8, 7:1, 7:5, 7:3, 6:1, 6:5, 5:1, 5:3, 4:1, 4:3, 3:1, 2:1, 1:1, 1:2, 1:3, 3:4, 1:4, 3:5, 1:5, 4:5, 1:6, 5:6, 7:6, 7:8, or 8:9). In an embodiment, the LNP comprises a ratio of phospholipid to an alkylene-containing lipid of about 10:1 to about 1:10 (e.g., 9:1, 8:1, 8:7, 7:1, 7:5, 7:3, 6:1, 6:5, 5:1, 5:3, 4:1, 4:3, 3:1, 2:1, 1:1, 1:2, 1:3, 3:4, 1:4, 3:5, 1:5, 4:5, 1:6, 5:6, 7:6, 7:8, or 8:9). In an embodiment, the LNP comprises a ratio of a sterol to an alkylene-containing lipid of about 50:1 to about 1:1 (e.g., 40:1, 32:3, 6:1, 7:1, 5:1, 24:1, 22:1, 20:1, 22:5, 24:5, 26:5, 10:3, 15:2, 16:7, 18:1, 3:1, 3:2, or 1:1). In some embodiments, a LNP (e.g., described herein) comprises two of an ionizable lipid, a phospholipid, a sterol, and an alkylene glycol-containing lipid (e.g., PEG-containing lipid). In another embodiment, a LNP (e.g., described herein) comprises three of an ionizable lipid, a phospholipid, a sterol, and an alkylene glycol-containing lipid (e.g., PEG-containing lipid). In some embodiments, LNP (e.g., described herein) comprises each of an ionizable lipid, a phospholipid, a sterol, and an alkylene glycol-containing lipid (e.g., PEG-containing lipid). In some embodiments, an LNP described herein has a diameter between 5 and 500 nm, e.g., between 10 and 400 nm, 20 and 350 nm, 25 and 325 nm, 30 and 300 nm, 50 and 250 nm, 60 and 200 nm, 75 and 190 nm, 80 and 180 nm, 100 and 200 nm, 200 and 300 nm, and 150 and 250 nm. The diameter of an LNP may be determined by any method known in the art, for example, dynamic light scattering, transmission electron microscopy (TEM) or scanning electron microscopy (SEM). In some embodiments, an LNP has a diameter between 50 and 100 nm, between 70 and 100 nm, and between 80 and 100 nm. In an embodiment, an LNP has a diameter of about 90 nm. In some embodiments, an LNP described herein has a diameter greater than about 30 nm. In some embodiments, an LNP has a diameter greater than about 35 nm, about 40 nm, about 45 nm, about 50 nm, about 60 nm, about 70 nm, about 80 nm, about 90 nm, about 100 nm, about 120 nm, about 140 nm, about 160 nm, about 180 nm, about 200 nm, about 225 nm, about 49 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 250 nm, about 275 nm or about 300 nm. In an embodiment, an LNP has a diameter greater than about 70 nm. In an embodiment, an LNP has a diameter greater than about 90 nm. In an embodiment, an LNP has a diameter greater than about 180 nm. In some embodiments, a plurality of LNPs described herein has an average diameter ranging from about 40 nm to about 180 nm. In some embodiments, a plurality of LNPs described herein has an average diameter from about 50 nm to about 150 nm. In some embodiments, a plurality of LNPs described herein has an average diameter from about 50 nm to about 120 nm. In some embodiments, a plurality of LNPs described herein has an average diameter from about 60 nm to about 120 nm. In some embodiments, a plurality of LNPs has an average diameter of about 40 nm, about 45 nm, about 50 nm, about 60 nm, about 70 nm, about 80 nm, about 90 nm, about 100 nm, about 120 nm, about 140 nm, about 160 nm, about 180 nm. In some embodiments, a nanoparticle or plurality of nanoparticles described herein has an average neutral to negative surface charge of less than -100 mv, for example, less than -90 mv, - 80 mv, -70 mv, -60 mv, -50 mv, -40 mv, -30 mv, and -20 mv. In some embodiments, a nanoparticle or plurality of nanoparticles has a neutral to negative surface charge of between -100 mv and 100 mv, between -75 mv to 0, or between -50 mv and -10 mv. In some embodiments, at least 5% (e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99%) of the nanoparticles of a plurality of nanoparticles have an average neutral to negative surface charge of less than -100 mv. In some embodiments, a nanoparticle or plurality of nanoparticles has an average surface charge of between -20 mv to +20, between -10 mv and +10 mv, or between -5 mv and +5 mv at pH 7.4. LNPs that are neutral in charge have improved pharmacokinetics and biological performance compared to cationic LNPs. Making Lipid Nanoparticles (LNPs) The method of making an LNP can comprise mixing a first solution with a second solution. Mixing can be achieved using standard liquid mixing techniques, such as propellor mixing, vortexing solutions or preferably through microfluidic mixing or high efficiency T-mixing. In some embodiments, the first solution comprises a lipid or a plurality of lipids and a nucleic acid, where all components are solubilized, in water / solvent system. The solvent may be any water miscible solvent (e.g., ethanol, methanol, isopropanol, acetonitrile, dimethylformamide, dimethylsulfoxide, dioxane or tetrahydrofuran). In some embodiments, the first solution 50 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 comprises a small percentage of water or pH buffered water. The first solution may comprise up to at least 60% by volume of water, e.g., up to at least about 0.05%, 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%,45%, 50%, 55% or 60% by volume of water. In an embodiment, the first solution comprises between about 0.05% and 60% by volume of water, e.g., between about 0.05% and 50%, about 0.05% and 40%, or about 5% and 20% by volume of water. In some embodiments, the first solution comprises a single type of lipid, for example, an ionizable lipid, a phospholipid, a sterol, or a PEG-containing lipid. In some embodiments, the first solution comprises a plurality of lipids. In some embodiments, the plurality comprises an ionizable lipid, a phospholipid, a sterol, or a PEG-containing lipid. In some embodiments, the plurality of lipids comprise cholesterol, 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC),1,2-dimyristoyl- rac-glycero-3-methylpolyoxyethylene2000 (DMG-PEG2k) or α-(3’-{[1,2- di(myristyloxy)propanoxy] carbonylamino}propyl)-ω-methoxy, polyoxyethylene (PEG2000- C- DMG), and an ionizable lipid. The plurality of lipids may exist in any ratio. In an embodiment, the plurality of lipids comprises an ionizable lipid or sterol, a phospholipid, a sterol, a PEG-containing lipid of the above lipids or a combination thereof in a particular ratio (e.g., a ratio described herein). In some embodiments, the second solution is water. In some embodiments, the second solution is an aqueous buffer with a pH between 3-6 (e.g., a pH of about 3, about 4, about 5, or about 6). The second solution may comprise a load component, e.g., a nucleic acid (e.g., mRNA). The second solution may comprise a small percentage of water-miscible organic solvent. The second solution may comprise up to at least 60% by volume of at least one water miscible organic solvent, e.g., up to at least about 0.05%, 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%,10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55% , 60% or any percent therebetween by volume of at least one organic solvent (e.g., a water miscible organic solvent). In an embodiment, the second solution comprises between about 0.05% and 60% by volume of organic solvent, e.g., between about 0.05% and 50%, about 0.05% and 40%, or about 5% and 20% by volume of organic solvent (e.g., a water miscible organic solvent). The aqueous buffer solution can be an aqueous solution of citrate buffer. In some embodiments, the aqueous buffer solution is a citrate buffer solution with a pH between 4-6 (e.g., a pH of about 4, about 5, or about 6). In an embodiment, the aqueous buffer solution is a citrate buffer solution with a pH of about 6. In some embodiments, the solution comprising a mixture of the first and second solutions comprising the LNP suspension can be diluted. In some embodiments, the pH of the solution 51 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 comprising a mixture of the first and second solutions comprising the LNP suspension can be adjusted. Dilution or adjustment of the pH of the LNP suspension can be achieved with the addition of water, acid, base or aqueous buffer. In some embodiments, no dilution or adjustment of the pH of the LNP suspension is carried out. In some embodiments, both dilution and adjustment of the pH of the LNP suspension is carried out. In some embodiments, excess reagents, solvents, unencapsulated nucleic acid maybe removed from the LNP suspension by tangential flow filtration (TFF) (e.g., diafiltration). The organic solvent (e.g., ethanol) and buffer may also be removed from the LNP suspension with TFF. In some embodiments, the LNP suspension is subjected to dialysis and not TFF. In some embodiments, the LNP suspension is subjected to TFF and not dialysis. In some embodiments, the LNP suspension is subjected to both dialysis and TFF. In one aspect, the present disclosure features a method comprising treating a sample of LNPs comprising nucleic acid, with a fluid comprising a detergent (e.g., Triton X-100, or anionic detergents (such as, but not limited to, sodium dodecyl sulfate (SDS), or non-ionic detergent, such as but not limited to β-octylglucoside, or Zwittergent 3-14) for a period of time suitable to degrade the lipid layer and thereby release the encapsulated and / or entrapped nucleic acid(s). In an embodiment, the method further comprises analyzing the sample for the presence, absence, and / or amount of the released nucleic acid(s). LNP comprising ligands Some aspects of the disclosure relate to LNP comprising a ligand (also referred herein as targeting ligand) having a binding specificity for a cell surface antigen, wherein the binding of the ligand to the antigen induces the internalization of the ligand. Some embodiments relate to compositions comprising LNP comprising a ligand as described herein. LNP targeting can also accomplished by adding lipids to the formulation. For example, phosphatidylserine is known to redistribute to the external surface of the plasma membrane during apoptosis and is a molecular cue for phagocytotic cell attraction (Fadok et al. Curr Biol.2003 Aug 19;13(16):R655-7). Phosphatidylserine (PS) and phosphatidylglycerol (PG) are recognized by dendritic cells and can induce uptake and activation of dendritic cells LNP targeting can also accomplished by adding certain anionic phospholipids to the formulation (Table 3A). For example, phosphatidylserine is known to redistribute to the external surface of the plasma membrane during apoptosis and is a molecular cue for phagocytotic cell attraction (Fadok et al. Curr Biol.2003 Aug 52 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 19;13(16):R655-7). Phosphatidylserine (PS) and phosphatidylglycerol (PG) are recognized by dendritic cells and can induce uptake and activation of dendritic cells (Caronni et al., Nat Comm. 2021 April 14; 12: 2237-2253; Ischihashi et al., PLOS One 2013). Although anionic phospholipids have been used previously in the context of liposomes, their inclusion in lipidic nanoparticles that include condensed nucleic acids is unexpected since anionic headgroups may compete for binding sites of the ionizable cationic lipids with the phosphate backbone of mRNA, may inhibit intracellular escape by altering the surface charge, or may result in aggregation of LNPs during formation or storage. 53 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 Table 3A. Anionic Phospholipid Targeting Moieties ACTIVE Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 Table 3B. Phosphatidylserine Targeting Moieties O O O 9 3 - r) – – – – In some embodiments, the anionic targeting ligands are selected from the group, phosphatidylserine (PS), phoshatidylglycerol (PG), N-glutaryl-phosphatidylethanolamine (N-glu- PE), or N-succinyl-phosphatidylethanolamine (N-Suc-PE). In one embodiment, the anionic phospholipid used is phosphatidylserine. In another embodiment, the phosphatidylserine contains the L-isomer of serine. In another embodiment, the acyl chains for the phosphatidylserine are fully saturated, such as the case for dimyristoylphosphatidyl-L-serine (DMPS), dipalmitoylphosphatidyl-L-serine (DPPS), or distearoylphosphatidyl-L-serine (DSPS). In a preferred embodiment, the PS used is the L-isomer of either DPPS or DSPS. The phosphatidylserine may also contain an asymmetric acyl chain composition, for example where one acyl chain is stearic acid and another is palmitic acid. 55 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 In some embodiments, the anionic phospholipid is selected from a group other than phosphatidylserine. In some embodiments, these non PS anionic phospholipids include phosphatidylglycerol (PG), phosphatidic acid (PA), N-glutaryl-phosphatidylethanolamine (N- Glu-PE), N-succinyl-phosphatidylethanolamine (N-Suc-PE), and cardiolipin. In one embodiment, these anionic phospholipids include saturated acyl chains of 16 or 18 carbons such as distearoylphosphatidylglycerol (DSPG), dipalmitoyphosphatidylglycerol (DPPG), N-succinyl- distearoylphosphatidylethanolamine (N-Suc-DSPE), N-glutaryl-distearoylphosphatidylethanol- amine (N-Glu-DSPE), distearoylphosphatidic acid (DSPA), and cardiolipin. Table 3C. Nonphosphatidylserine anionic phospholipids O O 9 3 OH 1 O O P O OH DSPG 56 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 O O 9 3 18 1614 101 O O P - OH DSPA embodiment, the acyl chains for the phosphatidylglycerol are fully saturated, such as the case for dimyristoylphosphatidylglycerol (DMPG), dipalmitoylphosphatidylglycerol (DPPG), or distearoylphosphatidylglycerol (DSPG). In a preferred embodiment, the PG used is either DPPS or DSPS. The phosphatidylglycerol may also contain an asymmetric acyl chain composition, for example where one acyl chain is stearic acid and another is palmitic acid. In some embodiments, the salt form of phosphatidylglycerol or phosphatidylserine is highly soluble in ethanol. In some embodiments, the salt form of phosphatidylserine is highly soluble in ethanol. In some embodiments it is soluble at greater than 0.5 mg / ml, greater than 1 mg / mL, greater than 5 mg / mL, greater than 10 mg / mL, or greater than 20 mg / mL. In some embodiments the salt form of phosphatidylglycerol or phosphatidylserine is soluble is at least 0.3 mM, at least 0.4 mM, at least 0.5 mM, at least 0.6 mM, or at least 0.8 mM, as determined by a shake flask method in 200 proof ethanol, at the temperature of 22ºC of less. In some embodiments, the salt is an ammonium salt. In one embodiment, the phosphatidylserine is added to the LNP lipids in the form of ammonium or a substituted ammonium salt. Substituted ammonium salt can be mono-, di-. tri-, or tetraalkylammonium having alkyl groups with one to six, one to four, one to three, one, two, or three carbon atoms each. One or more alkyl groups can be n-alkyl, or branched alkyl groups (such as, for example, isopropyl groups), or form a ring (such as for example, cyclohexyl group). An alkyl group and the nitrogen ammonium atom may form a heterocyclic ring. The substituted ammonium salt may be also formed by an alkylenediamine. Tris(hydroxymethyl)aminomethane and triethanolamine can also be used as the amine bases to 57 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 form PS salts. In some embodiments, the amine is chosen from ammonia, dimethylamine, diethylamine, triethylamine, trimethylamine, 2-(dimethyamino)ethanol, diethanolamine, 2- (diethyamino)ethanol, ethanolamine, ethylenediamine, N-methyl-glucamine, imidazole, histidine, lysine, arginine, 4-(2-hydroxyethyl)-morpholine, piperazine, 1-(2-hydroxyethyl)-pyrrolidine, triethanolamine, and tromethamine (tris(hydroxymethyl)aminomethane), In some embodiments, this targeting lipid is an ammonium salt of DPPS. Table 3D. Ammonium and sodium salt forms of dipalmitoyl- or distearoyl-phosphatidylserine. or any method known in the art may be used. In one embodiment, a sodium salt of phosphatidylserine (PS) is dissolved in a monophase system of chloroform, methanol, and water, containing a chloride salt of ammonium or substituted ammonium (a Bligh-Dyer monophase), and the system is brought 58 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 to the two-phase state by adding extra methanol and / or water containing the ammonium or substituted ammonium chloride. The chloroform-rich phase, containing the PS, is separated, and the process is repeated. Finally, the chloroform-rich phase is washed with water to remove excess chloride, and the ammonium (substituted ammonium) salt of PS is obtained by evaporation of the chloroform-rich phase. Optionally, the obtained ammonium or substituted ammonium salt of PS is vacuum dried or dissolved in cyclohexane and lyophilized. In another embodiment, the PS as a sodium or potassium salt is dissolved in a water-immiscible organic solvent, such as chloroform or a chloroform-methanol mixture, and washed with diluted aqueous solution of an acid, such as HCl, to obtain a free acid form of the PS, which is then neutralized with ammonium hydroxide or substituted amine in free base form. In yet another embodiment, the organic solution of PS as a sodium or potassium salt is treated with a cation-exchange resin in the ammonium of substituted ammonium form. In yet another embodiment, the PS is prepared in the form of a calcium or magnesium salt and treated with ammonium or substituted ammonium salt of a chelator, such as EDTA, or with ammonium or substituted ammonium phosphate, in the presence of an organic solvent, causing displacement of calcium or magnesium ion in the form of a chelate or a yet less soluble phosphate, which is separated, e.g., by filtration, while ammonium or substituted ammonium salt of PS is left in the organic (e.g., ethanol) solution. In one embodiment, PS or PG are added to the LNP lipid formulation at a concentration between about 0.1 mol% to about 20 mol%, about 0.1 mol% to about 10 mol%, about 0.1 mol% to about 5 mol%, about 0.5 mol% to about 20 mol%, about 0.5 mol% to about 10 mol%, about 0.5 mol% to about 5 mol%, about 1 mol% to about 20 mol%, about 1 mol% to about 10 mol%, or about 1 mol% to about 5 mol%, of the total lipid content of the LNP. In one embodiment, the PS is added to the LNP lipid formulation at a concentration between about 1 mol% to about 20 mol%, about 2.5 mol% to about 10 mol%, about 3 mol% to about 9 mol%, or about 4 mol% to about 8 mol%, of the total lipid content of the LNP. In one embodiment the PS or PG lipid is included in the LNP composition comprising ionizable cationic lipids known in the art, including DODAP, AKG-OA-DM2, O-11769, DLin- MC3-DMA, DLin-KC2-DMA, DLin-KC3-DMA, ALC-0315, and SM-102. In another embodiment the PS lipid is included in the LNP composition comprising ICLs of Formula I, II, III, IV-B, V-A-1, combinations thereof or pharmaceutically salts thereof. In 59 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 another embodiment the PS lipid is included in the LNP composition using N / P ratios between 3 and 8, between 4 and 7, or between 5 and 6. In some aspects, a method of delivering a nucleic acid to a cell is provided, the method comprising: contacting the cell with a composition comprising an LNP comprising a ligand (also referred herein as targeting ligand) having a binding specificity for a cell surface antigen, wherein the binding of the ligand to the antigen induces the internalization of the ligand. In some embodiments, the targeting ligand can be, but is not limited to, an internalizing antibody, or a fragment thereof, a small molecule conjugates or gylcoconjugates. In some embodiments, the binding of the targeting ligand to a specific cell surface antigen induces the internalization of the LNP with the targeting ligand attached by a cell expressing at least 100,000 or at least 1,000,000 molecules of the antigen when contacted and incubated with the cell under internalizing conditions. Table 4A. Exemplary dialkyl and branched ionizable cationic lipids ACTIVE Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 AKG-KC4-OA AKG-KC4-OA(S) 61 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 Table 4B. Exemplary dialkyl and branched ionizable cationic lipids (Continued) Compositions In some embodiments, a lipidic nanoparticle composition comprises lipids and nucleic acids, the lipidic nanoparticles comprising a compound of Formula I, II, III, IV-B, V-A-1, combinations thereof or pharmaceutically acceptable salts thereof. Other aspects of the disclosure relate to the use of these ionizable lipids or lipidic nanoparticles compositions comprising ionizable lipids in vaccines for the prevention of infectious diseases or cancer. In some embodiments, the infectious disease can be a bacterial or a viral infection. In some embodiments, the compositions described herein can be used to prevent infections related to tuberculosis, HIV / AIDS, malaria, or coronavirus-related infections such as COVID-19. In other embodiments, the infection is influenza, hepatitis B, hepatitis C, Dengue, human papillomavirus (HPV), norovirus, mumps, measles, Meningococcal disease, pneuomococcal disease, polio, rotovirus, respiratory syncytial virus (RSV), rubella, shingles / herpes zoster virus, tetanus, or whooping cough. In some embodiments, the compounds and compositions described herein may promote efficient uptake and transfection of target cells, including tissue macrophages and dendritic cells. The efficient delivery nucleic acids coding for antigen specific for infectious viruses or bacteria, and subsequent presentation of that antigen to elicit the desired immune response to protect against corresponding infections is a result. In some embodiments, the nucleic acid can be a synthetic nucleic acid (e.g., engineered codon optimized mRNA) encoding an epitope of coronavirus such 62 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 as SARS-CoV, MERS-CoV or SARS-CoV-2. In some embodiments the nucleic acid can be a synthetic nucleic acid (e.g. engineered codon optimized mRNA) encoding the S-protein (spike protein) or a fragment thereof of coronavirus such as SARS-CoV, MERS-CoV or SARS-CoV-2. In some embodiments, the vaccine is used for the prevention mycobacterium infections. In some embodiments, the vaccine can be used for the prevention of tuberculosis, nontuberculous mycobacteria (NTM), nontuberculosis lung disease, leprosy, mycobacterium avium-intracellulare, mycobacterium kansasii, mycobacterium marinum, mycobacterium ulcerans, mycobacterium chelonae, mycobacterium fortuitum, mycobacterium abscessus and other infectious diseases such as those caused by coronaviruses (SARS-CoV-2, Covid-19; SARS-CoV, SARS; MERS-CoV; HCoV-229E; HCoV-NL63; HCoV-OC43; HCoV-HKU1), chikungunya, dengue, diphtheria, Ebola, EV-D68, influenza (flu), hepatitis viruses (including HAV, HBV, HCV, HDV, HEV and GB virus C), Haemophilus influenzae type B (Hib), Hendra virus, HIV / AIDS, human metapneumovirus (hMPV), Human Ppapillomaviruses (HPV), Lassa, Lyme, malaria, Marburg, measles, meningococcal disease, mumps, Nipah virus, norovirus, parainfluenza virus (PIV), plague, pneumococcal disease, polio, respiratory syncytial virus (RSV), Rocky Mountain spotted fever, rotavirus, rubella (German Measles), varicella zoster virus (chickenpox, shingles), smallpox, tetanus (Lockjaw), West Nile, whooping cough (Pertussis) and zika In some embodiments, the composition further comprises a pharmaceutical excipient. In some embodiments, the lipidic nanoparticles are in an aqueous medium. In some embodiments, the nucleic acid is entrapped in the lipidic nanoparticle with an ionizable cationic lipid compound provided herein or combinations thereof, wherein the nucleic acid is either RNA or DNA. In some embodiments, the nucleic acid is mRNA. In some embodiments, the nucleic acid is siRNA. In some embodiments, the nucleic acid is DNA. In some embodiments, the lipidic nanoparticle comprises a membrane comprising phosphatidylcholine and a sterol. In some embodiments, the sterol is cholesterol. In other embodiments, the sterol is beta-sitosterol. In some embodiments, the lipidic nanoparticle comprises a membrane comprising phosphatidylcholine, ionizable cationic lipid (ICL). In some embodiments, the ICL have a structure of Formula I, and cholesterol, wherein the membrane separates the inside of the lipidic nanoparticles from the aqueous medium. In some embodiment, the ICL have a structure as shown in Table 1A and Table 2. In some embodiment, the ICL have a 63 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 structure as shown in Table 1B. In some embodiments, the phosphatidylcholine is distearoylphosphatidylcholine (DSPC) or hydrogenated soy phosphatidylcholine (HSPC). In some embodiments, the ionizable cationic lipid to cholesterol molar ratios is from about 65:35 to 40:60. In some embodiments, the ICL to cholesterol molar ratio is from about 60:40 to about 45:55. In some embodiments, the phosphatidylcholine to cholesterol molar ratio is from about 1:5 to about 1:2. In some embodiments, the membrane further comprises a polymer-conjugated lipid. In some embodiments, the lipidic nanoparticle comprises ICL, DSPC, cholesterol and polymer-conjugated lipid in an about 49.5:10.3:39.6:2.5 molar ratio. In some embodiments, the polymer-conjugated lipid is PEG(2000)-dimyristoylglycerol (PEG-DMG) or PEG(Mol. weight 2,000)-dimyristoylphosphatidylethanolamine (PEG-DMPE). The compositions of this disclosure may be administered by various routes, for example, to effect systemic delivery via intravenous, parenteral, intraperitoneal, or topical routes. The compositions may be administered intravenously, subcutaneously, or intraperitoneally to a subject. In some embodiments, the disclosure provides methods for in vivo delivery of nucleic acids to a subject. In some embodiments, the composition is a liquid pharmaceutical formulation for parenteral administration. In some embodiments, the composition is a liquid pharmaceutical formulation for subcutaneous, intramuscular, or intradermal administration. In some embodiments, the composition is in the form of a lyophilized powder, that is subsequently reconstituted with aqueous medium prior to administration. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I): ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 wherein R1is , are and n is an integer equal to 2, 3 or 4. wherein a and b of the two R1hydrocarbon chains are the same or different, or one of the two R1 hydrocarbon chains is a saturated C12-C18 alkyl. In some embodiments, ionizable cationic lipid compositions are provided. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I-A): , R2and R3are each independently (C1-C4) alkyl optionally substituted with hydroxyl; and n is an integer equal to 2, 3 or 4. In some embodiments, a and b of the two R1 hydrocarbon chains are the same. In some embodiments, a and b of the two R1hydrocarbon chains are different. In some embodiments, one of the two R1hydrocarbon chains is a saturated C12-C18alkyl. In some embodiments, ionizable cationic lipid compositions are provided. