Ionizable lipid compounds comprising amidine crosslinkers or aryl-alkyl disulfides and lipid nanoparticles (LNPS) comprising the same

Abstract

The disclosure relates, in one aspect, to ionizable lipid compounds of formula (I) comprising an aryl-alkyl disulfide moiety. The disclosure relates, in another aspect, to ionizable lipid compounds of formula (II) comprising an amidine moiety. In another aspect, the disclosure relates to lipid nanoparticles (LNPs) comprising at least one ionizable lipid of formula (I) or (II). In another aspect, the disclosure provides methods of the LNPs of the disclosure for delivery of therapeutic cargo.

Description

[0001]Attorney Docket No.0466483-7483WO1 TITLE OF THE INVENTION Ionizable Lipid Compounds Comprising Amidine Crosslinkers or Aryl-Alkyl Disulfides and Lipid Nanoparticles (LNPs) Comprising the Same CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority under 35 U.S.C. § 119(e) to U.S. Provisional Patent Application No.63 / 709,250, filed October 18, 2024, and U.S. Provisional Patent Application No.63 / 710,116, filed October 22, 2024, all of which applications are incorporated herein by reference in their entireties. STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT This invention was made with government support under TR002776 awarded by the National Institutes of Health. The government has certain rights in the invention. REFERENCE TO AN ELECTRONIC SEQUENCE LISTING The XML file named “046483-7483WO1 - Sequence Listing.xml” created on October 16, 2025, comprising 2,087 bytes, is incorporated herein by reference in its entirety. BACKGROUND Messenger RNA (mRNA)-based therapeutics have revolutionized treatments across a wide range of applications for vaccination, protein replacement therapy, cancer immunotherapy, and gene editing. Among non-viral delivery platforms, lipid nanoparticles (LNPs), comprised of an ionizable lipid, phospholipid, cholesterol, and poly(ethylene glycol) (PEG) lipid, have emerged as the most promising platforms for mRNA therapeutic delivery. Despite advancements in LNP technoloogy, the discontinuous vasculature of hepatic sinusoids results in preferential accumulation of mRNA-LNPs in the liver, confining transgene expression exclusively to hepatic cells following systemic administration. Thus, there is a need in the art for ionizable lipids and lipid nanoparticles comprising the same which permit targeting of extrahepatic tissues (e.g., lymph nodes) and / or dendritic cells for enhanced mRNA therapeutic delivery. The present disclosure addresses this need. BRIEF SUMMARY OF THE INVENTION - 1 - 53224792.3 Attorney Docket No.0466483-7483WO1 In one aspect, the disclosure provides a compound of formula (I), or a salt, stereoisomer, or isotopologue thereof: (I), wherein: A is selected from the group consisting of ; each occurrence of L1, if from the group consisting of -(optionally substituted C1- -, - C2-C12alkenylenyl)-, - (optionally substituted C1-C12 alkynylenyl)-, -(optionally substituted C1-C12 heteroalkylenyl)- , -(optionally substituted C3-C8cycloalkylenyl)-, -(optionally substituted C2-C8heterocyloalkylenyl)-, -(optionally substituted C6-C10 arylenyl)-, -(optionally substituted C2- C8heteroarylenyl)-, -(optionally substituted C1-C12alkylenyl)-X-, -(optionally substituted C2- C12 alkenylenyl)-X-, -(optionally substituted C1-C12 alkynylenyl)-X-, -(optionally substituted C1-C12heteroalkylenyl)-X-, -(optionally substituted C3-C8cycloalkylenyl)-X-, -(optionally substituted C2-C8 heterocyloalkylenyl)-X-, -(optionally substituted C6-C10 arylenyl)-X-, and - (optionally substituted C2-C8heteroarylenyl)-X-; each occurrence of X, if present, is independently selected from the group consisting of -N(R1e)-, -N(RA)-, and -O-; each occurrence of R1a, R1b, R1c, R1d, and R1e, if present, is independently , wherein: R1cand R1dcan combine with the atoms to which they are bound to form an optionally substituted C2-C8 heterocycloalkyl, or one of R1cand R1dcan combine with one occurrence of L1to form an optionally substituted