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I-A): 65 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 , 4; R2 and R3 are each methyl; and n is an integer equal to 3. In some embodiments, a and b of the two R1hydrocarbon chains are the same. In some embodiments, a and b of the two R1hydrocarbon chains are different. In some embodiments, one of the two R1 hydrocarbon chains is a saturated C12-C18 alkyl. In some embodiments, ionizable cationic lipid compositions are provided. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I-A): wherein R1 is a saturated C15-C18 hydrocarbon chain, R2 and R3 are each methyl; and n is an integer equal to 3. In some embodiments, a and b of the two R1 hydrocarbon chains are the same. In some embodiments, a and b of the two R1 hydrocarbon chains are different. 66 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 In some embodiments, one of the two R1hydrocarbon chains is a saturated C12-C18alkyl. Methods of use Targeting of dendritic cells Dendritic cells (DCs) are specialized antigen-presenting cells that play a central role in initiating and regulating adaptive immunity. Owing to their potent antigen (Ag) presentation capacity and ability to generate distinct T-cell responses, efficient and specific delivery of Ags to DCs is the cornerstone for generating Ag-specific effector and memory cells against tumors or pathogens. Dendritic cells can be generated from human blood monocytes by adding granulocyte- macrophage colony-stimulating factor (GM-CSF), IL-4, and IFN-gamma to differentiate monocyte-derived DC in vitro. Cells in culture exhibit both dendritic and veiled morphologies, the former being adherent, and the latter suspended. Phenotypically, they are CD1a- / dim, CD11a+, CD11b++, CD11c+, CD14dim / -, CD16a- / dim, CD18+, CD32dim / -, CD33+, CD40+, CD45R0+, CD50+, CD54+, CD64- / dim, CD68+, CD71+, CD80dim, CD86+ / ++, MHC class I++ / , HLA- DR++ / , HLA-DP+, and HLA-DQ (Geiseler et al. Dev Immunol.1998;6(1-2):25-39). Alternatively, human primary blood dendritic cell lines have been developed and are commercially available from Creative Biolabs. CD8+ T cells can produce IL2, IFN-γ, and TNF, cytokines that are known to have critical functions during mycobacterium tuberculosis infection. Importantly, CD8+ T cells have cytolytic functions to kill mycobacterium tuberculosis -infected cells via granule-mediated function (via perforin, granzymes, and granulysin) or Fas-Fas ligand interaction to induce apoptosis. In humans, CD8+ T cell can produce granulysin, which can kill mycobacterium tuberculosis directly. Therefore, it is anticipated that antigen generating mRNA LNPs delivered to DC will stimulate a CD8+ T cell response to fight against mycobacterium tuberculosis infection. CD8+ T cells are able to recognize M. tuberculosis specific antigens (as peptides) presented by classical and non-classical MHC molecules. Classically restricted CD8+ T cells have been identified that recognize antigens presented by antigen presenting cells in the context of classical MHC Ia (HLA-A, -B, -C) molecules. Non-classically restricted CD8+ T cells include those CD8+ T cells that are capable of recognizing Mg antigen in the context of HLA-E molecules (non-MHC 1a), glycolipids associated with group 1 CD1 molecules and MHC I-related molecules (MR1) such as mucosal associated invariant T cells (MAIT). Finally, γδ T cells represent a separate population 67 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 of CD8 (and CD4) T cells that have both innate and adaptive functions in response to mycobacterium tuberculosis infection. CD8+ T cells have been shown to play direct functions in response to mycobacterium tuberculosis infection but they also play important roles in orchestrating many different functions in the overall host immune response (e.g., interaction to provide optimal CD4 T cell function) In one embodiment, LNPs can be added to cultured human dendritic cells at an appropriate concentration, (e.g.1-5 µg / mL mRNA). After some time to allow for cellular uptake and antigen expression, human T cells (HemaCare) can be added, and the cell culture media is sampled at various times for INF-γ by Elisa (R&D Systems, DIF50C). Alternatively, the cells can be analyzed by flow cytometry for CD8+ marker or intracellular INFγ production (PE anti-human IFN- γ antibody, Biolegend). In one embodiment, LNPs can be administered into a subject at a dose of 0.01-5 mg / kg mRNA by any route of administration outlined above. According to some embodiments, a proportion of LNPs are taken up DC cells, while most will accumulate in the liver and spleen. The DC cells can express the antigenic peptide, process it for MHC I presentation and travel to the lymph node for presentation to naïve T cells inducing an education of memory T-cells towards the antigen. In one embodiment, LNPs that have been modified with a targeting ligand such as anti- DEC205-PEG-DSPE can be administered into a subject at a dose of 0.01-5 mg / kg mRNA. According to some embodiments, a higher proportion of LNPs can be taken up DC cells, allowing for increased production of antigenic peptide compared to non-targeted LNP and a more efficient vaccination against the pathogen. Additional targeting ligands for dendritic cells include, but are not limited to, CLEC9A, CLEC4A, XCR1, CD141, and HLD-DR. For example, assessing the CD8+ reactivity to the in vivo produced antigen could be accomplished by measuring INFγ plasma levels by species specific IFN-gamma Quantikine ELISA Kits from R&D Systems. In some embodiments, LNP compositions provide desirable pharmacokinetic properties such as extended plasma half-life and stable encapsulation of mRNA. The plasma half-life can be measured as the percentage of the injected dose (ID) remaining in blood after 6 or 24 hours following injection intravenously in immunocompetent mice. The stability of the encapsulation of mRNA over 24 hours in plasma can be determined by changes in the mRNA-to-lipid ratio (mRNA / L ratio) following iv administration in mice. In some embodiments, the percentage of 68 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 encapsulated mRNA remaining in blood is greater than 20 %, preferably greater than 30 %, and most preferably greater than 40 % of the injected dose at 6 hours. The percent retained in blood after 24 h is preferably greater than 10 %, and more preferably greater than 20 % of the injected dose. Disclosed herein are methods for preventing mycobacteria infection, such as Mycobacterium tuberculosis, or gram-positive bacteria, such as methicillin-resistant Staphylococcus aureus (MRSA). Additional mycobacteria and gram positive bacteria include, but are not limited to, Mycobacterium avium complex, Mycobacterium leprae, Mycobacterium gordonae, Mycobacterium abscessus, Mycobacterium abscessus, Mycobacterium mucogenicum, streptococci, vancomycin-resistant enterococci (VRE), Staphylococcus pneumoniae, Enterococcus faecium, Streptococcus agalactiae, Streptococcus pneumoniae, Streptococcus pyogenes, the viridans group streptococci, Listeria monocytogenes, Nocardia, and Corynebacterium. Administration of a vaccine for inducing a second immune response may provide MHC class II - presented epitopes that are capable of eliciting a CD4 + helper T cell response against cells expressing antigens from which the MHC presented epitopes are derived. Alternatively or additionally, administration of a vaccine for inducing a second immune response may provide MHC class I - presented epitopes that are capable of eliciting a CD8 + T cell response against cells expressing antigens from which the MHC presented epitopes are derived. Furthermore, administration of a vaccine for inducing a second immune response may provide one or more neo - epitopes (including known neo epitopes) as well as one or more epitopes not containing cancer specific somatic mutations but being expressed by cancer cells and preferably inducing an immune response against cancer cells, preferably a cancer specific immune response. In one embodiment, administration of a vaccine for inducing a second immune response provides neo - epitopes that are MHC class Il - presented epitopes and / or are capable of eliciting a CD4 + helper T cell response against cells expressing antigens from which the MHC presented epitopes are derived as well as epitopes not containing cancer - specific somatic mutations that are MHC class I - presented epitopes and / or are capable of eliciting a CD8 + T cell response against cells expressing antigens from which the MHC presented epitopes are derived. In one embodiment, the epitopes do not contain cancer - specific somatic mutations. 69 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 A "cellular immune response”, a "cellular response”, a “cellular response against an antigen” or a similar term is meant to include a cellular response directed to cells characterized by presentation of an antigen with class I or class II MHC. The cellular response relates to cells called T cells or T - lymphocytes which act as either “helper cells” or “killer cells”. The helper T cells (also termed CD4 + T cells ) play a central role by regulating the immune response and the killer cells (also termed cytotoxic T cells , cytolytic T cells , CD8 + T cells or CTLS ) kill diseased cells such as cancer cells, preventing the production of more diseased cells. In preferred embodiments, the present disclosure involves the stimulation of an anti-Mycobacterium tuberculosis CTL response against the mycobacterium expressing one or more expressed antigens and preferably presenting such expressed antigens with class I MHC. An “antigen” according to the disclosure covers any substance that will elicit an immune response. In particular, an “antigen” relates to any substance, preferably a peptide or protein, that reacts specifically with antibodies or T-lymphocytes (T cells). As used herein, the term “antigen” comprises any molecule which comprises at least one epitope. Preferably, an antigen in the context of the present disclosure is a molecule which, optionally after processing, induces an immune reaction, which is preferably specific for the antigen (including cells expressing the antigen). According to the present disclosure, any suitable antigen may be used, which is a candidate for an immune reaction, wherein the immune reaction is preferably a cellular immune reaction. In the context of the embodiments of the present disclosure, the antigen is preferably presented by a cell, preferably by an antigen presenting cell which includes a diseased cell, in particular a cancer cell, in the context of MHC molecules, which results in an immune reaction against the antigen. An antigen is preferably a product which corresponds to or is derived from a naturally occurring antigen. Such naturally occurring antigens may include tumor antigens. As used herein, an “antigen peptide” relates to a portion or fragment of an antigen which is capable of stimulating an immune response, preferably a cellular response against the antigen or cells characterized by expression of the antigen and preferably by presentation of the antigen such as diseased cells, in particular cancer cells. Preferably, an antigen peptide is capable of stimulating a cellular response against a cell characterized by presentation of an antigen with class I MHC and preferably is capable of stimulating an antigen - responsive cytotoxic T - lymphocyte (CTL). Preferably, the antigen peptides according to the disclosure are MHC class I and / or class II presented peptides or can be processed to produce MHC class I and / or class II presented 70 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 peptides. Preferably, the antigen peptides comprise an amino acid sequence substantially corresponding to the amino acid sequence of a fragment of an antigen. Preferably, said fragment of an antigen is an MHC class I and / or class II presented peptide. Preferably, an antigen peptide according to the disclosure comprises an amino acid sequence substantially corresponding to the amino acid sequence of such fragment and is processed to produce such fragment, i.e., an MHC class I and / or class II presented peptide derived from an antigen. If a peptide is to be presented directly, i.e., without processing, in particular without cleavage, it has a length which is suitable for binding to an MHC molecule, in particular a class I MHC molecule, and preferably is 7 - 20 amino acids in length, more preferably 7 - 12 amino acids in length, more preferably 8 - 11 amino acids in length, in particular 9 or 10 amino acids in length. The main types of professional antigen - presenting cells are dendritic cells, which have the broadest range of antigen presentation, and are probably the most important antigen - presenting cells, macrophages, B - cells, and certain activated epithelial cells. Dendritic cells (DCs ) are leukocyte populations that present antigens captured in peripheral tissues to T cells via both MHC class II and I antigen presentation pathways . It is well known that dendritic cells are potent inducers of immune responses and the activation of these cells is a critical step for the induction of antitumoral immunity. Dendritic cells are conveniently categorized as “immature” and “mature” cells, which can be used as a simple way to discriminate between two well characterized phenotypes. However, this nomenclature should not be construed to exclude all possible intermediate stages of differentiation. Immature dendritic cells are characterized as anti gen presenting cells with a high capacity for antigen uptake and processing, which correlates with the high expression of Fcγ receptor and mannose receptor. The mature phenotype is typically characterized by a lower expression of these markers, but a high expression of cell surface molecules responsible for T cell activation such as class I and class II MHC, adhesion molecules (e.g. CD54 and CD11) and costimulatory molecules (e.g., CD40, CD80, CD86 and 4 - 1 BB). Dendritic cell maturation is referred to as the status of dendritic cell activation at which such antigen - presenting dendritic cells lead to T cell priming, while presentation by immature dendritic cells results in tolerance. Dendritic cell maturation is chiefly caused by biomolecules with microbial features detected by innate receptors (bacterial DNA, viral RNA, endotoxin, etc), pro-inflammatory cytokines (TNF, IL - 1, IFNs), ligation of CD40 on the dendritic cell surface by CD4OL, and substances 71 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 released from cells undergoing stressful cell death. The dendritic cells can be derived by culturing bone marrow cells in vitro with cytokines, such as granulocyte - macrophage colony - stimulating factor (GM CSF) and tumor necrosis factor alpha. Non - professional antigen-presenting cells do not constitutively express the MHC class II proteins required for interaction with naive T cells; these are expressed only upon stimulation of the non - professional antigen-presenting cells by certain cytokines such as IFNγ. "Antigen presenting cells” can be loaded with MHC class I presented peptides by transducing the cells with nucleic acid, preferably mRNA, encoding a peptide or polypeptide comprising the peptide to be presented, e.g. a nucleic acid encoding the antigen. In some embodiments, a pharmaceutical composition comprising a gene delivery vehicle that targets a dendritic or other antigen presenting cell may be administered to a patient, resulting in transfection that occurs in vivo. As used herein, a “nucleic acid” is a deoxyribonucleic acid (DNA) or ribonucleic acid (RNA), more preferably RNA, most preferably in vitro transcribed RNA (IVT RNA) or synthetic RNA. Nucleic acids include according to the disclosure genomic DNA, cDNA, mRNA, recombinantly produced and chemically synthesized molecules. According to the disclosure, a nucleic acid may be present as a single - stranded or double - stranded and linear or covalently circularly closed molecule. A nucleic acid can, according to the disclosure, be isolated. The term “isolated nucleic acid” means, according to the disclosure, that the nucleic acid (i) was amplified in vitro, for example via polymerase chain reaction (PCR), (ii) was produced recombinantly by cloning, (iii) was purified, for example, by cleavage and separation by gel electrophoresis, or (iv) was synthesized, for example, by chemical synthesis. A nucleic can be employed for introduction into, i.e. transfection of cells, in particular, in the form of RNA which can be prepared by in vitro transcription from a DNA template. The RNA can moreover be modified before application by stabilizing sequences, capping, and polyadenylation. As used herein, the term “RNA” relates to a molecule which comprises ribonucleotide residues and preferably being entirely or substantially composed of ribonucleotide residues. “Ribonucleotide” relates to a nucleotide with a hydroxyl group at the 2 ' - position of a B-D- ribofuranosyl group. The term “RNA” comprises double - stranded RNA, single - stranded RNA, isolated RNA such as partially or completely purified RNA, essentially pure RNA, synthetic RNA, and recombinantly generated RNA such as modified RNA which differs from naturally occurring RNA by addition, deletion, substitution and / or alteration of one or more nucleotides. Such 72 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 alterations can include addition of non - nucleotide material, such as to the end(s) of a RNA or internally, for example at one or more nucleotides of the RNA. Nucleotides in RNA molecules can also comprise non - standard nucleotides, such as non-naturally occurring nucleotides or chemically synthesized nucleotides or deoxynucleotides. These altered RNAs can be referred to as analogs or analogs of naturally - occurring RNA. As used herein, the term “RNA” includes and preferably relates to “mRNA”. The term "mRNA” means "messenger - RNA” and relates to a “transcript” which is generated by using a DNA template and encodes a peptide or polypeptide. Typically, an mRNA comprises a 5 ' - UTR, a protein coding region, and a 3 ' - UTR. mRNA only possesses limited half - life in cells and in vitro. In the context of the present disclosure, mRNA may be generated by in vitro transcription from a DNA template. The term “modification” in the context of the RNA used in the present disclosure includes any modification of an RNA which is not naturally present in said RNA. In one embodiment of the disclosure, the RNA used according to the disclosure does not have uncapped 5'-triphosphates. Removal of such uncapped 5'- triphosphates can be achieved by treating RNA with a phosphatase. The RNA according to the disclosure may have modified ribonucleotides in order to increase its stability and / or decrease cytotoxicity. For example, in one embodiment, in the RNA used according to the disclosure 5-methylcytidine is substituted partially or completely, preferably completely, for cytidine. Alternatively or additionally, in one embodiment, in the RNA used according to the disclosure pseudouridine is substituted partially or completely, preferably completely, for uridine. In some embodiments, the term “modification” relates to providing an RNA with a 5 - cap or 5' - cap analog. The term “5 - cap” refers to a cap structure found on the 5 ' - end of an mRNA molecule and generally consists of a guanosine nucleotide connected to the mRNA via an unusual 5' to 5 triphosphate linkage. In one embodiment, this guanosine is methylated at the 7 - position. The term "conventional 5' - cap” refers to a naturally occurring RNA 5 '-cap, preferably to the 7 - methylguanosine cap (m'G). as used herein, the term “5' - cap” includes a 5' - cap analog that resembles the RNA cap structure and is modified to possess the ability to stabilize RNA and / or enhance translation of RNA if attached thereto, preferably in vivo and / or in a cell. According to the disclosure, the stability and translation efficiency of RNA may be modified as required. For example, RNA may be stabilized and its translation increased by one or more modifications having a stabilizing effects and / or increasing translation efficiency of RNA. 73 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 Such modifications are described, for example, in PCT / EP2006 / 009448 incorporated herein by reference. In order to increase expression of the RNA used according to aspects of the disclosure, it may be modified within the coding region, i.e. the sequence encoding the expressed peptide or protein, preferably without altering the sequence of the expressed peptide or protein, so as to increase the GC content to increase mRNA stability and to perform a codon optimization and, thus, enhance translation in cells. Aspects of the disclosure relate to a method of preventing a bacterial or viral infection, the method comprising administering to a subject in need thereof an effective amount of the composition provided herein to elicit an immune response. Aspects of the disclosure provide methods of vaccinating a subject comprising administering to the subject a single dosage of the compositions described herein comprising a nucleic acid (e.g. mRNA) encoding a polypeptide in an effective amount to vaccinate the subject. In some embodiments, the nucleic acid is formulated within a cationic lipidic nanoparticle. In some embodiments, the lipidic nanoparticle composition is administered as a single injection. In some embodiments, the bacterial infection is Mycobacterium tuberculosis infection. In some embodiments, the viral infection is a coronavirus. In some embodiments, the coronavirus is SARS-CoV, MERS-CoV or SARS-CoV-2 In some embodiments, the viral infection is HIV / AIDS. In some embodiments, the lipidic nanoparticle is administered parenterally. In general, administration to a patient by intradermal injection is possible. However, injection may also be carried out intranodally into a lymph node (Maloy et al. (2001), Proc Natl Acad Sci USA 98:3299-3033). The resulting cells present the complex of interest and are recognized by autologous cytotoxic T lymphocytes which then propagate. In some embodiments, the compositions are administered by inhalation. In some embodiments, the composition is formulated as nasal spray, and / or aerosol. Actual dosage levels of the active agents in the pharmaceutical compositions disclosed herein may be varied so as to obtain an amount of the active agent which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient. “Parenteral” as used herein in the context of administration means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, 74 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrasternal injection and infusion. The phrases “parenteral administration” and “administered parenterally” as used herein refer to modes of administration other than enteral (i.e., via the digestive tract) and topical administration, usually by injection or infusion, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, inhalation, subcapsular, subarachnoid, respiratory mucosal, intraspinal, epidural and intrasternal injection and infusion. Intravenous injection and infusion are often (but not exclusively) used for liposomal drug administration. Dosage regimens can be adjusted to provide the optimum desired response (e.g., a therapeutic response). For example, one or more doses may be administered over time or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. In some embodiments, the dose comprises between 0.01 to 5 mg / kg of nucleic acid. In some embodiments, the dose comprises between 0.01 to 5 mg / kg of mRNA. In some embodiments, the dose comprises between 0.01 to 3 mg / kg of nucleic acid. In some embodiments, the dose comprises between 0.01 to 3 mg / kg of mRNA. In some embodiments, the dose comprises between 0.01 to 1 mg / kg of nucleic acid. In some embodiments, the dose comprises between 0.01 to 1 mg / kg of mRNA. In some embodiments, the dose comprises between 0.01 to 0.5 mg / kg of nucleic acid. In some embodiments, the dose comprises between 0.01 to 0.5 mg / kg of mRNA. In some embodiments, the dose comprises between 0.01 to 1 mg / kg of mRNA. In some embodiments, the dose comprises between 0.01 to 0.1 mg / kg of nucleic acid. In some embodiments, the dose comprises between 0.01 to 0.05 mg / kg of mRNA. In some embodiments, the dose comprises between 0.01 to 0.1 mg / kg of nucleic acid. In some embodiments, the dose comprises between 0.01 to 0.05 mg / kg of mRNA. The dosage of the compounds and / or of their pharmaceutically acceptable salts or the LNPs comprising the compounds and / or of their pharmaceutically acceptable salts may vary within wide limits and should naturally be adjusted, in each particular case, to the individual conditions and to the pathogenic agent to be controlled. 