C2-C8 heterocycloalkyl; R2is selected from the group consisting of optionally substituted C1-C6alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C6 heteroalkyl, optionally substituted C3-C6 heteroalkenyl, optionally substituted C2-C8 heterocycloalkyl, and N(RA)(RB); each occurrence of L2is independently selected from the group consisting of - (optionally substituted C1-C6alkylenyl)-, -C(=O)-, -O-, and -N(RA)-; each occurrence of R3is independently selected from the group consisting of - 2 - 53224792.3 Attorney Docket No.0466483-7483WO1 optionally substituted C1-C24 alkyl and optionally substituted C1-C24 heteroalkyl; each occurrence of Ar is independently selected from the group consisting of - (optionally substituted C6-C10 heteroarylenyl)- and -(optionally substituted C2-C8 heteroarylenyl)-; m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; and each occurrence of RAand RBis independently selected from the group consisting of H, optionally substituted C1-C6alkyl, optionally substituted C2-C6alkenyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C6 heteroalkyl, optionally substituted C3-C6heteroalkenyl, optionally substituted C2-C8heterocycloalkyl, optionally substituted C6- C10 aryl, and optionally substituted C2-C8 heteroaryl. In certain embodiments, the compound of formula (I) is selected from the group consisting of: R1a (a) at least one ionizable lipid comprising the compound of formula (I); (b) at least one neutral lipid; (c) at least one cholesterol lipid and / or a modified derivative thereof; and (d) at least one polymer-conjugated lipid and / or a modified derivative thereof. In another aspect, the disclosure provides a pharmaceutical composition comprising the lipid nanoparticle (LNP) comprising the ionizable lipid compound of formula (I) and at least one pharmaceutically acceptable carrier. In another aspect, the disclosure provides a method of treating, preventing, and / or ameliorating a disease or infection in a subject. In certain embodiments, the method comprises administering to the subject at least one lipid nanoparticle (LNP) comprising the ionizable lipid compound of formula (I) or a pharmaceutical composition thereof. In another aspect, the disclosure provides a method of delivering at least one therapeutic cargo molecule to a lymph node of a subject. In certain embodiments, the method - 3 - 53224792.3 Attorney Docket No.0466483-7483WO1 comprises administering to the subject at least one lipid nanoparticle (LNP) comprising the ionizable lipid compound of formula (I) or a pharmaceutical composition thereof. In another aspect, the disclosure provides a compound of formula (II), or a salt, stereoisomer, or isotopologue thereof: , wherein: R1a, R1b, and a polyamine moiety, wherein each amine substituent of the polyamine moiety is substituted with at least one optionally substituted C1-C20alkyl, optionally substituted C1-C20heteroalkyl, optionally substituted C2-C20 alkenyl, optionally substituted C2-C20 heteroalkyenyl, optionally substituted C2-C20alkynyl, and optionally substituted C2-C20heteroalkynyl; R2a, R2b, and each occurrence of R2c, if present, are each independently selected from the and optionally substituted C1-C6 alkyl; and each occurrence of R3c, if present, are each independently selected from the and optionally substituted C1-C6 alkyl; each occurrence of R4aand R4bis independently selected from the group consisting of H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C8cycloalkyl, optionally substituted C2-C8heterocycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted C2-C8 heteroaryl; each occurrence of R5aand R5bis independently selected from the group consisting of H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C8cycloalkyl, optionally substituted C2-C8heterocycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted C2-C8 heteroaryl; each occurrence of R6is ; each occurrence of R7aand R7b, if selected from the group consisting of H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C6-C10aryl, and optionally substituted C2-C8heteroaryl; - 4 - 53224792.3 Attorney Docket No.0466483-7483WO1 m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each occurrence of n is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; o is 0, 1, 2, 3, 4, or 5; and each occurrence of p is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In certain embodiments, the compound of formula (II) is selected from the group consisting of: (a) at least one ionizable lipid comprising the compound of formula (II); (b) at least one neutral lipid; (c) at least one cholesterol lipid and / or a modified derivative thereof; and (d) at least one polymer-conjugated lipid and / or a modified derivative thereof. In another aspect, the disclosure provides a pharmaceutical composition comprising the lipid nanoparticle (LNP) comprising the ionizable lipid of formula (II) and at least one pharmaceutically acceptable carrier. In another aspect, the disclosure provides a method of treating, preventing, and / or ameliorating a disease in a subject. In certain embodiments, the method comprises administering to the subject at least one lipid nanoparticle (LNP) comprising the ionizable lipid compound of formula (II) or a pharmaceutical composition thereof. In another aspect, the disclosure provides a method of delivering at least one therapeutic cargo molecule to a dendritic cell of a subject. In certain embodiments, the method comprises administering to the subject at least one lipid nanoparticle (LNP) comprising the ionizable lipid compound of formula (II) or a pharmaceutical composition thereof. - 5 - 53224792.3 Attorney Docket No.0466483-7483WO1 BRIEF DESCRIPTION OF THE FIGURES The drawings illustrate generally, by way of example, but not by way of limitation, various embodiments of the present application. FIGs.1A-1D: Certain characterization data, including encapsulation efficiency (FIG. 1A), charge (FIG.1B), hydrodynamic diameter (FIG.1C), and polydispersity index (FIG. 1D), for exemplary LNPs comprising certain aromatic ionizable lipids of the disclosure prepared using amine core 383 (e.g., C6oPhE-383, C6mPhE-383, and C6pPhE-383, inter alia) and certain controls and / or comparators (i.e., C12-200, SM-102, C6-383, C8-383, C10- 383, and C12-383, inter alia). FIGs.2A-2B: Certain in vivo characterization data, including transfection efficacy (FIG.2A) and relative toxicity (FIG.2B), for exemplary LNPs of the disclosure, utilizing HepG2 cells (20,000 cells / well, 30 ng / well luciferase mRNA). FIGs.3A-3C: Delivery and transfection of mRNA cargo utilizing certain exemplary LNPs of the disclosure in the liver by intravenous administration (FIG.3A) or intramuscular administration (FIG.3B) as compared to distribution to the full body (FIG.3C). Intramuscular administration (1 µg mRNA / mouse), intravenous administration (2 µg mRNA / mouse), imaged 6 h post administration. FIGs.4A-4C: Transfection of the inguinal lymph node (FIG.4A), iliac lymph node (FIG.4B), or both (FIG.4C) of mice intramuscularly administered (1 µg mRNA / mouse) certain exemplary LNPs of the disclosure comprising mRNA cargo 6 h post administration. FIGs.5A-5C: Comparison of delivery and transfection of mRNA cargo in the inguinal lymph node (FIG.5A), iliac lymph node (FIG.5B), or both (FIG.5C) to that of the liver in mice intramuscularly administered (1 µg mRNA / mouse) certain exemplary LNPs of the disclosure comprising mRNA cargo 6 h post administration. FIGs.6A-6C: Delivery and transfection of mRNA cargo utilizing certain exemplary LNPs of the disclosure in the liver (FIGs.6A-6B) as compared to distribution to the full body (FIG.6C). Intramuscular administration (1 µg mRNA / mouse), imaged 6 h post administration. FIG.7: scheme depicting a non-limiting, exemplary dosing scheme of the disclosure, indicating priming at day 0 (D0), a boost at day 21 (D21), and sacrifice (D40), wherein samples are collecting at multiple time points throughout. C57BL / 6J mice were administered 2 different doses (i.e., 1 µg or 5 µg COVID receptor-binding domain (RBD) mRNA); 4 