75 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 In some embodiments, ionizable cationic lipids (ICLs) are provided. Cationic lipids are engineered with improved stability to oxidative degradation while in storage, while retaining high transfection activity or potency in cells. Aspects of the disclosure are based in part on the discovery that LNP compositions comprising mRNA and certain ionizable cationic lipids (ICL) enhanced expression of the mRNA in human dendridic cells. In some embodiments, LNP compositions comprise a targeting ligand directed against cell surface receptors to target lipid nanoparticles in a highly specific manner, including to dendritic cells. In some embodiments, the LNP composition comprises a phosphatidyl-L-serine compound as a targeting ligand, such as dipalmitoylphosphatidyl-L-serine (DPPS), or distearoylphosphatidyl- L-serine (DSPS). In some embodiments, the LNP composition comprises a phosphatidyl-L-serine compound as a targeting ligand and an anionic phospholipid. In some embodiments, the LNP composition comprises a phosphatidylglycerol-containing compound as a targeting ligand such as distearoylphosphatidylglycerol (DSPG) or dipalmitoyphosphatidylglycerol (DPPG), for enhancing expression in human dendritic cells. In some embodiments, LNP compositions comprise both a phosphatidyl-L-serine compound as a targeting ligand, and distearoylphosphatidylcholine (DSPC) as the second phospholipid. In some embodiments, LNP compositions comprise both a phosphatidyl-L-serine compound as a targeting ligand, and distearoylphosphatidylcholine (DSPC) as the second phospholipid without dipalmitoylphosphatidylcholine (DPPC). Aspects of the disclosure are based in part on the discovery that selection of certain cationic ionizable lipids can enhance the transfection of human dendritic cells. For example, the KC3 cationic ionic lipids were more active in transfecting human dendritic cells in LNP compositions than either the KC2 or diacyl ionizable lipids (UO series). Among the LNP compositions comprising KC3 ionizable cationic lipids, those LNP compositions with ionizable cationic lipids having monounsaturated alkyl chains were unexpectedly both more active and more stable to oxidative degradation than those containing those with the dilinoleyl alkyl chains. In addition, we observed reduced transfection in human dendritic cells activity in LNP compositions comprising the monounsaturated lipids, such as for the diacyl ionizable lipids (UO-series). In some embodiments, certain salts of the phosphatidylserine targeting lipids are provided. For example, in some embodiments, the phosphatidylserine targeting lipids can be provided as an 76 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 ammonium salt of DPPS having improved biophysical properties and higher solubility in the presence of ethanol, a preferred solvent for preparation of LNPs. The sodium salts of DSPS or DSPS were insoluble in ethanol and required both the presence of methanol and heating to allow for their formation, as did the ammonium salt of DSPS. It is contemplated that the other ammonium salts of phosphatidylserine will give rise to the same advantages in solubility and biophysical properties. In some embodiments, ionizable cationic lipid compositions useful in the preparation of liposomal nanoparticle (LNP) compositions are provided. In some embodiments, liposomal compositions are provided comprising an ionizable cationic lipid having (a) a pair of linear C16 or C18 hydrocarbon chains each comprising a single unsaturated alkenyl double bond within each polyene hydrocarbon chain, covalently bound to a head group comprising a dialkyl amino alkyl group. In some embodiments, the head group of the ionizable cationic lipid has a dialkyl amino group having a pKa of about 6.3 -7.5. In some embodiments, the head group of the ionizable cationic lipid comprises a heterocyclyl or alkyl portion covalently bound to the dialkyl amino group. In some embodiments, the head group of the ionizable cationic lipid optionally further comprises a phosphate group. In some embodiments, each lipid tail of the ionizable cationic lipid compound is identical, and each lipid tail has a total of one olefin with a total length of 15, 16, 17 or 18 carbons. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I): , R2and R3are each independently (C1-C4) alkyl optionally substituted with hydroxyl; and 77 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 n is an integer equal to 2, 3 or 4. In some embodiments, ionizable cationic lipid compositions are provided. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I), wherein a is 0 or 1; b is 1, 2, 3 or 4, provided the sum a+b is 1, 2, 3 or 4; R2and R3are each methyl; and n is an integer equal to 3. In some embodiments, ionizable cationic lipid compositions are provided. In some embodiments, a lipid nanoparticle (LNP) composition comprises an ionizable cationic lipid having the chemical structure of Formula (I-A): , R2 and R3 are each methyl; and n is an integer equal to 3. In some embodiments, a LNP composition comprises an ionizable cationic lipid comprises a pair of identical, lipid hydrocarbon tails having a total of 15, 16, 17 or 18 carbons and comprising a single olefin group, or a pair of olefin groups. In some embodiments, a LNP composition comprises an ionizable cationic lipid selected from the group consisting of: 78 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 N DLIN-KC3-DMA O the ionizable cationic lipid is KC3-C15 (C8:1). In some embodiments, the ionizable cationic lipid is KC3-C16 (C8:1). In some embodiments, the ionizable cationic lipid is KC3-C17 (C8:1). In some embodiments, the ionizable cationic lipid is KC3-C18 (C8:1). In some embodiments, the ionizable cationic lipid is KC3-15. In some embodiments, the ionizable cationic lipid is KC3-16. In some embodiments, the ionizable cationic lipid is KC3-17. In some embodiments, the ionizable cationic lipid is KC3-18. 79 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 The salt form of the targeting lipid can influence it’s solubility in alcohol containing solvents used in the preparation of lipid nanoparticles. In some embodiments, ionizable cationic lipid compositions are provided. In some embodiments, a lipid nanoparticle (LNP) composition comprises a nucleic acid; an ionizable lipid disclosed herein; a sterol; one or more phospholipids comprising a phosphatidylserine (PS) lipid; and optionally further comprising a conjugated lipid. In some embodiments, a lipid nanoparticle (LNP) composition comprises a mRNA nucleic acid; an ionizable lipid disclosed herein; cholesterol;.one or more phospholipids selected from the group consisting of: DSPC, DPPC and DOPC; and a PS lipid selected from the group consisting of: DPPS, DSPS and DOPS; and optionally further comprising a conjugated lipid comprising PEG. . In some . In some embodiments, the ionizable lipid is AKG-UO-1B: In some 80 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 . In some . In some . In some . some 7, AKG- UO-8, AKG-UO-9, or AKG-UO-10: 81 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 embodiments, a LNP composition is prepared using a sodium or ammonium salt of an anionic phospholipid. In some embodiments, the anionic phospholipid salt is a compound of Formula (V- A-1), having the chemical structure: , wherein 82 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 X+is an ammonium (NH4+) or sodium (Na+) cation; and a is 14, 15 or 16. In some embodiments, the anionic phospholipid salt is selected from the group consisting of: O O O 9 3 181O O P - O O DSPS (L-isomer) – – O O O 9 3 P - DPPS (L-isomer) – – . In some embodiments, the anionic phospholipid salt is DSPS (L-isomer) sodium salt. In some embodiments, the anionic phospholipid salt is DSPS (L-isomer) ammonium salt. In some embodiments, the anionic phospholipid salt is DPPS (L-isomer) sodium salt. In some embodiments, the anionic phospholipid salt is DPPS (L-isomer) ammonium salt. In some embodiments the targeting lipid is a sodium or ammonium salt of dipalmitoylphosphatidyl-L- serine (DPPS) or distearoylphosphatidyl-L-serine (DSPS). In some embodiments the targeting 83 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 lipid is a sodium or ammonium salt of dipalmitoylphosphatidyl-L-serine (DPPS) or distearoylphosphatidyl-L-serine (DSPS). In some embodiments, a LNP composition can comprise an anionic phospholipid selected from the group consisting of: O O 9 3 OH 18P 161O O O OH DSPG 14 10 - In some embodiments, the salt form of phosphatidylserine is highly soluble in ethanol. In some embodiments it is soluble at greater than 0.5 mg / ml, greater than 1 mg / mL, greater than 5 84 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 mg / mL, greater than 10 mg / mL, or greater than 20 mg / mL. In some embodiments, the salt is an ammonium salt. In some embodiments, the salt is ammonium itself, an alkylammonium, a dialkylammonium, or a trialkylammonium salt. In some embodiments, the amine is chosen from ammonia, dimethylamine, diethylamine, triethylamine, trimethylamine, 2- (dimethyamino)ethanol, diethanolamine, 2-(diethyamino)ethanol, ethanolamine, ethylenediamine, N-methyl-glucamine, imidazole, histidine, lysine, arginine, 4-(2-hydroxyethyl)-morpholine, piperazine, 1-(2-hydroxyethyl)-pyrrolidine, triethanolamine, and tromethamine (tris(hydroxymethyl)aminomethane), In some embodiments, this targeting lipid is an ammonium salt of DPPS. Anionic phospholipids, separate from phosphatidyl-L-serine, were also considered as targeting lipids for LNPs. These include phosphatidylglycerol (PG), phosphatidic acid (PA), N- glutaryl-phosphatidylethanolamine (N-Glu-PE), N-succinyl-phosphatidylethanolamine (N-Suc- PE), and cardiolipin. In some embodiments, a LNP comprises anionic phospholipids, separate from phosphatidyl-L-serine, useful as targeting lipids for LNPs. In some embodiments, a LNP comprises anionic phospholipids selected from the group consisting of: phosphatidylglycerol (PG), phosphatidic acid (PA), N-glutaryl-phosphatidylethanolamine (N-Glu-PE), N-succinyl- phosphatidylethanolamine (N-Suc-PE), and cardiolipin. Distearoylphosphatidylglycerol (DSPG), dipalmitoyphosphatidylglycerol (DPPG), N-succinyl-distearoylphosphatidylethanolamine (N- Suc-DSPE), N-glutaryl-distearoylphosphatidylethanolamine (N-glu-DSPE), distearoylphosphatidic acid (DSPA), and cardiolipin are also provided as anionic phospholipids. In some embodiments, lipid nanoparticle (LNP) compositions comprising an ionizable cationic lipid compositions are provided. In some embodiments, lipid nanoparticle (LNP) compositions comprising an ionizable cationic lipid are provided. In some embodiments, the LNP composition comprises a mRNA nucleic acid. In some embodiments, a lipid nanoparticle (LNP) composition further comprises the PS lipid in a total amount of 2.5-10 mol% of the total lipid in the composition of the LNP. In some embodiments, a lipid nanoparticle (LNP) composition further comprises a PS lipid selected from the group consisting of: DSPS (L-isomer) and DPPS. In some embodiments, a lipid nanoparticle (LNP) composition comprises a conjugated lipid in a total amount of 0.5-2.0 mol% of the total lipid content of the LNP composition. In some embodiments, a lipid nanoparticle (LNP) composition comprises the conjugated lipid in a total 85 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 amount of less than 2 mol% of the total lipid content of the LNP composition, and the conjugated lipid is PEG-DMG. In some embodiments, a lipid nanoparticle (LNP) composition comprises a nucleic acid; an ionizable lipid disclosed herein; a sterol; one or more phospholipids comprising a phosphatidylserine (PS) lipid; and optionally further comprising a conjugated lipid. In some embodiments, a lipid nanoparticle (LNP) composition comprises a mRNA nucleic acid; an ionizable lipid disclosed herein; cholesterol; one or more phospholipids selected from the group consisting of: SM, DSPC, HSPC, DPPC and DOPC; and a PS lipid selected from the group consisting of: DPPS and DSPS; and optionally further comprising a conjugated lipid comprising PEG. In some embodiments, a nucleic acid lipid nanoparticle (LNP) composition comprises: a nucleic acid; an ionizable cationic lipid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; a sterol in a total amount of 25-45 mol% of the total lipid content of the LNP composition; and one or more phospholipids in a total amount of phospholipids of 5-25 mol% of the total lipid content of the LNP composition, and comprising a phosphatidylserine (PS) in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition; and optionally further comprising a conjugated lipid in a total amount of 0.5 – 2.5 mol% of the total lipid content of the LNP composition. In some embodiments, the LNP composition further comprises an anionic lipid selected from the group consisting of: DSPS (L-isomer), DPPS (L-isomer), DMPS (L-isomer), DOPS (L- isomer), and DSPS (D-isomer). Aspects of the disclosure relate to a lipid nanoparticle (LNP) composition comprising an ionizable lipid having the chemical structure: , or 4; 86 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 R2and R3are each independently (C1-C4) alkyl optionally substituted with hydroxyl; and n is an integer equal to 2, 3 or 4. In some embodiments, n is 2 or 3. In some embodiments, a is 0. In some embodiments, b is 1, 2 or 3. In some embodiments, a is 1. In some embodiments, b is 1, 2 or 3. In some embodiments, R2and R3are each methyl. In some embodiments, R1 is , In some embodiments, n is 3. In some embodiments, the comprises a nucleic acid; the ionizable lipid described herein, a sterol; one or more phospholipids comprising a phosphatidylserine (PS) lipid; and optionally a conjugated lipid. In some embodiments, the nucleic acid is mRNA. In some embodiments, the sterol is cholesterol. In other embodiments, the sterol is beta- sitosterol. In some embodiments, the one or more phospholipids consist of: one or more phospholipids selected from the group consisting of: SM, DSPC, HSPC, DPPC and DOPC; and a PS lipid selected from the group consisting of: DPPS, and DSPS. In some embodiments, the one or more phospholipids consist of: DSPC; and one or more PS lipids selected from the group consisting of (L-Serine) DPPS and (L-Serine) DSPS. In some embodiments, the composition comprises the PS lipid in a total amount of 2.5-10 mol% of the total lipid in the composition. In some embodiments, the conjugated lipid comprises PEG. Aspects of the disclosure relate to a nucleic acid lipid nanoparticle (LNP) composition comprising: a nucleic acid; an ionizable cationic lipid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; a sterol in a total amount of 25-45 mol% of the total lipid content of the LNP composition; and one or more phospholipids in a total amount of phospholipids of 5-25 mol% of the total lipid content of the LNP composition, and comprising a phosphatidylserine (PS) in a total amount of 2.5-10 mol% of the total lipid content of the LNP 87 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 composition; and optionally a conjugated lipid in a total amount of 0.5 – 2.5 mol% of the total lipid content of the LNP composition. In some embodiments, the nucleic acid is mRNA. In some embodiments, the sterol is cholesterol. In other embodiments, the sterol is beta- sitosterol. In some embodiments, the one or more phospholipids consist of: DSPC and a L-serine PS. In some embodiments, the composition comprises the PS in a total amount of 2.5-7.5 mol% of the total lipid in the composition. In some embodiments, the conjugated lipid comprises PEG. In some embodiments, conjugated lipid is PEG-DMG. In some embodiments, the LNP comprises the conjugated lipid in a total amount of 0.5-2.0 mol% of the total lipid content of the LNP composition. In some embodiments, the conjugated lipid in a total amount of less than 2 mol% of the total lipid content of the LNP composition. In some embodiments, the nucleic acid is a mRNA, the ionizable cationic lipid in a total amount of 45-55 mol% of the total lipid content of the LNP composition; a sterol is cholesterol in a total amount of 35-45 mol% of the total lipid content of the LNP composition; the total amount of phospholipid of 7-15 mol% of the total lipid content of the LNP composition; the one or more phospholipids consist of DSPC and the PS lipid is one or more lipids selected from the group consisting of the L-serine configuration of DPPS and DSPS; and the total amount of the PS lipid is about 5 mol% of the total lipid content of the LNP composition. In some embodiments, the composition comprises the PS lipid in a total amount selected from 1.25 mol%, 2.5 mol%, 5 mol%, 7.5 mol%, and 10 mol% of the total lipid content of the LNP composition. Aspects of the disclosure relate to a nucleic acid lipid nanoparticle (LNP) composition comprising: a nucleic acid, wherein the nucleic acid is mRNA; an ionizable cationic lipid, the ionizable cationic lipid in a total amount of 45-55 mol% of the total lipid content of the LNP composition; a sterol, wherein the sterol is cholesterol in a total amount of 35-45 mol% of the total lipid content of the LNP composition; one or more phospholipids, wherein the one or more phospholipids in a total amount of phospholipids of 10 mol% of the total lipid content of the LNP composition, and comprising a phosphatidylserine (PS) in a total amount of 3-9 mol% of the total 88 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 lipid content of the LNP composition; and a conjugated lipid, the conjugated lipid in a total amount of 0.5 – 2.0 mol% of the total lipid content of the LNP composition. In some embodiments, the one or more phospholipid is selected from the group consisting of: DSPS (L-isomer), DPPS (L-isomer), DMPS (L-isomer), DOPS (L-isomer), and DSPS (D-isomer). In some embodiments, the conjugated lipid is PEG-DMG; and the PS lipid is selected from the group consisting of: DSPS (L-isomer) and DPPS. In some embodiments, the ionizable cationic lipid is one or more compounds selected from the group consisting of: KC3-OA, KC3-PA, KC3-C17 (8:1), and KC3-C15 (C8:1). In some embodiments, the ionizable cationic lipid is KC3-PA. In some embodiments, the ionizable cationic lipid is KC3-OA. In some embodiments, the ionizable cationic lipid is KC3-C17 (C8:1). In some embodiments, the LNP comprises a nucleic acid; an ionizable cationic lipid in a total amount of 50 mol% of the total lipid content of the LNP composition; cholesterol in a total amount of 38.5 mol% of the total lipid content of the LNP composition; one or more phospholipids in a total amount of 7-15 mol% of the total lipid content of the LNP composition, and comprising a phosphatidylserine (PS) lipid in a total amount of 3-9 mol% of the total lipid content of the LNP composition; and a PEG-containing lipid in a total amount of 0.5 –2.0 mol% of the total lipid content of the LNP composition. In some embodiments, the phospholipids consist of one or more phospholipids selected from the group consisting of: DSPC, DOPC, DPPC, HSPC, and SM. In some embodiments, the PS lipid is one or more L-serine lipids selected from the group consisting of DPPS and DSPS. In some embodiments, the one or more phospholipids comprise at least two (L-Serine) PS lipids having mismatched acyl chain lengths. In some embodiments, the one or more phospholipids comprise at least two phospholipids having mismatched acyl chain lengths. In some embodiments, the one or more phospholipids comprises distearoylphosphatidylcholine (DSPC), hydrogenated soy phosphatidylcholine (HSPC), dipalmitoylphosphatidyl-L-serine ((L-serine)DPPS) or a combination thereof. In some embodiments, the phospholipids are DSPC and DPPS. In some embodiments, the DSPC and DPPS are each present in the LNP at a total amount of 5 mol% each, based on the total lipid content of the LNP composition. 89 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 In some embodiments, the ionizable cationic lipid is KC4-OA. Aspects of the disclosure relate to a nucleic acid lipid nanoparticle (LNP) composition comprising an ionizable cationic lipid selected from one or more of the following a mixture of 3- (S)-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N-dimethylpropan-1-amine (KC3-OA(S)) and 3-(R)-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N-dimethylpropan-1-amine (KC3- OA(R)) enantiomer; and 4-(S)-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N- dimethylbutan-1-amine (KC4-OA(S)) and 4-(R)-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4- yl)-N,N-dimethylbutan-1-amine (KC4-OA(R)). In some embodiments, the ionizable cationic lipid is a racemic mixture of 3-rac-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N- dimethylpropan-1-amine (KC3-OA racemate). In other embodiments, the ionizable cationic lipid is a racemic mixture of 4-rac-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N- dimethylbutan-1-amine (KC4-OA racemate). In other embodiments, the ionizable cationic lipid is 3-(R)-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N-dimethylpropan-1-amine (KC3- OA(R)). Yet, in other embodiments, the ionizable cationic lipid is 4-(R)-2,2-di((Z)-octadec-9-en- 1-yl)-1,3-dioxolan-4-yl)-N,N-dimethylbutan-1-amine (KC4-OA(R)). In some embodiments, the ionizable cationic lipid is KC3-OA racemate of KC3-OA(S) and KC3-OA(R). In some embodiments, the ionizable cationic lipid is KC3-OA (R) enantiomer. In some embodiments, the ionizable cationic lipid is a KC3-OA enantiomer purified from AKG-KC3- OA racemate of KC3-OA(S) and KC3-OA(R). In some embodiments, the ionizable cationic lipid is KC3-OA mixed enantiomers of KC3-OA(S) and KC3-OA(R), or KC4-OA mixed enantiomers of KC4-OA(S) and KC4-OA(R). Aspects of the disclosure relate to a nucleic acid lipid nanoparticle (LNP) composition comprising: a nucleic acid, ionizable cationic lipid KC3-PA or KC3-OA, and a (L-Serine) PS lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition. In some embodiments, the nucleic acid is mRNA, the PS lipid is (L-Serine) DSPS, (L-Serine) DPPS, or a mixture thereof, and the LNP composition further comprises cholesterol and a second phospholipid selected from the group consisting of: DSPC, DPPC, HSPC, and SM. In some embodiments, the LNP composition further comprises 0.5-2.0 mol% PEG-DMG or PEG-DSG, based on the total lipid content in the LNP composition. In some embodiments, the ionizable cationic lipid is KC3-PA. In some embodiments, the ionizable cationic lipid KC3-OA. 90 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 Aspects of the disclosure relate to a nucleic acid lipid nanoparticle (LNP) composition comprising: a nucleic acid, a KC3-C17 (C8:1) ionizable cationic lipid; and a (L-Serine) PS lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition. In some embodiments, the LNP composition has a N / P ratio 4 to 7. In some embodiments, the composition has a N / P ratio of 5 to 6. In some embodiments, the composition has a N / P ratio of 5.3. Aspects of the disclosure relate to a nucleic acid lipid nanoparticle (LNP) composition comprising: a nucleic acid, ionizable cationic lipid KC3-PA, and a (L-Serine) PS lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition. In some embodiments, the nucleic acid is mRNA, the PS lipid is (L-Serine) DSPS, (L-Serine) DPPS, or a mixture thereof, and the LNP composition further comprises cholesterol and a second phospholipid selected from the group consisting of: DSPC, DOPC, DPPC, HSPC, and SM. In some embodiments, the LNP composition further comprises 0.5-2.0 mol% PEG-DMG or PEG-DSG, based on the total lipid content in the LNP composition. Aspects of the disclosure relate to a nucleic acid lipid nanoparticle (LNP) composition comprising: a nucleic acid, an ionizable cationic lipid selected from KC3-C17 (C8:1); and a (L- Serine) PS lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition. In some embodiments, the N / P ratio is 4 to 7. In some embodiments, the N / P ratio is 5 to 6. In some embodiments, the N / P ratio is 3. In some embodiments, the N / P ratio is 7. In some embodiments, the nucleic acid is mRNA encoding SARS-CoV-2 spike protein. Aspects of the disclosure relate to a nucleic acid lipid nanoparticle (LNP) vaccine composition comprising: a mRNA nucleic acid with a N / P ratio of 4 to 7; an KC3-PA ionizable cationic lipid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; cholesterol in a total amount of 25-40 mol% of the total lipid content of the LNP composition; a (L-Serine) PS lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition; DSPC phospholipid in a total amount of 5-25 mol% of the total lipid content of the LNP composition; and PEG-DMG in a total amount of 0-2.5 mol% of the total lipid content of the LNP composition. Aspects of the disclosure relate to a nucleic acid lipid nanoparticle (LNP) vaccine composition comprising: a mRNA nucleic acid with a N / P ratio of 3 to 8; a KC3-C17 (C8:1) ionizable cationic lipid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; cholesterol in a total amount of 25-40 mol% of the total lipid content of the LNP composition; a (L-Serine) 91 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 PS lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition; DSPC phospholipid in a total amount of 5-25 mol% of the total lipid content of the LNP composition; and PEG-DMG in a total amount of 0-2.5 mol% of the total lipid content of the LNP composition. Aspects of the disclosure relate to a nucleic acid lipid nanoparticle (LNP) vaccine composition comprising: a mRNA nucleic acid with a N / P ratio of 4 to 7; a KC3-C15 (C8:1) ionizable cationic lipid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; cholesterol in a total amount of 25-40 mol% of the total lipid content of the LNP composition; a (L-Serine) PS lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition; DSPC phospholipid in a total amount of 5-25 mol% of the total lipid content of the LNP composition; and PEG-DMG in a total amount of 0-2.5 mol% of the total lipid content of the LNP composition. Aspects of the disclosure relate to a nucleic acid lipid nanoparticle (LNP) vaccine composition comprising: a mRNA nucleic acid with a N / P ratio of 3 to 8; a KC3-C18 ionizable cationic lipid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; cholesterol in a total amount of 25-40 mol% of the total lipid content of the LNP composition; a (L-Serine) PS lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition; DSPC phospholipid in a total amount of 5-25 mol% of the total lipid content of the LNP composition; and PEG-DMG in a total amount of 0-2.5 mol% of the total lipid content of the LNP composition. In some embodiments, the nucleic acid is a mRNA of SEQ ID NO: 2. Aspects of the disclosure relate to the use of a (L-Serine) PS lipid in combination with an ionizable cationic lipid described herein in the LNP for targeting of the LNP to dendritic cells. In some embodiments, the LNP comprises mRNA. In some embodiments, the LNP further comprises cholesterol. In some embodiments, the total amount of (L-Serine) PS lipid in the LNP is 2.5-10 mol% of the total lipid content of the LNP composition. In some embodiments, the LNP further comprises one or more additional phospholipids including DSPC. In some embodiments, the LNP further comprises a conjugated lipid. In some embodiments, the LNP comprises: a mRNA nucleic acid with a N / P ratio of 3 to 8; a KC3-PA or KC3-C17 (C8:1) ionizable cationic lipid (ICL), in a total amount of 40-65 mol% of the total lipid content of the LNP composition; cholesterol in a total amount of 25-40 mol% of the total lipid content of the LNP composition; a (L-Serine) PS lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition; DSPC phospholipid in a total amount of 5-25 mol% of the total lipid content of the LNP composition; and a conjugated lipid in a total amount of 0-2.5 mol% of the total lipid content of the LNP 92 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 composition. In some embodiments, the ICL is KC3-PA. In some embodiments, the ICL is KC3- C17 (C8:1). Some aspects of the disclosure relate to a lipid nanoparticle (LNP) composition comprising an ionizable lipid having the chemical structure: , or 4; R2and R3are each independently methyl; and n is an integer equal to 2 or 3. In some embodiments, a is 0. In some embodiments, b is 1. In some embodiments, b is 3. In some embodiments, a is 1. In some embodiments, b is 1. In some embodiments, b is 3. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, the composition comprises an anionic lipid selected from the group consisting of: phosphatidylglycerol (PG), phosphatidic acid (PA), N-glutaryl- phosphatidylethanolamine (N-Glu-PE), N-succinyl-phosphatidylethanolamine (N-Suc-PE), and cardiolipin. Distearoylphosphatidylglycerol (DSPG), dipalmitoyphosphatidylglycerol (DPPG), N-succinyl-distearoylphosphatidylethanolamine (N-Suc-DSPE), N-glutaryl- distearoylphosphatidylethanolamine (N-glu-DSPE), distearoylphosphatidic acid (DSPA), and cardiolipin. In some embodiments, the composition comprises an anionic targeting phospholipid other than phosphatidyl-L-serine. In some embodiments, the composition comprises an anionic phospholipid selected from the group consisting of: DSPG and DPPG. In some embodiments, the composition comprises an anionic phospholipid selected from the group consisting of: N-Glu-DSPE and N-Suc-DSPE. 93 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 In some embodiments, the composition comprises a DSPA anionic phospholipid. In some embodiments, the composition comprises a Cardiolipin anionic phospholipid. In some embodiments, the ionizable lipid has the chemical structure: . lipid has the chemical structure: . to a sodium or ammonia salt of a composition of an anionic phospholipid of Formula (V-A-1), having the chemical structure: wherein X+is an ammonium cation or a sodium (Na+) cation; and a is 14, 15 or 16. In some embodiments, a is 14 or 16. In some embodiments, X+is ammonium cation (NH4+). In some embodiments, X+is sodium cation (Na+) In some embodiments, X is an ammonium cation selected from the group consisting of: ammonium (NH4+), an alkylammonium, a dialkylammonium, and a trialkylammonium salt. In some embodiments, X is X is an ammonium cation selected from the group consisting of: ammonium, dimethylamine, diethylamine, triethylamine, trimethylamine, 2- (dimethyamino)ethanol, diethanolamine, 2-(diethyamino)ethanol, ethanolamine, ethylenediamine, N-methyl-glucamine, imidazole, histidine, lysine, arginine, 4-(2-hydroxyethyl)-morpholine, 94 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 piperazine, 1-(2-hydroxyethyl)-pyrrolidine, triethanolamine, and tromethamine (tris(hydroxymethyl)aminomethane). In some embodiments, the anionic phospholipid of Formula (V-A-1) is a sodium salt of distearoylphosphatidyl-L-serine (DSPS L-isomer). In some embodiments, the anionic phospholipid of Formula (V-A-1) is an ammonium salt of distearoylphosphatidyl-L-serine (DSPS L-isomer). In some embodiments, the anionic phospholipid of Formula (V-A-1) is a sodium salt of DPPS (L-isomer). In some embodiments, the anionic phospholipid of Formula (V-A-1) is an ammonium salt of DPPS (L-isomer). Some embodiments relate to the use of the salt form composition in the preparation of a liposomal nanoparticle (LNP) composition. In some embodiments, the use is in combination with one or more of the following LNP components during the preparation of the LNP composition: a mRNA nucleic acid; an ionizable cationic lipid (ICL); cholesterol; a (L-Serine) PS lipid; one or more phospholipids; and a conjugated lipid. In some embodiments, the use comprises the step of combining the ammonium or salt form of a compound of Formula (V-A-1) with one or more of the following LNP components during the preparation of the LNP composition: a mRNA nucleic acid; an ionizable cationic lipid (ICL); cholesterol; a (L- Serine) PS lipid; one or more phospholipids; and a conjugated lipid. In some embodiments, the LNP is a nucleic acid lipid nanoparticle vaccine composition comprising: a mRNA nucleic acid with a N / P ratio of 4 to 7; an ionizable cationic lipid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; cholesterol in a total amount of 25-40 mol% of the total lipid content of the LNP composition; a (L-Serine) PS lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition; DSPC phospholipid in a total amount of 5-25 mol% of the total lipid content of the LNP composition; and PEG-DMG in a total amount of 0-2.5 mol% of the total lipid content of the LNP composition. In some embodiments, the nucleic acid is mRNA of SEQ ID NO: 2. In some embodiments, the LNP composition comprises the ionizable cationic lipid in a total amount of 46-65 mol% of the total lipid content of the LNP composition. In some embodiments, the LNP composition comprises the PS in a total amount of about 5 mol% of the total lipid in the composition. In some embodiments, the LNP composition comprises the conjugated lipid in a total amount of about 1.5 mol% of the total lipid content of the LNP composition. In some embodiments, the conjugated lipid is PEG-DMG; and the PS lipid is selected from the group consisting of: DSPS (L-isomer) and DPPS. 95 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 In some embodiments, the ionizable cationic lipid is one or more compounds selected from the group consisting of: KC3-OA, KC3-PA, KC3-C17 (C8:1), and KC3-C15 (C8:1). In some embodiments, the ionizable cationic lipid is KC3-PA. In some embodiments, the ionizable cationic lipid is KC3-OA. In some embodiments, the ionizable cationic lipid is KC3-C17 (C8:1). Some embodiments relate to the use of a (L-Serine) PS lipid in combination with an ionizable cationic lipid described herein in the LNP for targeting of the LNP to dendritic cells. In some embodiments, the LNP comprises mRNA. In some embodiments, the LNP further comprises cholesterol. In some embodiments, the total amount of (L-Serine) PS lipid in the LNP is 2.5-10 mol% of the total lipid content of the LNP composition. In some embodiments, the LNP further comprises one or more additional phospholipids including DSPC. In some embodiments, the LNP further comprises a conjugated lipid. In some embodiments, the LNP comprises: a mRNA nucleic acid with a N / P ratio of 3 to 8; a KC3-PA or KC3-C17 (C8:1) ionizable cationic lipid (ICL), in a total amount of 40-65 mol% of the total lipid content of the LNP composition; cholesterol in a total amount of 25-40 mol% of the total lipid content of the LNP composition; a (L-Serine) PS lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition; DSPC phospholipid in a total amount of 5-25 mol% of the total lipid content of the LNP composition; and a conjugated lipid in a total amount of 0-2.5 mol% of the total lipid content of the LNP composition. In some embodiments, the ICL is KC3-PA. In some embodiments, the ICL is KC3-C17 (C8:1). In some embodiments, the composition comprises an anionic phospholipid selected from the group consisting of: DSPG and DPPG, in a total amount of 2.5-7.5% of the total lipid content of the LNP composition. In some embodiments, the composition comprises DSPG anionic phospholipid in a total amount of 2.5-7.5% of the total lipid content of the LNP composition. In some embodiments, the composition comprises DPPG anionic phospholipid in a total amount of 2.5-7.5% of the total lipid content of the LNP composition. In some embodiments, the LNP further comprises one or more additional phospholipids including DSPC. Aspects of the disclosure relate to a nucleic acid lipid nanoparticle (LNP) composition comprising: a nucleic acid; a KC3 ionizable cationic lipid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; cholesterol in a total amount of 23.5 - 43.5 mol% of the total lipid content of the LNP composition; a (L-Serine) PS lipid in a total amount of 2.5-10 96 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 mol% of the total lipid content of the LNP composition; DSPC or HSPC phospholipid in a total amount of 5-25 mol% of the total lipid content of the LNP composition; and a PEG-containing conjugated lipid in a total amount of 0.5 mol% to 2.5 mol% of the total lipid content of the LNP composition. In some embodiments, the nucleic acid is mRNA. In some embodiments, the N / P ratio is 3 to 8. In some embodiments, the KC3 ionizable cationic lipid is selected from the group consisting of: KC3-OA, KC3-PA, KC3-C17 (8:1), and KC3-C15 (C8:1). In some embodiments, the KC3 ionizable cationic lipid is KC3-OA. In some embodiments, the KC3 ionizable cationic lipid is KC3-PA. In some embodiments, the KC3 ionizable cationic lipid is KC3-C17(C8:1). In some embodiments, the KC3 ionizable cationic lipid is KC3-C15(C8:1). In some embodiments, the conjugated lipid is PEG-DMG or PEG-DSG. In some embodiments, the composition comprises the PEG-containing conjugated lipid in a total amount of 0.5 –2.0 mol% of the total lipid content of the LNP composition. In some embodiments, the composition comprises the KC3 ionizable cationic lipid in a total amount of 48 mol% of the total lipid content of the LNP composition. In some embodiments, the composition comprises DSPC and DSPS in a total amount of 10 mol% of the total lipid content of the LNP composition. In some embodiments, the composition comprises 5 % DSPC or HSPC in a total amount of 5 mol% of the total lipid content of the LNP composition. In some embodiments, the composition comprises PEG-DMG in a total of 1.5 mol % of the total lipid content of the LNP composition. In some embodiments, the composition comprises cholesterol in a total amount of 40.5 mol % cholesterol of the total lipid content of the LNP composition. In some embodiments, the composition comprises the DSPC phospholipid in a total amount of 10 mol% of the total lipid content of the LNP composition. In some embodiments, the PEG-containing conjugated lipid is PEG2000-DMG. In some embodiments, the composition comprises the cholesterol in a total amount of 23.5 mol% of the total lipid content of the LNP composition. In some embodiments, the composition comprises the cholesterol in a total amount of 33.5 mol% of the total lipid content of the LNP composition. In some embodiments, the composition comprises the cholesterol in a total amount 97 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 of 38.5 mol% of the total lipid content of the LNP composition. In some embodiments, the composition comprises the cholesterol in a total amount of 40.5 mol% of the total lipid content of the LNP composition. In some embodiments, the composition comprises the cholesterol in a total amount of 42.7 mol% of the total lipid content of the LNP composition. b In some embodiments, the composition comprises the cholesterol in a total amount of 43.5 mol% of the total lipid content of the LNP composition. In some embodiments, the composition comprises the cholesterol in a total amount of 33.5-43.5 mol% of the total lipid content of the LNP composition. In some embodiments, the composition comprises the KC3 ionizable cationic lipid in a total amount of 45- 55 mol% of the total lipid content of the LNP composition. Aspects of the disclosure relate to a nucleic acid lipid nanoparticle (LNP) composition comprising: a mRNA nucleic acid; a KC3 ionizable cationic lipid selected from the group consisting of: KC3-OA, KC3-PA, KC3-C17 (8:1), and KC3-C15 (C8:1), in a total amount of 45- 55 mol% of the total lipid content of the LNP composition; cholesterol in a total amount of 33.5- 43.5 mol% of the total lipid content of the LNP composition; a (L-Serine) DPPS lipid in a total amount of 5 mol% of the total lipid content of the LNP composition; DSPC or HSPC phospholipid in a total amount of 5 mol% of the total lipid content of the LNP composition; and a PEG-DMG conjugated lipid in a total amount of 1.5 mol% of the total lipid content of the LNP composition. Aspects of the disclosure relate to a lipid nanoparticle (LNP) composition comprising a KC3 ionizable cationic lipid, a (L-Serine) PS lipid, cholesterol, one or more phospholipids comprising at least one anionic phospholipid, and a conjugated lipid, wherein the LNP is obtained by a process comprising the step of dissolving a sodium or ammonium salt of the anionic phospholipid. In some embodiments, the composition comprises a nucleic acid. In some embodiments, the nucleic acid is mRNA. In some embodiments, the composition is a vaccine. In some embodiments, the total amount of phospholipids in the composition is 5-25 mol% of the total lipid content of the LNP composition, and the total amount of the phosphatidylserine (PS) is 2.5-10 mol% of the total lipid content of the LNP composition; and the total amount of the conjugated lipid in the composition is a total amount of 0.5 – 2.5 mol% of the total lipid content of the LNP composition. 98 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 In some embodiments, the composition comprises 48 mol% of the KC3 ionizable cationic lipid, 40.5 mol% cholesterol, and 5 mol% (L-Serine) DPPS lipid, wherein each mol% refers to the mol% of the total lipid content of the LNP composition. In some embodiments, the composition comprises 48 mol% of the KC3 ionizable cationic lipid, 38.5 mol% cholesterol, and 5 mol% (L-Serine) DPPS lipid, wherein each mol% refers to the mol% of the total lipid content of the LNP composition. In some embodiments, the composition comprises 46-54 mol% of the KC3 ionizable cationic lipid, and 5 mol% (L-Serine) DPPS lipid, wherein each mol% referes to the mol% of the total lipid content of the LNP composition. In some embodiments, the composition comprises 45 mol% of the KC3 ionizable cationic lipid, 42.7 mol% cholesterol, and 5 mol% (L-Serine) DPPS lipid, wherein each mol% refers to the mol% of the total lipid content of the LNP composition. In some embodiments, the composition comprises 50 mol% of the KC3 ionizable cationic lipid, 38.5 mol% cholesterol, 5 mol% (L-Serine) DPPS lipid, and a total of 10 mol% phospholipid concentration; wherein each mol% refers to the mol% of the total lipid content of the LNP composition. In some embodiments, the composition comprises 48 mol% of the KC3 ionizable cationic lipid, 40.5 mol% cholesterol, 5 mol% (L-Serine) DPPS lipid, anda total of 10 mol% phospholipid concentration; wherein each mol% refers to the mol% of the total lipid content of the LNP composition. In some embodiments, the composition comprises 48 mol% of the KC3 ionizable cationic lipid, 40.5 mol% cholesterol, 5 mol% (L-Serine) DPPS lipid, 5 mol% DSPC or DPPC; anda total of 10 mol% phospholipid concentration; wherein each mol% refers to the mol% of the total lipid content of the LNP composition. In some embodiments, the composition comprises 46.5 mol% of the KC3 ionizable cationic lipid, 42 mol% cholesterol, 5 mol% (L-Serine) DPPS lipid, wherein each mol% refers to the mol% of the total lipid content of the LNP composition. In some embodiments, the composition further comprises a total of 5 mol% DSPC or HSPC of the total lipid content of the LNP composition. In some embodiments, the composition further comprises a total of 1.5 mol% PEG-DMG of the total lipid content of the LNP composition. 99 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 In some embodiments, the composition comprises a total of 10 mol% of DSPC / DPPC phospholipid of the total lipid content of the LNP composition. Aspects of the disclosure relate to a phosphatidylserine salt selected from the group consisting of DSPS sodium, DPPS sodium, DSPS ammonium and DPPS ammonium. Aspects of the disclosure relate to the use of a DSPS-Na salt or a DPPS-NH4+salt in the preparation of a LNP comprising a (L-Serine) PS lipid, a sterol, a conjugated lipid, a phospholipid for targeting the LNP to dendritic cells. Aspects of the disclosure relate to a solution comprising ethanol and DSPS or DPPS, the solution obtained by a process comprising the step of dissolving a phosphatidylserine salt in ethanol, wherein the phosphatidylserine salt is selected from the group consisting of DSPS sodium, DPPS sodium, DSPS ammonium and DPPS ammonium. Embodiments: Non-limiting embodiments are described below each of which is considered to be within the present disclosure. Embodiment 1: A lipid nanoparticle (LNP) composition comprising an ionizable lipid having the chemical structure: , 1, 2, 3 or 4; R2and R3are each independently (C1-C4) alkyl optionally substituted with hydroxyl; and n is an integer equal to 2, 3 or 4. Embodiment 2: The composition of embodiment 1, wherein n is 2 or 3. Embodiment 3: The composition of any one of embodiments 1 - 2, wherein a is 0. Embodiment 4: The composition of any one of embodiments 1 - 3, wherein b is 1, 2 or 3. Embodiment 5: The composition of any one of embodiments 1 - 2, wherein a is 1. 100 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 Embodiment 6: The composition of any one of embodiments 1 - 3, wherein b is 1, 2 or 3. Embodiment 7: The composition of any one of embodiments 1 - 6, wherein R2 and R3 are each methyl. Embodiment 8: The composition of embodiment 1, wherein R1is , R2 and R3 are each methyl; and n is 2 or 3. Embodiment 9: The composition of any one of embodiments 1-8, wherein n is 3. Embodiment 10: A composition of any one of embodiments 1-9, comprising: a. a nucleic acid; b. the ionizable lipid of any one of embodiments 1-9; c. a sterol; d. one or more phospholipids comprising a phosphatidylserine (PS) lipid; and e. optionally further comprising a conjugated lipid. Embodiment 11: The composition of embodiment 10, wherein the nucleic acid is mRNA. Embodiment 12: The composition of embodiment 11, wherein the sterol is cholesterol. Embodiment 13: The composition of embodiment 12, wherein the one or more phospholipids consist of: a. one or more phospholipids selected from the group consisting of: SM, DSPC, HSPC, DPPC and DOPC; and b. a PS lipid selected from the group consisting of: DPPS, and DSPS . Embodiment 14: The composition of embodiment 13, wherein the one or more phospholipids consist of: a. DSPC; and b. one or more PS lipids selected from the group consisting of (L-Serine) DPPS and (L- Serine) DSPS. Embodiment 15: The composition of embodiment 13, wherein the composition comprises the PS lipid in a total amount of 2.5-10 mol% of the total lipid in the composition. 101 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 Embodiment 16: The composition of any one of embodiments 10-15, wherein the conjugated lipid comprises PEG. Embodiment 17: A nucleic acid lipid nanoparticle (LNP) composition comprising: a. a nucleic acid; b. an ionizable cationic lipid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; c. a sterol in a total amount of 25-45 mol% of the total lipid content of the LNP composition; and d. one or more phospholipids in a total amount of phospholipids of 5-25 mol% of the total lipid content of the LNP composition, and comprising a phosphatidylserine (PS) in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition; and e. optionally further comprising a conjugated lipid in a total amount of 0.5 – 2.5 mol% of the total lipid content of the LNP composition. Embodiment 18: The composition of embodiment 17, wherein the nucleic acid is mRNA. Embodiment 19: The composition of embodiment 18, wherein the sterol is cholesterol. Embodiment 20: The composition of embodiment 19, wherein the one or more phospholipids consists of: DSPC and a L-serine PS. Embodiment 21: The composition of embodiment 20, wherein the composition comprises the PS in a total amount of 2.5-7.5 mol% of the total lipid in the composition. Embodiment 22: The composition of any one of embodiments 17-21, wherein the conjugated lipid comprises PEG. Embodiment 23: The composition of embodiment 22, wherein the conjugated lipid is PEG-DMG. Embodiment 24: The composition of embodiment 23, wherein the LNP comprises the conjugated lipid in a total amount of 0.5-2.0 mol% of the total lipid content of the LNP composition. Embodiment 25: The composition of embodiment 24, wherein the LNP comprises the conjugated lipid in a total amount of less than 2 mol% of the total lipid content of the LNP composition. Embodiment 26: The composition of embodiment 17, wherein: a. the nucleic acid is a mRNA, b. the ionizable cationic lipid in a total amount of 45-55 mol% of the total lipid content of the LNP composition; 102 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 c. a sterol is cholesterol in a total amount of 35-45 mol% of the total lipid content of the LNP composition; d. the total amount of phospholipid of 7-15 mol% of the total lipid content of the LNP composition; e. the one or more phospholipids consist of DSPC and the PS lipid is one or more lipids selected from the group consisting of the L-serine configuration of DPPS and DSPS; and f. the total amount of the PS lipid is about 5 mol% of the total lipid content of the LNP composition. Embodiment 27: The composition of embodiment 26, wherein the composition comprises the PS lipid in a total amount selected from 1.25 mol%, 2.5 mol%, 5 mol%, 7.5 mol%, and 10 mol% of the total lipid content of the LNP composition. Embodiment 28: A nucleic acid lipid nanoparticle (LNP) composition comprising: a. a nucleic acid, wherein the nucleic acid is mRNA; b. an ionizable cationic lipid, the ionizable cationic lipid in a total amount of 45-55 mol% of the total lipid content of the LNP composition; c. a sterol, wherein the sterol is cholesterol in a total amount of 35-45 mol% of the total lipid content of the LNP composition; d. one or more phospholipids, wherein the one or more phospholipids in a total amount of phospholipids of 10 mol% of the total lipid content of the LNP composition, and comprising a phosphatidylserine (PS) in a total amount of 3-9 mol% of the total lipid content of the LNP composition; and e. a conjugated lipid, the conjugated lipid in a total amount of 0.5 – 2.0 mol% of the total lipid content of the LNP composition. Embodiment 29: The composition of any one of embodiments 17- 28, wherein the one or more phospholipid is selected from the group consisting of: DSPS (L-isomer), DPPS (L-isomer), DMPS (L-isomer), DOPS (L-isomer), and DSPS (D-isomer). Embodiment 30: The composition of embodiment 29, wherein a. the conjugated lipid is PEG-DMG; and b. the PS lipid is selected from the group consisting of: DSPS (L-isomer) and DPPS. 103 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 Embodiment 31: The composition of any one of embodiments 17-28, wherein the ionizable cationic lipid is one or more compounds selected from the group consisting of: KC3-OA, KC3- PA, KC3-C17 (8:1), and KC3-C15 (C8:1). Embodiment 32: The composition of any one of embodiments 17-28, wherein the ionizable cationic lipid is KC3-PA. Embodiment 33: The composition of any one of embodiments 17-28, wherein the ionizable cationic lipid is KC3-OA. Embodiment 34: The composition of any one of embodiments 17-28, wherein the ionizable cationic lipid is KC3-C17 (C8:1). Embodiment 35: The nucleic acid lipid nanoparticle (LNP) composition of embodiment 17, comprising: a. a nucleic acid; b. an ionizable cationic lipid in a total amount of 50 mol% of the total lipid content of the LNP composition; c. cholesterol in a total amount of 38.5 mol% of the total lipid content of the LNP composition; d. one or more phospholipids in a total amount of 7-15 mol% of the total lipid content of the LNP composition, and comprising a phosphatidylserine (PS) lipid in a total amount of 3-9 mol% of the total lipid content of the LNP composition; and e. a PEG-containing lipid in a total amount of 0.5 –2.0 mol% of the total lipid content of the LNP composition. Embodiment 36: The composition of embodiment 34, wherein the phospholipids consist of one or more phospholipids selected from the group consisting of: DSPC, DOPC, DPPC, HSPC, and SM. Embodiment 37: The composition of embodiment 36, wherein the PS lipid is one or more L-serine lipids selected from the group consisting of DPPS and DSPS. Embodiment 38: The composition of any one of embodiments 17-28, wherein the one or more phospholipids comprise at least two (L-Serine) PS lipids having mismatched acyl chain lengths. Embodiment 39: The composition of embodiment 38, wherein the phospholipids are DSPC and DPPS. 104 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 Embodiment 40: The composition of embodiment 39, wherein the DSPC and DPPS are each present in the LNP at a total amount of 5 mol% each, based on the total lipid content of the LNP composition. Embodiment 41: A nucleic acid lipid nanoparticle (LNP) composition comprising: a nucleic acid, ionizable cationic lipid KC3-PA or KC3-OA, and a (L-Serine) PS lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition. Embodiment 42: The composition of embodiment 41, wherein the nucleic acid is mRNA, the PS lipid is (L-Serine) DSPS, (L-Serine) DPPS, or a mixture thereof, and the LNP composition further comprises cholesterol and a second phospholipid selected from the group consisting of: DSPC, DPPC, HSPC, and SM. Embodiment 43: The composition of embodiment 42, wherein the LNP composition further comprises 0.5-2.0 mol% PEG-DMG or PEG-DSG, based on the total lipid content in the LNP composition. Embodiment 44: The composition of any one of embodiments 41-43, wherein the ionizable cationic lipid is KC3-PA. Embodiment 45: The composition of any one of embodiments 41-43, wherein the ionizable cationic lipid KC3-OA. Embodiment 46: A nucleic acid lipid nanoparticle (LNP) composition comprising: a nucleic acid, a KC3-C17 (C8:1) ionizable cationic lipid; and a (L-Serine) PS lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition. Embodiment 47: The composition of any one of embodiments 41-46, wherein the LNP composition has a N / P ratio 4 to 7. Embodiment 48: The composition of embodiment 47, wherein the LNP composition has a N / P ratio of 5 to 6. Embodiment 49: The composition of embodiment 48, wherein the LNP composition has a N / P ratio of 5.3. Embodiment 50: A nucleic acid lipid nanoparticle (LNP) composition comprising: a nucleic acid, ionizable cationic lipid KC3-PA, and a (L-Serine) PS lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition. Embodiment 51: The composition of embodiment 50, wherein the nucleic acid is mRNA, the PS lipid is (L-Serine) DSPS, (L-Serine) DPPS, or a mixture thereof, and the LNP composition further 105 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 comprises cholesterol and a second phospholipid selected from the group consisting of: DSPC, DOPC, DPPC, HSPC, and SM. Embodiment 52: The composition of embodiment 51, wherein the LNP composition further comprises 0.5-2.0 mol% PEG-DMG or PEG-DSG, based on the total lipid content in the LNP composition. Embodiment 53: A nucleic acid lipid nanoparticle (LNP) composition comprising: a nucleic acid, an ionizable cationic lipid selected from KC3-C17 (C8:1); and a (L-Serine) PS lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition. Embodiment 54: The composition of any one of embodiments 17-21, 26-28, 35-44, or 53, wherein the N / P ratio is 4 to 7. Embodiment 55: The composition of embodiment 54, wherein the N / P ratio is 5 to 6. Embodiment 56: The composition of embodiment 55, wherein the N / P ratio is 3. Embodiment 57: The composition of embodiment 55, wherein the N / P ratio is 7. Embodiment 58: The composition of any one of embodiments 17-21, 26-28, 35-47, or 53, wherein the nucleic acid is mRNA encoding SARS-CoV-2 spike protein. Embodiment 59: A nucleic acid lipid nanoparticle (LNP) vaccine composition comprising: a. a mRNA nucleic acid with a N / P ratio of 4 to 7; b. an KC3-PA ionizable cationic lipid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; c. cholesterol in a total amount of 25-40 mol% of