mice per group; wherein groups were administered PBS (control), or LNPs comprising SM-102 - 6 - 53224792.3 Attorney Docket No.0466483-7483WO1 (comparator), C8mPhE-383, or C8pPhE-383. FIG.8: Bar graphs depicting COVID RBD IgG antibody titers observed in C57BL / 6J mice 3 weeks post-boost, wherein mice were treated in accordance with the dosing regimen described elsewhere herein (see FIG.7). FIGs.9A-9D: provides graphs depicting mouse aspartate aminotransferase (FIG.9A), mouse weight (FIG.9B), mouse alanine aminotransferase (FIG.9C), and COVID RBD antibody titers, showing that no substantial toxicity was observed C57BL / 6J mice with administration of certain LNPs of the disclosure comprising mRNA cargo. FIG.10: Exemplary modular components of certain aryl-alkyl disulfide compounds of the disclosure and corresponding numerical identifiers. FIGs.11A-11B: Exemplary synthetic scheme (FIG.11A) for the preparation of exemplary aryl-alkyl disulfide ionizable lipids of the disclosure, and exemplary aryl-alkyl disulfide ionizable lipid compounds of the disclosure. FIGs.12A-12D: Organ specific mRNA delivery of certain exemplary LNPs of the disclosure following intramuscular injection. FIG.12A: Liver transfection. FIG.12B: Lymph node transfection. FIG.12C: Total flux of lymph nodes:liver. FIG.12D: imaging of exemplary organs (e.g., heart, lungs, liver, kidney, spleen, and lymph nodes) following intramuscular administration of exemplary LNPs of the disclosure. FIGs.13A-13D: Design and evaluation of imidoester crosslinker-based ionizable lipid library for dendritic cell mRNA transfection. FIG.13A: Synthetic scheme for preparation of imidoester crosslinker-based ionizable lipids. Three different types of imidoester crosslinkers were reacted with amine cores, followed by lipid tail conjugation via an epoxide ring opening reaction. FIG.13B: Chemical structures of certain exemplary linear and cyclic amine cores used to generate the ionizable lipid library. FIG.13C: Molar ratio of each lipid component for certain exemplary LNP formulations of the disclosure and schematic illustration of LNP formulation for mRNA delivery. FIG.13D: In vitro mRNA expression screening results of the ionizable lipid library shown in heatmaps; dots indicate the amine core and shaded regions indicate the crosslinkers. BMDCs were treated with luciferase mRNA-loaded LNPs for 6 h, and luminescence intensity was represented as a heatmap according to the type of the ionizable lipids. FIGs.14A-14F: In vitro mRNA expression screening of the ionizable lipid library. FIG.2A: Structures of 2a-, 3a-, and 4-a crosslinkers were characterized by1H-NMR. FIG. 14B: In vitro mRNA expression screening result of the ionizable lipid library represented as a bar graph. FIG.14C: Comparison between the ionizable lipid without a linker and those with - 7 - 53224792.3 Attorney Docket No.0466483-7483WO1 2a linkers according to amine compounds that showed increased mRNA expression. FIG. 14D: mRNA expression of amine J- and amine N-based ionizable lipids. FIG.14E: Comparison between the ionizable lipid without a linker and those with 2a linkers according to amine compounds that showed decreased mRNA expression. FIG.14F: mRNA expression of amine I- and amine M- based ionizable lipids. Data are presented as mean ± SD (n=8 or n=5 biologically independent samples). Statistical differences were analyzed by one-way ANOVA with Tukey’s post hoc test. FIGs.15A-15D: Structural simulation and evaluation of amine N-based ionizable lipids. FIG.15A: Structure of amine N and schematic illustration of its ionizable lipids. Dots indicate the amine core and shaded regions indicate the crosslinkers. FIG.15B: Structures of amine core N-based ionizable lipids and their molecular electrostatic potentials (MEPs). Amine cores are labeled and conformation angles and distances between lipid-conjugated ionizable amines are indicated. FIG.15C: Simulated structures of C12-N and C12-2aN lipids with excipient lipids on the LNP surface. FIG.15D: Endosomal escape and reporter mRNA