the total lipid content of the LNP composition; d. a (L-Serine) PS lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition; e. DSPC phospholipid in a total amount of 5-25 mol% of the total lipid content of the LNP composition; and f. PEG-DMG in a total amount of 0-2.5 mol% of the total lipid content of the LNP composition. Embodiment 60: A nucleic acid lipid nanoparticle (LNP) vaccine composition comprising: a. a mRNA nucleic acid with a N / P ratio of 3 to 8; b. a KC3-C17 (C8:1) ionizable cationic lipid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; 106 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 c. cholesterol in a total amount of 25-40 mol% of the total lipid content of the LNP composition; d. a (L-Serine) PS lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition; e. DSPC phospholipid in a total amount of 5-25 mol% of the total lipid content of the LNP composition; and f. PEG-DMG in a total amount of 0-2.5 mol% of the total lipid content of the LNP composition. Embodiment 61: A nucleic acid lipid nanoparticle (LNP) vaccine composition comprising: a. a mRNA nucleic acid with a N / P ratio of 4 to 7; b. a KC3-C15 (C8:1) ionizable cationic lipid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; c. cholesterol in a total amount of 25-40 mol% of the total lipid content of the LNP composition; d. a (L-Serine) PS lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition; e. DSPC phospholipid in a total amount of 5-25 mol% of the total lipid content of the LNP composition; and f. PEG-DMG in a total amount of 0-2.5 mol% of the total lipid content of the LNP composition. Embodiment 62: A nucleic acid lipid nanoparticle (LNP) vaccine composition comprising: a. a mRNA nucleic acid with a N / P ratio of 3 to 8; b. a KC3-C18 ionizable cationic lipid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; c. cholesterol in a total amount of 25-40 mol% of the total lipid content of the LNP composition; d. a (L-Serine) PS lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition; e. DSPC phospholipid in a total amount of 5-25 mol% of the total lipid content of the LNP composition; and 107 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 f. PEG-DMG in a total amount of 0-2.5 mol% of the total lipid content of the LNP composition. Embodiment 63: The composition of any one of embodiments 59-62, wherein the nucleic acid is a mRNA of SEQ ID NO: 2. Embodiment 64: Use of a (L-Serine) PS lipid in combination with an ionizable cationic lipid of any one of embodiments 1-9 in the LNP for targeting of the LNP to dendritic cells. Embodiment 65: The use of embodiment 64, wherein the LNP comprises mRNA. Embodiment 66: The use of any one of embodiments 64-65, wherein the LNP further comprises cholesterol. Embodiment 67: The use of embodiment 66, wherein the total amount of (L-Serine) PS lipid in the LNP is 2.5-10 mol% of the total lipid content of the LNP composition. Embodiment 68: The use of embodiment 67, wherein the LNP further comprises one or more additional phospholipids including DSPC. Embodiment 69: The use of embodiment 68, wherein the LNP further comprises a conjugated lipid. Embodiment 70: The use of embodiment 64, wherein the LNP comprises: a. a mRNA nucleic acid with a N / P ratio of 3 to 8; b. a KC3-PA or KC3-C17 (C8:1) ionizable cationic lipid (ICL), in a total amount of 40- 65 mol% of the total lipid content of the LNP composition; c. cholesterol in a total amount of 25-40 mol% of the total lipid content of the LNP composition; d. a (L-Serine) PS lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition; e. DSPC phospholipid in a total amount of 5-25 mol% of the total lipid content of the LNP composition; and f. a conjugated lipid in a total amount of 0-2.5 mol% of the total lipid content of the LNP composition. Embodiment 71: The use of embodiment 70, wherein the ICL is KC3-PA. Embodiment 72: The use of embodiment 70, wherein the ICL is KC3-C17 (C8:1). Embodiment 73: A lipid nanoparticle (LNP) composition comprising an ionizable lipid having the chemical structure: 108 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 , 1, 2, 3 or 4; R2and R3are each independently methyl; and n is an integer equal to 2 or 3. Embodiment 74: The composition of embodiment 73, wherein a is 0. Embodiment 75: The composition of embodiment 74, wherein b is 1. Embodiment 76: The composition of embodiment 74, wherein b is 3. Embodiment 77: The composition of embodiment 73, wherein a is 1. Embodiment 78: The composition of embodiment 77, wherein b is 1. Embodiment 79: The composition of embodiment 77, wherein b is 3. Embodiment 80: The composition of any one of embodiments 73-79, wherein n is 2. Embodiment 81: The composition of any one of embodiments 73-79, wherein n is 3. Embodiment 82: The composition of any one of embodiments 17-28, wherein the composition comprises an anionic lipid selected from the group consisting of: phosphatidylglycerol (PG), phosphatidic acid (PA), N-glutaryl-phosphatidylethanolamine (N-Glu-PE), N-succinyl- phosphatidylethanolamine (N-Suc-PE), and cardiolipin. Distearoylphosphatidylglycerol (DSPG), dipalmitoyphosphatidylglycerol (DPPG), N-succinyl-distearoylphosphatidylethanolamine (N- Suc-DSPE), N-glutaryl-distearoylphosphatidylethanolamine (N-glu-DSPE), distearoylphosphatidic acid (DSPA), and cardiolipin. Embodiment 83: The composition of any one of embodiments 17-28 or 82, wherein the composition comprises an anionic targeting phospholipid other than phosphatidyl-L-serine. Embodiment 84: The composition of any one of embodiments 17-28 or 82-83, wherein the composition comprises an anionic phospholipid selected from the group consisting of: DSPG and DPPG. 109 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 Embodiment 85: The composition of any one of embodiments 17-28 or 82-83, wherein the composition comprises an anionic phospholipid selected from the group consisting of: N-Glu- DSPE and N-Suc-DSPE. Embodiment 86: The composition of any one of embodiments 17-28 or 82-83, wherein the composition comprises a DSPA anionic phospholipid. Embodiment 87: The composition of any one of embodiments 17-28 or 82-83, wherein the composition comprises a Cardiolipin anionic phospholipid. Embodiment 88: The composition of any one of embodiments 1-63 or 73-87, wherein the ionizable lipid has the chemical structure: . one of embodiments 64-72, wherein the ionizable lipid has the chemical structure: . salt of a composition of an anionic phospholipid of Formula (V-A-1), having the chemical structure: wherein X+is an ammonium cation or a sodium (Na+) cation; and 110 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 a is 14, 15 or 16. Embodiment 91: The composition of embodiment 90, wherein a is 14 or 16. Embodiment 92: The composition of any one of embodiments 90 or 91, wherein X+is ammonium cation (NH4+). Embodiment 93: The composition of any one of embodiments 90 or 91, wherein X+is sodium cation (Na+) Embodiment 94: The composition of any one of embodiments 90-93, wherein the anionic phospholipid of Formula (V-A-1) is a sodium salt of distearoylphosphatidyl-L-serine (DSPS L- isomer). Embodiment 95: The composition of any one of embodiments 90-93, wherein the anionic phospholipid of Formula (V-A-1) is an ammonium salt of distearoylphosphatidyl-L-serine (DSPS L-isomer). Embodiment 96: The composition of any one of embodiments 90-93, wherein the anionic phospholipid of Formula (V-A-1) is a sodium salt of DPPS (L-isomer). Embodiment 97: The composition of any one of embodiments 90-93, wherein the anionic phospholipid of Formula (V-A-1) is a ammonium salt of DPPS (L-isomer). Embodiment 98: Use of the salt form composition of any one of embodiments 90-97 in the preparation of a liposomal nanoparticle (LNP) composition. Embodiment 99: The use of embodiment 98, in combination with one or more of the following LNP components during the preparation of the LNP composition: a mRNA nucleic acid; an ionizable cationic lipid (ICL); cholesterol; a (L-Serine) PS lipid; one or more phospholipids; and a conjugated lipid. Embodiment 100: The use of embodiment 98, comprising the step of combining the ammonium or salt form of a compound of Formula (V-A-1) with one or more of the following LNP components during the preparation of the LNP composition: a mRNA nucleic acid; an ionizable cationic lipid (ICL) of any one of embodiments 1-9 or 73-88; 111 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 cholesterol; a (L-Serine) PS lipid; one or more phospholipids; and a conjugated lipid. Embodiment 101: The use of any one of embodiments 98-100, wherein the LNP is a nucleic acid lipid nanoparticle vaccine composition comprising: a. a mRNA nucleic acid with a N / P ratio of 4 to 7; b. an ionizable cationic lipid of any one of embodiments 1-9 or 73-88 in a total amount of 40-65 mol% of the total lipid content of the LNP composition; c. cholesterol in a total amount of 25-40 mol% of the total lipid content of the LNP composition; d. a (L-Serine) PS lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition; e. DSPC phospholipid in a total amount of 5-25 mol% of the total lipid content of the LNP composition; and f. PEG-DMG in a total amount of 0-2.5 mol% of the total lipid content of the LNP composition. Embodiment 102: The use of embodiment 101, wherein the nucleic acid is mRNA of SEQ ID NO: 2. Embodiment 103: The composition of any one of embodiments 17-63, or 73-89, wherein LNP composition comprises the ionizable cationic lipid in a total amount of 46-65 mol% of the total lipid content of the LNP composition. Embodiment 104: The composition of any one of embodiments 17-63, 73-89, or 103, wherein the LNP composition comprises the PS in a total amount of about 5 mol% of the total lipid in the composition. Embodiment 105: The composition of any one of embodiments 17-63, 73-89 or 103-104, wherein the LNP composition comprises the conjugated lipid in a total amount of about 1.5 mol% of the total lipid content of the LNP composition. Embodiment 106: The composition of any one of embodiments 82-87, wherein the conjugated lipid is PEG-DMG; and the PS lipid is selected from the group consisting of: DSPS (L-isomer) and DPPS. 112 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 Embodiment 107: The composition of any one of embodiments 82-87, wherein the ionizable cationic lipid is one or more compounds selected from the group consisting of: KC3-OA, KC3- PA, KC3-C17 (C8:1), and KC3-C15 (C8:1). Embodiment 108: The composition of any one of embodiments 82-87, wherein the ionizable cationic lipid is KC3-PA. Embodiment 109: The composition of any one of embodiments 82-87, wherein the ionizable cationic lipid is KC3-OA. Embodiment 110: The composition of any one of embodiments 82-87, wherein the ionizable cationic lipid is KC3-C17 (C8:1). Embodiment 111: Use of a (L-Serine) PS lipid in combination with an ionizable cationic lipid of any one of embodiments 73-87 in the LNP for targeting of the LNP to dendritic cells. Embodiment 112: The use of embodiment 111, wherein the LNP comprises mRNA. Embodiment 113: The use of any one of embodiments 111-112, wherein the LNP further comprises cholesterol. Embodiment 114: The use of embodiment 113, wherein the total amount of (L-Serine) PS lipid in the LNP is 2.5-10 mol% of the total lipid content of the LNP composition. Embodiment 115: The use of embodiment 114, wherein the LNP further comprises one or more additional phospholipids including DSPC. Embodiment 116: The use of embodiment 115, wherein the LNP further comprises a conjugated lipid. Embodiment 117: The use of embodiment 111, wherein the LNP comprises: a. a mRNA nucleic acid with a N / P ratio of 3 to 8; b. a KC3-PA or KC3-C17 (C8:1) ionizable cationic lipid (ICL), in a total amount of 40- 65 mol% of the total lipid content of the LNP composition; c. cholesterol in a total amount of 25-40 mol% of the total lipid content of the LNP composition; d. a (L-Serine) PS lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition; e. DSPC phospholipid in a total amount of 5-25 mol% of the total lipid content of the LNP composition; and 113 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 f. a conjugated lipid in a total amount of 0-2.5 mol% of the total lipid content of the LNP composition. Embodiment 118: The use of embodiment 117, wherein the ICL is KC3-PA. Embodiment 119: The use of embodiment 117, wherein the ICL is KC3-C17 (C8:1). Embodiment 120: The composition of any one of embodiments 90-97, wherein X is an ammonium cation selected from the group consisting of: ammonium (NH4+), an alkylammonium, a dialkylammonium, and a trialkylammonium salt. Embodiment 121: The composition of embodiment 120, wherein X is X is an ammonium cation selected from the group consisting of: ammonium, dimethylamine, diethylamine, triethylamine, trimethylamine, 2-(dimethyamino)ethanol, diethanolamine, 2-(diethyamino)ethanol, ethanolamine, ethylenediamine, N-methyl-glucamine, imidazole, histidine, lysine, arginine, 4-(2- hydroxyethyl)-morpholine, piperazine, 1-(2-hydroxyethyl)-pyrrolidine, triethanolamine, and tromethamine (tris(hydroxymethyl)aminomethane). Embodiment 122: The composition of any one of embodiments 17-28 or 82-83, wherein the composition comprises an anionic phospholipid selected from the group consisting of: DSPG and DPPG, in a total amount of 2.5-7.5% of the total lipid content of the LNP composition. Embodiment 123: The composition of embodiment 122, wherein the composition comprises DSPG anionic phospholipid in a total amount of 2.5-7.5% of the total lipid content of the LNP composition. Embodiment 124: The composition of embodiment 122, wherein the composition comprises DPPG anionic phospholipid in a total amount of 2.5-7.5% of the total lipid content of the LNP composition. Embodiment 125: The use of embodiment 114, wherein the LNP further comprises one or more additional phospholipids including DSPC. Embodiment 126: A nucleic acid lipid nanoparticle (LNP) composition comprising: a. a nucleic acid; b. a KC3 ionizable cationic lipid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; c. cholesterol in a total amount of 23.5 - 43.5 mol% of the total lipid content of the LNP composition; 114 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 d. a (L-Serine) PS lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition; e. DSPC or HSPC phospholipid in a total amount of 5-25 mol% of the total lipid content of the LNP composition; and f. a PEG-containing conjugated lipid in a total amount of 0.5 mol% to 2.5 mol% of the total lipid content of the LNP composition. Embodiment 127: The composition of embodiment 126, wherein the nucleic acid is mRNA. Embodiment 128: The composition of any one of v 126-127, wherein the N / P ratio is 3 to 8. Embodiment 129: The composition of any one of embodiments 126-127, wherein the KC3 ionizable cationic lipid is selected from the group consisting of: KC3-OA, KC3-PA, KC3-C17 (8:1), and KC3-C15 (C8:1). Embodiment 130: The composition of embodiment 129, wherein the KC3 ionizable cationic lipid is KC3-OA. Embodiment 131: The composition of embodiment 129, wherein the KC3 ionizable cationic lipid is KC3-PA. Embodiment 132: The composition of embodiment 129, wherein the KC3 ionizable cationic lipid is KC3-C17(C8:1). Embodiment 133: The composition of embodiment 129, wherein the KC3 ionizable cationic lipid is KC3-C15(C8:1). Embodiment 134: The composition of any one of embodiments 126-133, wherein the conjugated lipid is PEG-DMG or PEG-DSG. Embodiment 135: The composition of embodiment 134, wherein the composition comprises the PEG-containing conjugated lipid in a total amount of 0.5 –2.0 mol% of the total lipid content of the LNP composition. Embodiment 136: The composition of any one of embodiments 126-135, wherein the composition comprises the KC3 ionizable cationic lipid in a total amount of 48 mol% of the total lipid content of the LNP composition. Embodiment 137: The composition of any one of embodiments 126-136, wherein the composition comprises DSPC and DSPS in a total amount of 10 mol% of the total lipid content of the LNP composition. 115 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 Embodiment 138: The composition of any one of embodiments 126-137, wherein the composition comprises 5 % DSPC or HSPC in a total amount of 5 mol% of the total lipid content of the LNP composition. Embodiment 139: The composition of any one of embodiments 126-137, wherein the composition comprises PEG-DMG in a total of 1.5 mol % of the total lipid content of the LNP composition. Embodiment 140: The composition of any one of embodiments 126-137, wherein the composition comprises cholesterol in a total amount of 40.5 mol % cholesterol of the total lipid content of the LNP composition. Embodiment 141: The composition of any one of embodiments 126-137, wherein the composition comprises the DSPC phospholipid in a total amount of 10 mol% of the total lipid content of the LNP composition. Embodiment 142: The composition of any one of embodiments 126-141 wherein the PEG- containing conjugated lipid is PEG2000-DMG. Embodiment 143: The composition of any one of embodiments 126-139, wherein the composition comprises the cholesterol in a total amount of 23.5 mol% of the total lipid content of the LNP composition. Embodiment 144: The composition of any one of embodiments 126-139, wherein the composition comprises the cholesterol in a total amount of 33.5 mol% of the total lipid content of the LNP composition. Embodiment 145: The composition of any one of embodiments 126-139, wherein the composition comprises the cholesterol in a total amount of 38.5 mol% of the total lipid content of the LNP composition. Embodiment 146: The composition of any one of embodiments 126-139, wherein the composition comprises the cholesterol in a total amount of 40.5 mol% of the total lipid content of the LNP composition. Embodiment 147: The composition of any one of embodiments 126-139, wherein the composition comprises the cholesterol in a total amount of 42.7 mol% of the total lipid content of the LNP composition. Embodiment 148: The composition of any one of embodiments 126-139, wherein the composition comprises the cholesterol in a total amount of 43.5 mol% of the total lipid content of the LNP composition. 116 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 Embodiment 149: The composition of any one of embodiments 126-139, wherein the composition comprises the cholesterol in a total amount of 33.5-43.5 mol% of the total lipid content of the LNP composition. Embodiment 150: The composition of any one of embodiments 126-149, wherein the composition comprises the KC3 ionizable cationic lipid in a total amount of 45-55 mol% of the total lipid content of the LNP composition. Embodiment 151: A nucleic acid lipid nanoparticle (LNP) composition comprising: a. a mRNA nucleic acid; b. a KC3 ionizable cationic lipid selected from the group consisting of: KC3-OA, KC3- PA, KC3-C17 (8:1), and KC3-C15 (C8:1), in a total amount of 45-55 mol% of the total lipid content of the LNP composition; c. cholesterol in a total amount of 33.5-43.5 mol% of the total lipid content of the LNP composition; d. a (L-Serine) DPPS lipid in a total amount of 5 mol% of the total lipid content of the LNP composition; e. DSPC or HSPC phospholipid in a total amount of 5 mol% of the total lipid content of the LNP composition; and f. a PEG-DMG conjugated lipid in a total amount of 1.5 mol% of the total lipid content of the LNP composition. Embodiment 152: A lipid nanoparticle (LNP) composition comprising a KC3 ionizable cationic lipid, a (L-Serine) PS lipid, cholesterol, one or more phospholipids comprising at least one anionic phospholipid, and a conjugated lipid, wherein the LNP is obtained by a process comprising the step of dissolving a sodium or ammonium salt of the anionic phospholipid. Embodiment 153: The composition of embodiment 152, wherein the anionic phospholipid is the salt of any one of embodiments 90-97. Embodiment 154: The composition of any one of embodiments 152-153, wherein the composition comprises a nucleic acid. Embodiment 155: The composition of embodiment 154, wherein the nucleic acid is mRNA. Embodiment 156: The composition of embodiment 155, wherein the composition is a vaccine. Embodiment 157: The composition of any one of embodiments 152-156, wherein the total amount of phospholipids in the composition is 5-25 mol% of the total lipid content of the LNP 117 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 composition, and the total amount of the phosphatidylserine (PS) is 2.5-10 mol% of the total lipid content of the LNP composition; and the total amount of the conjugated lipid in the composition is a total amount of 0.5 – 2.5 mol% of the total lipid content of the LNP composition. Embodiment 158: The composition of any one of embodiments 152-156, wherein the composition comprises 48 mol% of the KC3 ionizable cationic lipid, 40.5 mol% cholesterol, and 5 mol% (L-Serine) DPPS lipid, wherein each mol% refers to the mol% of the total lipid content of the LNP composition. Embodiment 159: The composition of any one of embodiments 152-156, wherein the composition comprises 48 mol% of the KC3 ionizable cationic lipid, 38.5 mol% cholesterol, and 5 mol% (L-Serine) DPPS lipid, wherein each mol% refers to the mol% of the total lipid content of the LNP composition. Embodiment 160: The composition of any one of embodiments 152-156, wherein the composition comprises 46-54 mol% of the KC3 ionizable cationic lipid, and 5 mol% (L-Serine) DPPS lipid, wherein each mol% refers to the mol% of the total lipid content of the LNP composition. Embodiment 161: The composition of any one of embodiments 152-156, wherein the composition comprises 45 mol% of the KC3 ionizable cationic lipid, 42.7 mol% cholesterol, and 5 mol% (L-Serine) DPPS lipid, wherein each mol% refers to the mol% of the total lipid content of the LNP composition. Embodiment 162: The composition of any one of embodiments 152-156, wherein the composition comprises 50 mol% of the KC3 ionizable cationic lipid, 38.5 mol% cholesterol, 5 mol% (L-Serine) DPPS lipid, and 118 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 a total of 10 mol% phospholipid concentration; wherein each mol% refers to the mol% of the total lipid content of the LNP composition. Embodiment 163: The composition of any one of embodiments 152-156, wherein the composition comprises 48 mol% of the KC3 ionizable cationic lipid, 40.5 mol% cholesterol, 5 mol% (L-Serine) DPPS lipid, and a total of 10 mol% phospholipid concentration; wherein each mol% refers to the mol% of the total lipid content of the LNP composition. Embodiment 164: The composition of any one of embodiments 152-156, wherein the composition comprises 48 mol% of the KC3 ionizable cationic lipid, 40.5 mol% cholesterol, 5 mol% (L-Serine) DPPS lipid, 5 mol% DSPC or DPPC; and a total of 10 mol% phospholipid concentration; wherein each mol% refers to the mol% of the total lipid content of the LNP composition. Embodiment 165: The composition of any one of embodiments 152-156, wherein the composition comprises 46.5 mol% of the KC3 ionizable cationic lipid, 42 mol% cholesterol, 5 mol% (L-Serine) DPPS lipid, wherein each mol% refers to the mol% of the total lipid content of the LNP composition. Embodiment 166: The composition of any one of embodiments 158-165, wherein the composition further comprises a total of 5 mol% DSPC or HSPC of the total lipid content of the LNP composition. Embodiment 167: The composition of any one of embodiments 158-166, wherein the composition further comprises a total of 1.5 mol% PEG-DMG of the total lipid content of the LNP composition. Embodiment 168: The composition of any one of embodiments 158-163, wherein the composition comprises a total of 10 mol% of DSPC / DPPC phospholipid of the total lipid content of the LNP composition. 119 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 Embodiment 169: A phosphatidylserine salt selected from the group consisting of DSPS sodium, DPPS sodium, DSPS ammonium and DPPS ammonium. Embodiment 170: Use of a DSPS-Na salt or a DPPS-NH4+salt in the preparation of a LNP comprising a (L-Serine) PS lipid, a sterol, a conjugated lipid, a phospholipid for targeting the LNP to dendritic cells. Embodiment 171: A solution comprising ethanol and DSPS or DPPS, the solution obtained by a process comprising the step of dissolving a phosphatidylserine salt in ethanol, wherein the phosphatidylserine salt is selected from the group consisting of DSPS sodium, DPPS sodium, DSPS ammonium and DPPS ammonium. Aspects of the disclosure relate to a lipid nanoparticle (LNP) vaccine composition comprising: (a) a nucleic acid; (b) an ionizable cationic lipid at a N / P ratio of 3 to 8 relative to the nucleic acid wherein the ionizable lipid has the chemical structure: NOR1, of C15-C19containing one or two olefins, andwherein the ionizable cationic lipid is present in the LNP vaccine composition in a total amount of 40-65 mol% of the total lipid content of the LNP composition, and the ionizable cationic lipid is optionally selected from the group DLin-KC3-DMA, KC3-01, KC3-OA, KC3-PA, KC3-C17 (8:1), KC3- C15 (C8:1) Compound 3 (Table 1A), Compound 8 (Table 1A); (c) a sterol in a total amount of 25- 45 mol% of the total lipid content of the LNP composition; (d) one or more phospholipids in a total amount of phospholipids of 5-25 mol% of the total lipid content of the LNP composition; and (e) a conjugated lipid in a total amount of 0.5 – 2.5 mol% of the total lipid content of the LNP composition. In some embodiments, the one or more phospholipids comprises a phosphatidylserine (PS) lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition. In some embodiments, the PS-lipid is DSPS or DPPS. In some embodiments, the second phospholipid is selected from the group consisting of: DSPC, HSPC, DPPC, and sphingomyelin. 120 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 In some embodiments, the ionizable cationic lipid comprises either monounsaturated alkyl chains or di-unsaturated alkyl chains, wherein the olefins are separated by at least two methylene groups. In some embodiments, the ionizable cationic lipid is selected from the group consisting of: KC3-01, KC3-OA, KC3-PA, KC3-C17 (8:1), KC3-C15 (C8:1), and Compound 8 (Table 1A). In some embodiments, the composition has an N / P ratio of 5-6 relative to the nucleic acid. In some embodiments, the nucleic acid is an RNA. In some embodiments, the nucleic acid is mRNA. In some embodiments, the nucleic acid is a chemically modified RNA. In some embodiments, the nucleic acid is modified with N-methylpseudouridine. Other aspects of the disclosure relate to a lipid nanoparticle (LNP) composition comprising: (a) a nucleic acid; (b) an ionizable cationic lipid at a N / P ratio of 3 to 8 relative to the nucleic acid wherein the ionizable lipid has the chemical structure: NOR1, wherein containing one or two olefins, and wherein the ionizable cationic lipid is present in the LNP composition in a total amount of 40-65 mol% of the total lipid content of the LNP composition, and the ionizable cationic lipid is optionally selected from the group Compound 3 (Table 1A), Compound 8 (Table 1A), DLin-KC3-DMA, KC3-01, KC3-OA, KC3-PA, KC3-C17 (8:1), and KC3-C15 (C8:1); (c) a sterol in a total amount of 25-45 mol% of the total lipid content of the LNP composition; (d) one or more phospholipids in a total amount of phospholipids of 5-25 mol% of the total lipid content of the LNP composition, and comprising a dipalmitoylphosphatidylserine (DPPS) lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition; and (e) a conjugated lipid in a total amount of 0.5 – 2.5 mol% of the total lipid content of the LNP composition. In some embodiments, a second phospholipid is selected from the group consisting of: DSPC, HSPC, and sphingomyelin. In some embodiments, the ionizable cationic lipid is at an N / P ratio of 5-6 relative to the nucleic acid. 121 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 In some embodiments, the ionizable cationic lipid contains either two monounsaturated alkyl chains or two di-unsaturated alkyl chains, and wherein the olefins are separated by at least two methylene groups. In some embodiments, the ionizable cationic lipid is selected from the group KC3-01, KC3- OA, KC3-PA, KC3-C17 (8:1), KC3-C15 (C8:1), and Compound 8 (Table 1A). In some embodiments, the ionizable cationic lipid is 3-((S)-2,2-di((Z)-octadec-9-en-1-yl)- 1,3-dioxolan-4-yl)-N,N-dimethylpropan-1-amine (KC3-OA(S)). In some embodiments, the nucleic acid is an mRNA. In some embodiments, the mRNA is chemically modified with N-methylpseudouridine. Aspects of the disclosure relate to a nucleic acid lipid nanoparticle (LNP) composition comprising: (a) a nucleic acid; (b) an ionizable cationic lipid at a N / P ratio of 3 to 8 relative to the nucleic acid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; (c) a sterol in a total amount of 25-45 mol% of the total lipid content of the LNP composition; (d) one or more phospholipids in a total amount of phospholipids of 5-25 mol% of the total lipid content of the LNP composition; and (e) a conjugated lipid in a total amount of 0.5 – 2.5 mol% of the total lipid content of the LNP composition comprising a PEG-lipid having a poly(ethylene glycol) chain terminally attached to a linking moiety and two hydrocarbon chains terminally attached to the same linking moiety, wherein the hydrocarbon chains are saturated C12-chains independently selected from n-dodecyl (lauryl) group and n-dodecanoyl (lauroyl) group. In some embodiments, the linking moiety is glyceryl group, N-oxycarbonyl glycerophoshoryl ethanolaminocarbonyl group, oxycarbonylamide group, or oxyacetamide group. In some embodiments, the poly(ethyene glycol) chain is methoxy-poly(ethyene glycol) with an average molecular weight of 2000. In some embodiments, the PEG-lipid is mPEG-1,2-dilauroylglycerol (PEG-DLG), mPEG-1,2-dilaurylglycerol (PEG-DLG), PEG-1,2-dilaurylglycerol, PEG-DLPE, PEG-oxycarbonyl-N,N-didodecylamide, or mPEG-N,N-didodecylacetamide. In some embodiments, the LNP has the z-average particle size of 60-150 nm and is freeze- thaw stable. In some embodiments, the one or more phospholipids comprise a phosphatidylserine (PS) lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition, and wherein the LNP composition comprises a phosphatidylserine (PS) lipid in a total amount of 2.5- 10 mol% of the total lipid content of the LNP composition. 122 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 Aspects of the disclosure relate to a nucleic acid lipid nanoparticle (LNP) human vaccine composition comprising: (a) a nucleic acid; (b) an ionizable cationic lipid at a N / P ratio of 3 to 8 relative to the nucleic acid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; (c) a sterol in a total amount of 25-45 mol% of the total lipid content of the LNP composition; (d) one or more phospholipids in a total amount of phospholipids of 5-25 mol% of the total lipid content of the LNP composition, and comprising a phosphatidylglycerol (PG) in a total amount of 1.0 – 10 mol% of the total lipid content of the LNP composition; and (e) a conjugated lipid in a total amount of 0.5 – 2.5 mol% of the total lipid content of the LNP composition. In some embodiments, the ionizable cationic lipid is selected from the group consisting of: KC3-01, KC3-OA, KC3-PA, KC3-C17 (8:1), KC3-C15 (C8:1), and Compound 8 (Table 1A). In some embodiments, the sterol is cholesterol; and the phosphatidylglycerol (PG) is an anionic phospholipid selected from the group consisting of: distearoylphosphatidylglycerol (DSPG) and dipalmitoyphosphatidylglycerol (DPPG). In some embodiments, the nucleic acid is mRNA and the ionizable cationic lipid is present at a N / P ratio of 4 to 7 relative to the nucleic acid. In some embodiments, the conjugated lipid is selected from PEG-DMG, PEG-DLG, and PEG-DLPE. In some embodiments, the one or more phospholipids comprise a phospholipid selected from the group consisting of: distearoylphosphatidylcholine (DSPC) and hydrogenated soy phosphatidylcholine (HSPC). Aspects of the disclosure relate to a method of making a nucleic acid delivery composition comprising lipids wherein the lipids comprise phosphatidylserine, the method comprising a step of dissolving the phosphatidylserine in ethanol wherein the phosphatidylserine is in the form of an ammonium salt of the phosphatidylserine. In some embodiments, the phosphatidylserine is DPPS. In some embodiments, the phosphatidylserine is dissolved in ethanol to the concentration of more than 0.2 mM. In some embodiments, the ammonium salt is a salt comprising an ammonium selected from the group consisting of: ammonium, alkyammonium, dialkylammonium, trialkylammonium, and tetraalkylammonium. In some embodiments, the ammonium form selected from the group consisting of: ammonia, dimethylamine, diethylamine, triethylamine, trimethylamine, 2- 123 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 (dimethyamino)ethanol, diethanolamine, 2-(diethyamino)ethanol, ethanolamine, ethylenediamine, N-methyl-glucamine, imidazole, histidine, lysine, arginine, 4-(2-hydroxyethyl)-morpholine, piperazine, 1-(2-hydroxyethyl)-pyrrolidine, triethanolamine, or tromethamine (tris(hydroxymethyl)aminomethane). Aspects of the disclosure relate to a method for delivery of a nucleic acid into cells comprising the step of: contacting the lipid nanoparticle (LNP) with the cells, wherein the cells are human dendritic cells, wherein the LNP composition was obtained by a process of aspects of the disclosure. In some embodiments, the LNP comprises an ionizable cationic lipid of the disclosure. In some embodiments, the cells are in a human or animal subject. In some embodiments, the LNP is administered to the subject intramuscularly, subcutaneously, intradermally, or topically. Aspects of the disclosure relate to an LNP composition comprising: (a) a nucleic acid, wherein the nucleic acid is an mRNA; (b) a cholesterol sterol in a total amount of 25-40 mol% of the total lipid content of the LNP composition; (c) an ionizable cationic lipid having a N / P ratio of 3 to 8 relative to the nucleic acid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; (d) one or more phospholipids in a total amount of phospholipids of 5-25 mol% of the total lipid content of the LNP composition, the one or more phospholipids are selected from the group consisting of (i) an ammonium salt of dipalmitoylphosphatidyl-L-serine ((L- serine)DPPS) lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition; and (ii) a distearoylphosphatidylcholine (DSPC) phospholipid in a total amount of 5-25 mol% of the total lipid content of the LNP composition; and (e) a PEG-containing conjugated lipid in a total amount of 0.5 – 2.5 mol% of the total lipid content of the LNP composition. Aspects of the disclosure relate to a method for administering a nucleic acid to a subject in need thereof comprising administering a nucleic acid lipid nanoparticle (LNP) composition to a human subject, the nucleic acid LNP composition comprising: (a) a nucleic acid; (b) an ionizable cationic lipid at a N / P ratio of 3 to 8 relative to the nucleic acid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; (c) a sterol in a total amount of 25-45 mol% of the total lipid content of the LNP composition; (d) one or more phospholipids in a total amount of phospholipids of 5-25 mol% of the total lipid content of the LNP composition, and comprising a phosphatidylglycerol (PG) in a total amount of 1.0 – 10 mol% of the total lipid content of the LNP 124 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 composition; and (e) a conjugated lipid in a total amount of 0.5 – 2.5 mol% of the total lipid content of the LNP composition. Aspects of the disclosure relate to a nucleic acid lipid nanoparticle (LNP) composition comprising: (a) a nucleic acid; (b) an ionizable cationic lipid at a N / P ratio of 3 to 8 relative to the nucleic acid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; (c) a sterol in a total amount of 25-45 mol% of the total lipid content of the LNP composition; (d)one or more phospholipids in a total amount of phospholipids of 5-25 mol% of the total lipid content of the LNP composition, and comprising a phosphatidylserine (PS) lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition; and (e) a conjugated lipid in a total amount of 0.5 – 2.5 mol% of the total lipid content of the LNP composition. In some embodiments, the nucleic acid is mRNA; the ionizable cationic lipid is present in the LNP composition at a N / P ratio of 4 to 7 relative to the nucleic acid; the sterol is cholesterol; and the conjugated lipid is a PEG-containing conjugated lipid. In some embodiments, the one or more phospholipids comprise at least two phospholipids having mismatched acyl chain lengths. In some embodiments, the phosphatidylserine (PS) lipid is dipalmitoylphosphatidyl-L- serine ((L-serine)DPPS). In some embodiments, the one or more phospholipids comprise a phospholipid selected from the group consisting of: distearoylphosphatidylcholine (DSPC) and hydrogenated soy phosphatidylcholine (HSPC). In some embodiments, the one or more phospholipids consist of distearoylphosphatidylcholine (DSPC) and dipalmitoylphosphatidyl-L-serine ((L-serine)DPPS). In some embodiments, the PEG-containing conjugated lipid is PEG(2000)-dimyristoylglycerol (PEG-DMG). Aspects of the disclosure relate to an LNP composition wherein the ionizable lipid has the chemical structure: Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 , 1, 2, 3 or 4; are with hydroxyl; and n is an integer equal to 2, 3 or 4. In some embodiments, R2and R3are each methyl; and n is 3 or 4. In some embodiments, the ionizable cationic lipid is one or more compounds selected from the group consisting of: KC3-OA, KC3-PA, KC3-C17 (8:1), and KC3-C15 (C8:1). In some embodiments, the ionizable cationic lipid is KC3-PA. In some embodiments, the ionizable cationic lipid is KC3-OA. In some embodiments, the ionizable cationic lipid is KC3-C17 (C8:1). Aspects of the disclosure relate to an ionizable cationic lipid selected from the group consisting of KC3-PA, KC3-C17 (8:1), and KC3-C15 (C8:1). Aspects of the disclosure relate to a nucleic acid lipid nanoparticle (LNP) composition comprising: (a) a nucleic acid; (b) an ionizable cationic lipid at a N / P ratio of 3 to 8 relative to the nucleic acid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; (c) a sterol in a total amount of 25-45 mol% of the total lipid content of the LNP composition; (d) one or more phospholipids in a total amount of phospholipids of 5-25 mol% of the total lipid content of the LNP composition, and comprising a phosphatidylserine (PS) lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition; and (d) a conjugated lipid in a total amount of 0.5 – 2.5 mol% of the total lipid content of the LNP composition. In some embodiments, the sterol is cholesterol. In other embodiments, the sterol is beta- sitosterol. In some embodiments, the one or more phospholipids comprise phospholipids having mismatched acyl chain lengths. In some embodiments, the PS lipid is dipalmitoylphosphatidyl-L- serine ((L-serine)DPPS). In some embodiments, the one or more phospholipids comprise a phospholipid selected from the group consisting of: distearoylphosphatidylcholine (DSPC) and hydrogenated soy phosphatidylcholine (HSPC). In some embodiments, the one or more phospholipids comprise distearoylphosphatidylcholine (DSPC) and the phosphatidylserine (PS). In some embodiments, the ionizable cationic lipid is 3-((S)-2,2-di((Z)-octadec-9-en-1-yl)-1,3- dioxolan-4-yl)-N,N-dimethylpropan-1-amine (KC3-OA). In some embodiments, the ionizable cationic lipid is 3-((S)-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N-dimethylpropan-1- amine (KC3-OA). In some embodiments, the one or more phospholipids consist of 126 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 distearoylphosphatidylcholine (DSPC) and dipalmitoylphosphatidyl-L-serine ((L-serine)DPPS), and the phosphatidylserine (PS) is (L-Serine)DPPS. In some embodiments, the conjugated lipid is a PEG-containing conjugated lipid, and wherein the PEG-containing conjugated lipid is selected from the group consisting of: PEG(2000)- dimyristoylglycerol (PEG-DMG), 1,2-dilauroyl-sn-glycero-3-phosphoethanolamine-N- [methoxy(polyethylene glycol)-2000] (PEG-DLPE), and PEG(2000)-dilauroylglycerol (PEG- DLG). In some embodiments, the nucleic acid is mRNA; the one or more phospholipids comprise phosphatidylserine (PS) and one or more phospholipids selected from the group consisting of: distearoylphosphatidylcholine (DSPC), hydrogenated soy phosphatidylcholine (HSPC), and dipalmitoylphosphatidyl-L-serine ((L-serine)DPPS); and the conjugated lipid comprises a polyethylene glycol (PEG). In some embodiments, the conjugated lipid is PEG(2000)- dimyristoylglycerol (PEG-DMG). In some embodiments, the ionizable cationic lipid is 3-((S)-2,2- di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N-dimethylpropan-1-amine (KC3-OA). In some embodiments, the one or more phospholipids consist of distearoylphosphatidylcholine (DSPC) and dipalmitoylphosphatidyl-L-serine ((L-serine)DPPS). In some embodiments, the dipalmitoylphosphatidyl-L-serine ((L-serine)DPPS) is an ammonium salt of (L-Serine)DPPS. In some embodiments, (a) the nucleic acid is mRNA; (b) the sterol is a cholesterol sterol in a total amount of 25-45 mol% of the total lipid content of the LNP composition; (c) the ionizable cationic lipid is at a N / P ratio of 3 to 8 relative to the nucleic acid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; (d) the one or more phospholipids is in a total amount of phospholipids of 5-25 mol% of the total lipid content of the LNP composition, the one or more phospholipids consisting of: (i)dipalmitoylphosphatidyl-L-serine ((L-serine)DPPS) lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition; and (ii) a distearoylphosphatidylcholine (DSPC) phospholipid in a total amount of 5-25 mol% of the total lipid content of the LNP composition; (e) the conjugated lipid is a PEG-containing conjugated lipid in a total amount of 0.5 – 2.5 mol% of the total lipid content of the LNP composition. In some embodiments, the PEG-containing conjugated lipid is selected from the group consisting of: PEG(2000)-dimyristoylglycerol (PEG-DMG), 1,2-dilauroyl-sn-glycero-3- 127 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (PEG-DLPE), and PEG(2000)- dilauroylglycerol (PEG-DLG). Aspects of the disclosure relate to a nucleic acid lipid nanoparticle (LNP) human vaccine composition comprising: (a) a nucleic acid; (b) an ionizable cationic lipid at a N / P ratio of 3 to 8 relative to the nucleic acid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; (c) a sterol in a total amount of 25-45 mol% of the total lipid content of the LNP composition; (d) one or more phospholipids in a total amount of phospholipids of 5-25 mol% of the total lipid content of the LNP composition, and comprising a phosphatidylglycerol (PG) in a total amount of 1.0 – 10 mol% of the total lipid content of the LNP composition; and (e) a conjugated lipid in a total amount of 0.5 – 2.5 mol% of the total lipid content of the LNP composition. In some embodiments, the sterol is cholesterol; and the phosphatidylglycerol (PG) is an anionic phospholipid selected from the group consisting of: distearoylphosphatidylglycerol (DSPG) and dipalmitoyphosphatidylglycerol (DPPG). In some embodiments, the nucleic acid is mRNA and the ionizable cationic lipid is present at a N / P ratio of 4 to 7 relative to the nucleic acid. Aspects of the disclosure relate to a nucleic acid lipid nanoparticle (LNP) composition comprising an ionizable cationic lipid in a total amount of 40-65 mol% of the total lipid content of the LNP composition, wherein the ionizable cationic lipid is selected from the group consisting of: KC3-PA, KC3-C17 (8:1), and KC3-C15 (C8:1), KC3-OA, and KC3-01 : ; 128 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 ; ; composition comprising: (a) a nucleic acid; (b) an ionizable cationic lipid at a N / P ratio of 3 to 8 relative to the nucleic acid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; (c) a sterol in a total amount of 25-45 mol% of the total lipid content of the LNP composition; (d) one or more phospholipids in a total amount of phospholipids of 5-25 mol% of the total lipid content of the LNP composition; and (d) a conjugated lipid in a total amount of 0.5 – 2.5 mol% of the total lipid content of the LNP composition, wherein the conjugated lipid which is a PEG-lipid having a poly(ethylene glycol) chain terminally attached to a linking moiety and two hydrocarbon chains terminally attached to the same linking moiety, wherein the two hydrocarbon chains are saturated C12-chains independently selected from n-dodecyl (lauryl) group and n-dodecanoyl (lauroyl) group. 129 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 In some embodiments, the linking moiety is a glyceryl group, N-oxycarbonyl glycerophoshoryl ethanolaminocarbonyl group, oxycarbonylamide group, or oxyacetamide group. In some embodiments, the poly(ethyene glycol) chain is methoxy-poly(ethyene glycol) with an average molecular weight of 2000. In some embodiments, the PEG-lipid is mPEG-1,2- dilauroylglycerol (PEG-DLG), mPEG-1,2-dilaurylglycerol (PEG-DLG), PEG-1,2- dilaurylglycerol, PEG-DLPE, PEG-oxycarbonyl-N,N-didodecylamide, or mPEG-N,N- didodecylacetamide. Additional Embodiments Non-limiting embodiments are described below each of which is considered to be within the present disclosure. 1. A lipid nanoparticle (LNP) vaccine composition comprising: a. a nucleic acid; b. an ionizable cationic lipid at a N / P ratio of 3 to 8 relative to the nucleic acid wherein the NOionizable lipid has the chemical , wherein R1 isalkyl group of C15-C19containing one or wherein the ionizable cationic lipid is present in the LNP vaccine composition in a total amount of 40-65 mol% of the total lipid content of the LNP composition, and the ionizable cationic lipid is optionally selected from the group DLin-KC3-DMA, KC3-01, KC3-OA, KC3-PA, KC3-C17 (8:1), KC3-C15 (C8:1) Compound 3 (Table 1A), Compound 8 (Table 1A); c. a sterol in a total amount of 25-45 mol% of the total lipid content of the LNP composition; d. one or more phospholipids in a total amount of phospholipids of 5-25 mol% of the total lipid content of the LNP composition; and e. a conjugated lipid in a total amount of 0.5 – 2.5 mol% of the total lipid content of the LNP composition. 2. The composition of embodiment 1, wherein the one or more phospholipids comprises a phosphatidylserine (PS) lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition. 130 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 3. The composition of embodiment 2, wherein the PS-lipid is DSPS or DPPS. 4. The composition of embodiment 1, wherein a second phospholipid is selected from the group consisting of: DSPC, HSPC, DPPC, and sphingomyelin. 5. The LNP vaccine composition of embodiment 1, wherein the ionizable cationic lipid comprises either monounsaturated alkyl chains or di-unsaturated alkyl chains, wherein the olefins are separated by at least two methylene groups. 6. The composition of embodiment 1, wherein the ionizable cationic lipid is selected from the group consisting of: KC3-01, KC3-OA, KC3-PA, KC3-C17 (8:1), KC3-C15 (C8:1), and Compound 8 (Table 1A). 7. The composition of any one of embodiments 1-6, having an N / P ratio of 5-6 relative to the nucleic acid. 8. The composition of embodiment 7, wherein the nucleic acid is an RNA. 9. The composition of embodiment 8, wherein the nucleic acid is mRNA, optionally chemically modified RNA, and further optionally modified with N-methylpseudouridine. 10. A lipid nanoparticle (LNP) composition comprising: a. a nucleic acid; b. an ionizable cationic lipid at a N / P ratio of 3 to 8 relative to the nucleic acid wherein the ionizable lipid has the chemical structure: NO, wherein R1 is alkyl group of C15-C19 containing one or Wherein the ionizable cationic lipid is present in the LNP composition in a total amount of 40-65 mol% of the total lipid content of the LNP composition, and the ionizable cationic lipid is optionally selected from the group Compound 3 (Table 1A), Compound 8 (Table 1A), DLin-KC3-DMA, KC3-01, KC3-OA, KC3-PA, KC3-C17 (8:1), and KC3-C15 (C8:1); c. a sterol in a total amount of 25-45 mol% of the total lipid content of the LNP composition; 131 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 d. one or more phospholipids in a total amount of phospholipids of 5-25 mol% of the total lipid content of the LNP composition, and comprising a dipalmitoylphosphatidylserine (DPPS) lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition; and e. a conjugated lipid in a total amount of 0.5 – 2.5 mol% of the total lipid content of the LNP composition. 11. The composition of embodiment 10, wherein a second phospholipid is selected from the group consisting of: DSPC, HSPC, and sphingomyelin. 12. The composition of any one of embodiments 10-11, wherein the ionizable cationic lipid is at an N / P ratio of 5-6 relative to the nucleic acid. 13. The LNP composition of embodiment 12, wherein the ionizable cationic lipid contains either two monounsaturated alkyl chains or two di-unsaturated alkyl chains, and wherein the olefins are separated by at least two methylene groups. 14. The LNP composition of embodiment 13, wherein the ionizable cationic lipid is selected from the group KC3-01, KC3-OA, KC3-PA, KC3-C17 (8:1), KC3-C15 (C8:1), and Compound 8 (Table 1A). 15. The composition of embodiment 14, wherein the ionizable cationic lipid is 3-((S)-2,2-di((Z)- octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N-dimethylpropan-1-amine (KC3-OA). 16. The composition of any one of embodiments 10-15, wherein the nucleic acid is an mRNA. 17. The composition of embodiment 16, wherein the mRNA is chemically modified with N- methylpseudouridine. 18. A nucleic acid lipid nanoparticle (LNP) composition comprising: a. a nucleic acid; b. an ionizable cationic lipid at a N / P ratio of 3 to 8 relative to the nucleic acid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; c. a sterol in a total amount of 25-45 mol% of the total lipid content of the LNP composition; d. one or more phospholipids in a total amount of phospholipids of 5-25 mol% of the total lipid content of the LNP composition; and e. a conjugated lipid in a total amount of 0.5 – 2.5 mol% of the total lipid content of the LNP composition comprising a PEG-lipid having a poly(ethylene glycol) chain terminally attached to a linking moiety and two hydrocarbon chains terminally attached 132 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 to the same linking moiety, wherein the hydrocarbon chains are saturated C12-chains independently selected from n-dodecyl (lauryl) group and n-dodecanoyl (lauroyl) group. 19. The composition of embodiment 18, wherein the linking moiety is glyceryl group, N- oxycarbonyl glycerophoshoryl ethanolaminocarbonyl group, oxycarbonylamide group, or oxyacetamide group. 20. The composition of embodiment 19, wherein the poly(ethyene glycol) chain is methoxy- poly(ethyene glycol) with an average molecular weight of 2000. 21. The composition of embodiment 20, wherein the PEG-lipid is mPEG-1,2-dilauroylglycerol (PEG-DLG), mPEG-1,2-dilaurylglycerol (PEG-DLG), PEG-1,2-dilaurylglycerol, PEG- DLPE, PEG-oxycarbonyl-N,N-didodecylamide, or mPEG-N,N-didodecylacetamide. 22. The composition of any one of embodiments 18-21, wherein the LNP has the z-average particle size of 60-150 nm and is freeze-thaw stable. 23. The composition of any one of embodiments 18-22, wherein one or more phospholipids comprise a phosphatidylserine (PS) lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition, and wherein the LNP composition comprises a phosphatidylserine (PS) lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition. 24. A nucleic acid lipid nanoparticle (LNP) human vaccine composition comprising: a. a nucleic acid; b. an ionizable cationic lipid at a N / P ratio of 3 to 8 relative to the nucleic acid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; c. a sterol in a total amount of 25-45 mol% of the total lipid content of the LNP composition; d. one or more phospholipids in a total amount of phospholipids of 5-25 mol% of the total lipid content of the LNP composition, and comprising a phosphatidylglycerol (PG) in a total amount of 1.0 – 10 mol% of the total lipid content of the LNP composition; and e. a conjugated lipid in a total amount of 0.5 – 2.5 mol% of the total lipid content of the LNP composition. 25. The composition of embodiment 24, wherein the ionizable cationic lipid is selected from the group consisting of: KC3-01, KC3-OA, KC3-PA, KC3-C17 (8:1), KC3-C15 (C8:1), and Compound 8 (Table 1A). 26. The composition of embodiment 24, wherein 133 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 a. the sterol is cholesterol; and b. the phosphatidylglycerol (PG) is an anionic phospholipid selected from the group consisting of: distearoylphosphatidylglycerol (DSPG) and dipalmitoyphosphatidylglycerol (DPPG). 27. The composition of embodiment 24, wherein the nucleic acid is mRNA and the ionizable cationic lipid is present at a N / P ratio of 4 to 7 relative to the nucleic acid. 28. The composition of embodiment 24, wherein the conjugated lipid is selected from PEG-DMG, PEG-DLG, and PEG-DLPE. 29. The composition of embodiment 26, wherein the one or more phospholipids comprise a phospholipid selected from the group consisting of: distearoylphosphatidylcholine (DSPC) and hydrogenated soy phosphatidylcholine (HSPC). 