expression of each ionizable lipids. For endosomal escape, BMDCs were treated with Cy5- labeled mRNA-loaded LNPs, and lysosomes were stained with Lysotracker at 6 h. For reporter mRNA expression, BMDCs were treated with GFP mRNA-loaded LNPs, and the fluorescence was measured at 24 h. FIGs.16A-16D: Energy value of structural simulation of amine N-based ionizable lipids and evaluation of their antigen presentation. FIG.16A: Simulated structures of C12- 3aN and C12-4aN lipids with excipient lipids on the LNP surface. FIG.16B: Thermodynamic energy of ionizable lipids, DOPE and C14-PEG in existing individually and interacting each other. FIGs.16C-16D: Antigen presentation induced LNP treatments. BMDCs were treated with mOVA-loaded LNPs, and the antigen presentation was evaluated after 24 h incubation. The population of cells showing antigen presentation was analyzed by flow cytometry (FIG. 16C) and the quantified cell population was plotted (FIG.16D). Data are presented as mean ± SD (n=5 biologically independent samples). Statistical differences were analyzed by one-way ANOVA with Tukey’s post hoc test. FIGs.17A-17K: Characterization and intramuscular administration of C12-2aNP. FIG.17A: Chemical structure of C12-N and C12-2aN. Piperazine amine cores are indicated by a circle and the crosslinker is indicated by a box. FIG.17B: Zeta potential of C12-NP and C12-2aNP. FIG.17C: Determined pKa of C12-NP and C12-2aNP. FIG.17D: Cryo-TEM images of C12-NP and C12-2aNP. FIG.17E: Particle size of C12-NP and C12-2aNP. FIG. 17F: Polydispersity index (PDI) of the particle size of C12-NP and C12-2aNP. FIG.17G: - 8 - 53224792.3 Attorney Docket No.0466483-7483WO1 mRNA encapsulation efficiency of C12-NP and C12-2aNP. FIG.17H: Cell viability of C12- NP and C12-2aNP. FIGs.17I-17K: In vivo luminescence imaging of mice treated with luciferase mRNA-loaded LNPs. C12-NP or C12-2aNP was administered by intramuscular injection, and the mice were imaged at 6, 24, 48, and 72 h. The representative whole-body image at 6 h (FIG.17I) is shown. Luminescence intensity at the injection site at each time point (FIG.17J) and AUC (FIG.17K) are shown. Data are presented as mean ± SD (n=5 biologically independent samples). Statistical differences were analyzed by one-way ANOVA with Tukey’s post hoc test. FIGs.18A-18L: C12-2aNP-mediated metabolic reprogramming of BMDC. FIGs. 18A-18B: Acidification of cell media upon LNP treatment to BMDC. The cell media 24 h after LNP treatment (FIG.18A) was shown, and the pH of the media (FIG.18B) was measured. FIG.18C: Expression and phosphorylation of Akt and S6K, showing mTORC2 and mTORC1 activity, were evaluated by western blot. FIG.18D: The relative ratio of phosphorylated Akt to total Akt was calculated. FIGs.18E-18G: Transcriptome analysis of BMDC upon LNP treatment. Total mRNA expression was analyzed by mRNA-Seq, and gene expression level related to glycolysis were shown as heatmaps (FIG.18E). Gene set enrichment analysis of the hallmark glycolysis gene set in the C12-2aNP-treated group compared to the untreated group is shown (FIG.18F). Pathway enrichment analysis of the C12-2aNP-treated group compared to the untreated (FIG.18G). FIGs.18H-18L: Glycolytic and respiratory function of BMDCs were measured by Seahorse assay, and ECAR (FIG. 18H) and OCR (FIG.18I) are shown, respectively. Basal glycolysis and glycolytic capacity (FIG.18J). The energy phenotype of each group in basal condition is shown (FIG.18K). ATP production rates were calculated (FIG.18L). Data are presented as mean ± SD (n=3, FIGs. 18A-18D; n=3 for untreated and C12-NP or 4 for C12-2aNP, FIGs.18E-18F; n=5, FIGs. 18G-18L; biologically independent samples). Statistical differences were analyzed by one- way ANOVA with Tukey’s post hoc test. FIGs.19A-19F: Structural characterization and metabolic regulation effect of C12- 2aNP. FIG.19A: Structures of C12-N, 2aN, and C12-2aN were characterized by LC-MS. FIG.19B: Unedited western blot images of p-Akt, Akt, p-S6K, S6k, and b-actin for evaluating metabolic modulation in