30. A method of making a nucleic acid delivery composition comprising lipids wherein the lipids comprise phosphatidylserine, the method comprising a step of dissolving the phosphatidylserine in ethanol wherein the phosphatidylserine is in the form of an ammonium salt of the phosphatidylserine. 31. The method of embodiment 30, wherein the phosphatidylserine is DPPS. 32. The method of any one of embodiments 30-31, wherein the phosphatidylserine is dissolved in ethanol to the concentration of more than 0.2 mM. 33. The method of any one of embodiments 30-32, wherein the ammonium salt is a salt comprising an ammonium selected from the group consisting of: ammonium, alkyammonium, dialkylammonium, trialkylammonium, and tetraalkylammonium. 34. The method of any one of embodiments 30-32, wherein the ammonium is selected from the ammonium form selected from the group consisting of: ammonia, dimethylamine, diethylamine, triethylamine, trimethylamine, 2-(dimethyamino)ethanol, diethanolamine, 2- (diethyamino)ethanol, ethanolamine, ethylenediamine, N-methyl-glucamine, imidazole, histidine, lysine, arginine, 4-(2-hydroxyethyl)-morpholine, piperazine, 1-(2-hydroxyethyl)- pyrrolidine, triethanolamine, or tromethamine (tris(hydroxymethyl)aminomethane). 35. The method of any one of embodiments 30-32, wherein the nucleic acid delivery composition is a composition of any of the embodiments 1-29. 36. A method for delivery of a nucleic acid into cells comprising the step of: contacting the lipid nanoparticle (LNP) with the cells, wherein the cells are human dendritic cells, wherein the 134 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 LNP composition was obtained by a process comprising one or more steps of any one of embodiments 30-34. 37. The method of embodiment 36, wherein the LNP comprises an ionizable cationic lipid of any one of embodiments 1-29. 38. The method of any one of embodiment 36 or 37, wherein the cells are in a human or animal subject. 39. The method of embodiment 38, wherein the LNP is administered to the subject intramuscularly, subcutaneously, intradermally, or topically. 40. An LNP composition comprising: a. a nucleic acid, wherein the nucleic acid is an mRNA; b. a cholesterol sterol in a total amount of 25-40 mol% of the total lipid content of the LNP composition; c. an ionizable cationic lipid having a N / P ratio of 3 to 8 relative to the nucleic acid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; d. one or more phospholipids in a total amount of phospholipids of 5-25 mol% of the total lipid content of the LNP composition, the one or more phospholipids are selected from the group consisting of: i. an ammonium salt of dipalmitoylphosphatidyl-L-serine ((L-serine)DPPS) lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition; and ii. a distearoylphosphatidylcholine (DSPC) phospholipid in a total amount of 5-25 mol% of the total lipid content of the LNP composition; and e. a PEG-containing conjugated lipid in a total amount of 0.5 – 2.5 mol% of the total lipid content of the LNP composition. 41. A method for administering a nucleic acid to a subject in need thereof comprising administering a nucleic acid lipid nanoparticle (LNP) composition to a human subject, the nucleic acid LNP composition comprising: a. a nucleic acid; b. an ionizable cationic lipid at a N / P ratio of 3 to 8 relative to the nucleic acid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; c. a sterol in a total amount of 25-45 mol% of the total lipid content of the LNP composition; 135 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 d. one or more phospholipids in a total amount of phospholipids of 5-25 mol% of the total lipid content of the LNP composition, and comprising a phosphatidylglycerol (PG) in a total amount of 1.0 – 10 mol% of the total lipid content of the LNP composition; and e. a conjugated lipid in a total amount of 0.5 – 2.5 mol% of the total lipid content of the LNP composition. 42. A nucleic acid lipid nanoparticle (LNP) composition comprising: a. a nucleic acid; b. an ionizable cationic lipid at a N / P ratio of 3 to 8 relative to the nucleic acid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; c. a sterol in a total amount of 25-45 mol% of the total lipid content of the LNP composition; d. one or more phospholipids in a total amount of phospholipids of 5-25 mol% of the total lipid content of the LNP composition, and comprising a phosphatidylserine (PS) lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition; and e. a conjugated lipid in a total amount of 0.5 – 2.5 mol% of the total lipid content of the LNP composition. 43. The composition of embodiment 42, wherein a. the nucleic acid is mRNA; b. the ionizable cationic lipid is present in the LNP composition at a N / P ratio of 4 to 7 relative to the nucleic acid; c. the sterol is cholesterol; and d. the conjugated lipid is a PEG-containing conjugated lipid. 44. The composition of embodiment 43, wherein the one or more phospholipids comprise at least two phospholipids having mismatched acyl chain lengths. 45. The composition of embodiment 44, wherein the phosphatidylserine (PS) lipid is dipalmitoylphosphatidyl-L-serine ((L-serine)DPPS). 46. The composition of embodiment 45, wherein the one or more phospholipids comprise a phospholipid selected from the group consisting of: distearoylphosphatidylcholine (DSPC) and hydrogenated soy phosphatidylcholine (HSPC). 47. The composition of embodiment 46, wherein the one or more phospholipids consist of distearoylphosphatidylcholine (DSPC) and dipalmitoylphosphatidyl-L-serine ((L- serine)DPPS). 136 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 48. The composition of embodiment 47, wherein the PEG-containing conjugated lipid is PEG(2000)-dimyristoylglycerol (PEG-DMG). 49. An LNP composition wherein the ionizable lipid has the chemical structure: , 1, 2, 3 or 4; R2and R3are each independently (C1-C4) alkyl optionally substituted with hydroxyl; and n is an integer equal to 2, 3 or 4. 50. The composition of embodiment 49, wherein a. R2and R3are each methyl; and b. n is 3 or 4. 51. The composition of any one of embodiment 49, wherein the ionizable cationic lipid is one or more compounds selected from the group consisting of: KC3-OA, KC3-PA, KC3-C17 (8:1), and KC3-C15 (C8:1). 52. The composition of any one of embodiments 49-51, wherein the ionizable cationic lipid is KC3-PA. 53. The composition of any one of embodiments 49-51, wherein the ionizable cationic lipid is KC3-OA. 54. The composition of any one of embodiments 49-51, wherein the ionizable cationic lipid is KC3-C17 (C8:1). 55. An ionizable cationic lipid selected from the group consisting of KC3-PA, KC3-C17 (8:1), and KC3-C15 (C8:1). 56. A composition comprising an ionizable cationic lipid selected from one or more of the following: 137 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 a. a racemic mixture of 3-rac-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N- dimethylpropan-1-amine (KC3-OA racemate), or KC3-OA enantiomer; and b. 4-rac-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N-dimethylbutan-1-amine (AKG-KC4-OA racemate). 57. The composition of embodiment 56, wherein the ionizable cationic lipid is KC3-OA racemate. 58. The composition of embodiment 56, wherein the ionizable cationic lipid is KC3-OA(S) enantiomer. 59. The composition of embodiment 56, wherein the ionizable cationic lipid is KC3-OA enantiomer purified from AKG-KC3-OA racemate. 60. The composition of embodiment 56, wherein the ionizable cationic lipid is KC3-OA racemate. 61. The composition of any one of embodiments 56-60, wherein the composition is a nucleic acid lipid nanoparticle (LNP) composition. 62. The composition of embodiment 61, wherein the LNP composition comprises: a. a nucleic acid; b. the ionizable cationic lipid at a N / P ratio of 3 to 8 relative to the nucleic acid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; c. a sterol in a total amount of 0.5-50 mol% of the total lipid content of the LNP composition; d. one or more phospholipids in a total amount of phospholipids of 5-50 mol% of the total lipid content of the LNP composition; and e. a conjugated lipid in a total amount of 0.5-2.5 mol% of the total lipid content of the LNP composition. 63. The composition of embodiment 62, wherein a. the nucleic acid is mRNA; b. the ionizable cationic lipid is present in the LNP composition at a N / P ratio of 4 to 7 relative to the nucleic acid; c. the sterol is cholesterol; and d. the conjugated lipid is a PEG-containing conjugated lipid. 64. The composition of embodiment 63, wherein the one or more phospholipids comprise at least two phospholipids having mismatched acyl chain lengths. 138 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 65. The composition of any one of embodiments 62-64, wherein the one or more phospholipids comprise a phosphatidylserine (PS) lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition. 66. The composition of embodiment 65, wherein the phosphatidylserine (PS) lipid is dipalmitoylphosphatidyl-L-serine ((L-serine)DPPS). 67. The composition of embodiment 66, wherein the one or more phospholipids comprise a phospholipid selected from the group consisting of: distearoylphosphatidylcholine (DSPC) and hydrogenated soy phosphatidylcholine (HSPC). 68. The composition of embodiment 67 wherein the one or more phospholipids consist of distearoylphosphatidylcholine (DSPC) and dipalmitoylphosphatidyl-L-serine ((L- serine)DPPS). 69. The composition of embodiment 68, wherein the PEG-containing conjugated lipid is PEG(2000)-dimyristoylglycerol (PEG-DMG). 70. The composition of any one of embodiments 62-69, wherein the composition has 5-50 mol% total phospholipid. 71. The composition of embodiment 70, wherein the composition has 5, 10, 15, 20, 25, 30, 35, 40, 45 or 50 mol% total phospholipid. 72. The composition of any one of embodiments 62-71, wherein the LNP composition is further characterized by: a. the sterol in a total amount of 45.5 mol% of the total lipid content of the LNP composition; and b. the one or more phospholipids in a total amount of phospholipids of 5 mol% of the total lipid content of the LNP composition. 73. The composition of any one of embodiments 62-71, wherein the LNP composition is further characterized by: a. the sterol in a total amount of 40.5 mol% of the total lipid content of the LNP composition; and b. the one or more phospholipids in a total amount of phospholipids of 10 mol% of the total lipid content of the LNP composition. 74. The composition of any one of embodiments 62-71, wherein the LNP composition is further characterized by: 139 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 a. the sterol in a total amount of 35.5 mol% of the total lipid content of the LNP composition; and b. the one or more phospholipids in a total amount of phospholipids of 15 mol% of the total lipid content of the LNP composition. 75. The composition of any one of embodiments 62-71, wherein the LNP composition is further characterized by: a. the sterol in a total amount of 30.5 mol% of the total lipid content of the LNP composition; and b. the one or more phospholipids in a total amount of phospholipids of 20 mol% of the total lipid content of the LNP composition. 76. The composition of any one of embodiments 62-71, wherein the LNP composition is further characterized by: a. the sterol in a total amount of 25.5 mol% of the total lipid content of the LNP composition; and b. the one or more phospholipids in a total amount of phospholipids of 25 mol% of the total lipid content of the LNP composition. 77. The composition of any one of embodiments 62-71, wherein the LNP composition is further characterized by: a. the sterol in a total amount of 20.5 mol% of the total lipid content of the LNP composition; and b. the one or more phospholipids in a total amount of phospholipids of 30 mol% of the total lipid content of the LNP composition. 78. The composition of any one of embodiments 62-71, wherein the LNP composition is further characterized by: a. the sterol in a total amount of 15.5 mol% of the total lipid content of the LNP composition; and b. the one or more phospholipids in a total amount of phospholipids of 35 mol% of the total lipid content of the LNP composition. 79. The composition of any one of embodiments 62-71, wherein the LNP composition is further characterized by: 140 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 a. the sterol in a total amount of 10.5 mol% of the total lipid content of the LNP composition; and b. the one or more phospholipids in a total amount of phospholipids of 40 mol% of the total lipid content of the LNP composition. 80. The composition of any one of embodiments 62-71, wherein the LNP composition is further characterized by: a. the sterol in a total amount of 5.5 mol% of the total lipid content of the LNP composition; and b. the one or more phospholipids in a total amount of phospholipids of 45 mol% of the total lipid content of the LNP composition. 81. The composition of any one of embodiments 62-71, wherein the LNP composition is further characterized by: a. the sterol in a total amount of 0.5 mol% of the total lipid content of the LNP composition; and b. the one or more phospholipids in a total amount of phospholipids of 50 mol% of the total lipid content of the LNP composition. 82. The composition of any one of embodiments 61-81, wherein the sterol is cholesterol. 83. The composition of any one of embodiments 61-82, wherein the ionizable cationic lipid is KC3-OA. 84. The composition of embodiment 83, wherein the LNP composition comprises a total of 48- 54 mol% of the ionizable cationic lipid. 85. The composition of any one of embodiments 61-84, wherein the one or more phospholipids comprise a phosphatidylserine (PS) lipid. 86. The composition of embodiment 85, wherein the PS lipid is DPPS. 87. The composition of any one of embodiments 85-86, wherein the PS lipid is present in a total of 5 mol%. 88. The composition of any one of embodiments 61-88, wherein the LNP composition comprises a DSPC phospholipid. 89. The composition of embodiment 88, wherein the LNP composition comprises 7.5 mol% DSPC phospholipid. 141 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 90. The composition of embodiment 88, wherein the LNP composition comprises 8.5 mol% DSPC phospholipid. 91. The composition of embodiment 88, wherein the LNP composition comprises 10 mol% DSPC phospholipid. 92. The composition of embodiment 88, wherein the LNP composition comprises 11.5 mol% DSPC phospholipid. 93. The composition of embodiment 88, wherein the LNP composition comprises 12.5 mol% DSPC phospholipid. 94. The composition of embodiment 88, wherein the LNP composition comprises 13.5 mol% DSPC phospholipid. 95. The composition of embodiment 88, wherein the LNP composition comprises 14 mol% DSPC phospholipid. 96. The composition of embodiment 88, wherein the LNP composition comprises 15 mol% DSPC phospholipid. 97. The composition of embodiment 88, wherein the LNP composition comprises 16.5 mol% DSPC phospholipid. 98. The composition of embodiment 88, wherein the LNP composition comprises 17.5 mol% DSPC phospholipid. 99. The composition of embodiment 88, wherein the LNP composition comprises 20 mol% DSPC phospholipid. 100. The composition of embodiment 88, wherein the LNP composition comprises 7.5-20 mol% DSPC phospholipid. 101. The composition of any one of embodiments 61-67 or 70-87, wherein the LNP composition comprises a HSPC phospholipid. 102. The composition of embodiment 101, wherein the LNP composition comprises 10 mol% HSPC phospholipid. 103. The composition of any one of embodiments 61-102, wherein the LNP composition comprises the sterol cholesterol. 104. The composition of embodiment 103, wherein the LNP composition comprises 25 mol% cholesterol. 142 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 105. The composition of embodiment 103, wherein the LNP composition comprises 25.5 mol% cholesterol. 106. The composition of embodiment 103, wherein the LNP composition comprises 26 mol% cholesterol. 107. The composition of embodiment 103, wherein the LNP composition comprises 28 mol% cholesterol. 108. The composition of embodiment 103, wherein the LNP composition comprises 28.5 mol% cholesterol. 109. The composition of embodiment 103, wherein the LNP composition comprises 29 mol% cholesterol. 110. The composition of embodiment 103, wherein the LNP composition comprises 30 mol% cholesterol. 111. The composition of embodiment 103, wherein the LNP composition comprises 30.5 mol% cholesterol. 112. The composition of embodiment 103, wherein the LNP composition comprises 31 mol% cholesterol. 113. The composition of embodiment 103, wherein the LNP composition comprises 31.5 mol% cholesterol. 114. The composition of embodiment 103, wherein the LNP composition comprises 32 mol% cholesterol. 115. The composition of embodiment 103, wherein the LNP composition comprises 32.5 mol% cholesterol. 116. The composition of embodiment 103, wherein the LNP composition comprises 33 mol% cholesterol. 117. The composition of embodiment 103, wherein the LNP composition comprises 33.5 mol% cholesterol. 118. The composition of embodiment 103, wherein the LNP composition comprises 34 mol% cholesterol. 119. The composition of embodiment 103, wherein the LNP composition comprises 34.5 mol% cholesterol. 143 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 120. The composition of embodiment 103, wherein the LNP composition comprises 35 mol% cholesterol. 121. The composition of embodiment 103, wherein the LNP composition comprises 35.5 mol% cholesterol. 122. The composition of embodiment 103, wherein the LNP composition comprises 36 mol% cholesterol. 123. The composition of embodiment 103, wherein the LNP composition comprises 36.5 mol% cholesterol. 124. The composition of embodiment 103, wherein the LNP composition comprises 37 mol% cholesterol. 125. The composition of embodiment 103, wherein the LNP composition comprises 37.5 mol% cholesterol. 126. The composition of embodiment 103, wherein the LNP composition comprises 38 mol% cholesterol. 127. The composition of embodiment 103, wherein the LNP composition comprises 25-38 mol% cholesterol. 128. The composition of embodiment 103, wherein the LNP composition comprises 25-39 mol% cholesterol. 129. The composition of embodiment 103, wherein the LNP composition comprises 25-40 mol% cholesterol. 130. The composition of any one of embodiments 61-102, wherein the LNP composition comprises the sterol beta-sitosterol. 131. The composition of embodiment 130, wherein the LNP composition comprises 33 mol% beta-sitosterol. 132. The composition of embodiment 130, wherein the LNP composition comprises 35.5 mol% beta-sitosterol. 133. The composition of embodiment 130, wherein the LNP composition comprises 33-35.5 mol% beta-sitosterol. 134. The composition of embodiment 130, wherein the LNP composition comprises 33-35.5 mol% beta-sitosterol. 144 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 135. The composition of embodiment 130, wherein the LNP composition comprises 25-38 mol% beta-sitosterol. 136. The composition of any one of embodiments 62-135, wherein the LNP composition comprises 1.5 mol% of the PEG-containing conjugated lipid. 137. The composition of any one of embodiments 63-136, wherein the PEG-containing conjugated lipid is PEG-DMG. 138. The composition of embodiment 137, wherein the composition comprises 1.5 mol% of the PEG-DMG. 139. The composition of embodiment 137, wherein the composition comprises 2.0 mol% of the PEG-DMG. 140. The composition of embodiment 137, wherein the composition comprises 2.5 mol% of the PEG-DMG. 141. The composition of embodiment 137, wherein the composition comprises 3.0 mol% of the PEG-DMG. 142. The composition of embodiment 137, wherein the composition comprises 3.5 mol% of the PEG-DMG. 143. The composition of embodiment 137, wherein the composition comprises 4.0 mol% of the PEG-DMG. 144. The composition of any one of embodiments 63-136, wherein the PEG-containing conjugated lipid is PEG-DLG. 145. The composition of embodiment 137, wherein the composition comprises 1.0 mol% of the PEG-DLG. 146. The composition of embodiment 137, wherein the composition comprises 1.5 mol% of the PEG-DLG. 147. The composition of embodiment 137, wherein the composition comprises 2.0 mol% of the PEG-DLG. 148. The composition of embodiment 137, wherein the composition comprises 2.5 mol% of the PEG-DLG. 149. The composition of embodiment 137, wherein the composition comprises 3.0 mol% of the PEG-DLG. 145 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 150. The composition of embodiment 137, wherein the composition comprises 3.5 mol% of the PEG-DLG. 151. The composition of embodiment 137, wherein the composition comprises 4.0 mol% of the PEG-DLG. 152. The composition of embodiment 137, wherein the composition comprises 1.0-4.0 mol% of the PEG-DLG. 153. A composition selected from Formulations 1-44 in Table 87A. 154. A composition selected from Formulations 45-78 in Table 87B. 155. A composition selected from Formulations 79-116 in Table 87C. 156. A composition selected from Formulations 117-154 in Table 87D. EXAMPLES While this disclosure has been described in relation to certain embodiments, and many details have been set forth for purposes of illustration, it will be apparent to those skilled in the art that this disclosure includes additional embodiments, and that some of the details described herein may be varied considerably without departing from this disclosure. This disclosure includes such additional embodiments, modifications and equivalents. In particular, this disclosure includes any combination of the features, terms, or elements of the various illustrative components and examples. Unless explicitly indicated otherwise, the isomer form of the phosphatidylserine lipids used in the Examples is phosphatidyl-L-serine. Certain examples are provided below to illustrate various embodiments of the embodiments disclosed herein. One of ordinary skill in the art will recognize that the various embodiments disclosed herein are not limited to these specific illustrative examples. Example 1A: Synthesis of Ionizable Lipids Scheme 1 Synthesis of acid intermediates for AKG-UO-1 to AKG-UO-3. See FIGs.47A-47B. 146 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 The acid intermediates (6Z,12Z)-6,12-octadecadienoic acid and (6Z,12Z)-6,12- hexadecadienoic acid were prepared by a general synthesis, involving i) an initial Witting reaction of triphenyl phosphonium ylide, prepared from 5-bromo pentanol, and the corresponding aldehyde, ii) conversion of the terminal alcohol to bromide by mesylation and substitution, iii) repeating the sequence of ylide synthesis and Witting reaction, and finally iv) periodic acid oxidation of the terminal alcohol. The resulting acid intermediates were utilized in the synthesis of AKG-UO-1 to AKG-UO-4, vide infra. 147 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 Scheme 2 Synthesis of acid intermediate for AKG-UO-5 - UO-5 was prepared by a general synthesis shown in Scheme 2, involving i) alkylation of silyl protected 10-hydroxy-1-decyne with (5Z)-1-bromo-5-octene, ii) catalytic hydrogenation of the alkyne to a cis-alkene, iii) removal of silyl protection on the alcohol, and finally iv) oxidation of the terminal alcohol to the desired acid. Scheme 3 Synthesis of acid intermediates for AKG-BDG-01 and AKG-BDG-02 148 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 Synthesis of two disulfide acid intermediates used in the synthesis of AKG-BDG-1 and AKG- BDG-2 is shown in Scheme 4. A general synthesis of acid intermediate for AKG-BDG-1 involves i) synthesis of 4- mercapto butyric acid from 4-bromo butyric acid, ii) reaction of 4-mercapto butyric acid with DPS resulting in 4-(2-pyridinyldisulfanyl)butanoic acid iii) catalytic hydrogenation of 3-decyn-1-ol to a cis-alkene, iv) tosylation of the primary alcohol, v) displacement of the tosyl group using thiourea resulting in a terminal thiol, and finally vi) coupling of the terminal thiol with 4-(2- pyridinyldisulfanyl)butanoic acid prepared in step ii above, resulting in the disulfide containing acid intermediate. Following a similar synthetic sequence starting from 3-dodecyn-1-ol yielded the second acid intermediate used in the synthesis of AKG-BDG-2. Scheme 4 Synthesis of AKG-UO-1, AKG-UO-4, AKG-UO-5, AKG-BDG-1 and AKG-BDG-2 149 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 A general synthesis of lipids AKG-UO-1, AKG-UO-4, AKG-UO-5, AKG-BDG-1 and AKG-BDG-2 shown in Scheme 4, involves the following steps: i) tosylation of the primary alcohol of the commercially available chiral dioxolane ii) displacement of the tosyl group using dimethylamine resulting in a tertiary amine, iii) acid catalyzed deprotection of the diol, and finally iv) esterification of the diol with the corresponding acid intermediates synthesized according to Schemes 1-3. AKG-UO-2 is prepared following a similar synthetic sequence starting from a different dioxolane and a corresponding acid intermediate, as shown in Scheme 5 below. Scheme 5 Synthesis of AKG-UO-2 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 A general synthesis of trialkyl phosphate containing lipid AKG-UO-3 shown in Scheme 6, involves the following steps: i) reaction of primary alcohol of a commercially available chiral dioxolane with methyl dichlorophosphite resulting in the corresponding dialkyl chlorophosphite ii) displacement of the chloride in dialkyl chlorophosphite by treating it with 3-bromo propanol, resulting in the corresponding trialkyl phosphite iii) acid catalyzed deprotection of the diol iv) esterification of the diol with the corresponding acid intermediate synthesized according to Scheme 1, and finally v) displacement of the bromide group using dimethylamine resulting in a tertiary amine. Scheme 6 Synthesis of AKG-UO-3 Alternatively, acid intermediates having two methylene groups between double bond positions in the hydrocarbon chain are synthesized as described in Caballeira et al., Chem. Phys. Lipids, vol. 100, p. 33-40, 1999, or as described by D’yakonov et al. (D’yakonov et al., Med. Chem. Res., 2016, vol.25, p.30-39; D’yakonov et al., Chem. Commun. 2013, vol.49, p 8401- 8403; D’yakonov et al., 2020, Phytochem. Rev.). Example 1B. Synthesis of Ionizable Lipids- see FIG.48 151 ACTIVE 704513519v1 Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 1. 2-((S)-2,2-di((6Z,12Z)-octadeca-6,12-dien-1-yl)-1,3-dioxolan-4-yl)-N,N-dimethylethan-1- amine (AKG-KC2-01, O-12095) 2. 3-((S)-2,2-di((6Z,12Z)-octadeca-6,12-dien-1-yl)-1,3-dioxolan-4-yl)-N,N-dimethylpropan- 1-amine (AKG-KC3-01, O-12096) 3. 2-((S)-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N-dimethylethan-1-amine (AKG-KC2-OA, O-11880) 4. 2-((S)-2,2-di((Z)-hexadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N-dimethylethan-1-amine (AKG-KC2-PA, O-11879) 5. 3-((S)-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-y...

Claims

Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 CLAIMS What is claimed is:

1. A lipid nanoparticle (LNP) composition comprising an ionizable cationic lipid selected from one or more of the following: a. a mixture of 3-(S)-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N- dimethylpropan-1-amine (KC3-OA(S)) and 3-(R)-2,2-di((Z)-octadec-9-en-1-yl)-1,3- dioxolan-4-yl)-N,N-dimethylpropan-1-amine (KC3-OA(R)) enantiomer; and b. 4-(S)-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N-dimethylbutan-1-amine (KC4-OA(S)) and 4-(R)-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N- dimethylbutan-1-amine (KC4-OA(R)).

2. The LNP composition of claim 1, wherein the ionizable cationic lipid is a. a racemic mixture of 3-rac-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N- dimethylpropan-1-amine (KC3-OA racemate); or b. a racemic mixture of 4-rac-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N- dimethylbutan-1-amine (KC4-OA racemate).

3. The LNP composition of claim 1, wherein the ionizable cationic lipid is a. 3-(R)-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N-dimethylpropan-1-amine (KC3-OA (R)); or b. 4-(R)-2,2-di((Z)-octadec-9-en-1-yl)-1,3-dioxolan-4-yl)-N,N-dimethylbutan-1-amine (KC4-OA (R)).

4. The LNP composition of any one of claims 1-3, wherein the LNP composition comprises: a. a nucleic acid; b. the ionizable cationic lipid at a N / P ratio of 3 to 8 relative to the nucleic acid in a total amount of 40-65 mol% of the total lipid content of the LNP composition; c. a sterol in a total amount of 0.5-50 mol% of the total lipid content of the LNP composition; d. one or more phospholipids in a total amount of phospholipids of 5-50 mol% of the total lipid content of the LNP composition; and e. a conjugated lipid in a total amount of 0.5-2.5 mol% of the total lipid content of the LNP composition.

5. The LNP composition of claim 4, wherein 290 ACTIVE 704513519v1Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 a. the nucleic acid is mRNA; and b. the ionizable cationic lipid is present in the LNP composition at a N / P ratio of 4 to 7 relative to the nucleic acid.

6. The LNP composition of claim 5, wherein the one or more phospholipids comprise a phosphatidylserine (PS) lipid in a total amount of 2.5-10 mol% of the total lipid content of the LNP composition.

7. The LNP composition of claim 6, wherein the phosphatidylserine (PS) lipid is dipalmitoylphosphatidyl-L-serine ((L-serine)DPPS).

8. The LNP composition of claim 7, wherein the one or more phospholipids comprise at least two phospholipids having mismatched acyl chain lengths.

9. The LNP composition of claim 8, wherein the one or more phospholipids comprise a phospholipid selected from the group consisting of: distearoylphosphatidylcholine (DSPC), hydrogenated soy phosphatidylcholine (HSPC), and dipalmitoylphosphatidyl-L-serine ((L- serine)DPPS).

10. The LNP composition of claim 9, wherein the sterol is cholesterol or beta-sitosterol.

11. The LNP composition of claim 10, wherein the conjugated lipid is selected from the group consisting of: PEG(2000)-dimyristoylglycerol (PEG-DMG), 1,2-dilauroyl-sn-glycero-3- phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (PEG-DLPE), and PEG(2000)-dilauroylglycerol (PEG-DLG).

12. The LNP composition of claim 11, wherein the LNP composition comprises 25-40 mol% cholesterol.

13. The LNP composition of claim 12, wherein the LNP composition comprises 1.5-4.0 mol% of the conjugated lipid.

14. The LNP composition of claim 13, wherein the one or more phospholipids comprise distearoylphosphatidylcholine (DSPC).

15. The LNP composition of claim 14, wherein the PEG-containing conjugated lipid is PEG(2000)-dimyristoylglycerol (PEG-DMG) or PEG(2000)-dilauroylglycerol (PEG-DLG).

16. The LNP composition of claim 1, wherein the ionizable cationic lipid is a KC3-OA racemate of KC3-OA(S) and KC3-OA(R).

17. The LNP composition of claim 1, wherein the ionizable cationic lipid is KC3-OA(R) enantiomer. 291 ACTIVE 704513519v1Attorney Docket No.191016-010901 / PCT Electronically Filed: November 27, 2024 18. The LNP composition of claim 1, wherein the ionizable cationic lipid is a KC3-OA enantiomer purified from KC3-OA racemate of KC3-OA(S) and KC3-OA(R).

19. The LNP composition of claim 1, wherein the ionizable cationic lipid is KC3-OA mixed enantiomers of KC3-OA(S) and KC3-OA(R), or KC4-OA mixed enantiomers of KC4-OA(S) and KC4-OA(R).

20. A vaccine composition comprising the LNP composition of any one of claims 1-19. 292 ACTIVE 704513519v1

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