BMDC. FIG.19C: Gene set enrichment analysis of the hallmark glycolysis in C12-2NP treated group compared to that of untreated was shown. FIG. 19D: Calculated basal respiration and maximal respiration from Mito Stress Test was plotted. FIGs.19E-19F: Acidification of cell media upon LNPs treatment to bone marrow-derived macrophage (FIG.19E) and monocyte (FIG.19F). The pH of the media was measured 24 h - 9 - 53224792.3 Attorney Docket No.0466483-7483WO1 after the LNP treatment. Data are presented as mean ± SD (n=3 or 5 biologically independent samples). Statistical differences were analyzed by one-way ANOVA with Tukey’s post hoc test. FIGs.20A-20N: Immune responses induced by spike protein RBD mRNA-loaded C12-2aNP for SARS-CoV-2 vaccination. FIG.20A: A scheme of the vaccination strategy and analysis for the SARS-CoV-2 model. Mice were intramuscularly injected with LNPs (0.1 mg / kg of SARS-CoV-2 spike protein RBD mRNA) using a prime-boost strategy with a three- week interval. Immune responses were evaluated on day 28 after the boost injection. FIG. 20B: RBD-specific IgG antibody titers were determined by ELISA. FIGs.20C-20D: RBD- specific B cells in the spleen were analyzed by flow cytometry. The quantified cell populations (FIG.20C) and flow cytometry dot plots (FIG.20D) are shown. FIGs.20E-20F: Neutralizing antibody levels were determined by a pseudovirus neutralization assay (FIG. 20E), and the 50% neutralization titer was calculated (FIG.20F). FIG.20G: RBD-specific T cells in the spleen were analyzed by ELISpot, and IFN-g secreting cells were counted. FIG. 20H: RBD-specific T cell immune responses were evaluated by intracellular cytokine analysis. FIG.20I: Representative immunofluorescence images of vaccine-induced germinal centers (GC) in the spleen. FIGs.20J-20L: Cell populations of RBD-specific GC B cells (FIG.20J), RBD-specific memory B cells (FIG.20K), and follicular helper T cells (FIG. 20L) in the spleen were analyzed by flow cytometry. FIG.20M: CD8+memory T cell subpopulations in the spleen were analyzed by flow cytometry. FIG.20N: On day 120, RBD- specific antibody-secreting cells in the bone marrow were analyzed by ELISpot. Data are presented as mean ± SD (n=5 biologically independent samples). Statistical differences were analyzed by one-way ANOVA with Tukey’s post hoc test. FIGs.21A-21E: Immunogenicity and toxicity of C12-2aNP. FIG.21A: Gene expression level related to immunogenicity were analyzed by mRNA-Seq and shown as heatmaps. FIG.21B: Serum Il-6 level was monitored for 7 days Body weight was monitored for 14 days after intracellular injection of LNPs at a dose of 2 mg of mLuc. FIG.21C: Serum Il-6 level measured on day 1 was plotted. Data are presented as mean ± SD (n=5 biologically independent samples). Statistical differences were analyzed by one-way ANOVA with Tukey’s post hoc test. FIGs.22A-22L: Immune responses induced by OVA mRNA-loaded C12-2aNP for cancer vaccination. FIG.22A: A scheme of the vaccination strategy and analysis for a preventive cancer vaccine model. Mice were intramuscularly injected with LNPs (0.1 mg / kg of OVA mRNA) using a prime-boost strategy with a five-day interval. Immune responses - 10 - 53224792.3 Attorney Docket No.0466483-7483WO1 were evaluated 2 days after the boost injection. OVA-expressing B16F10 cells (5×105cells) were subcutaneously inoculated 5 days after the boost injection and tumor growth was monitored. FIG.22B: OVA-specific IgG antibody titers were determined by ELISA. FIG. 22C: OVA-specific T cells in the spleen were analyzed by ELISpot, and IFN-g secreting cells were counted. FIGs.22D-22E: OVA-specific T cells in the spleen were analyzed by flow cytometry using an OVA-tetramer. The quantified cell populations (FIG.22D) and flow cytometry dot plots (FIG.22E) are shown. FIG.22F: Tumor growth was monitored over 23 days. FIG.22G: A scheme of the vaccination strategy and analysis for a therapeutic cancer vaccine model. OVA expressing B16F10 cells (5×105cells) were subcutaneously inoculated, and the mice were intramuscularly injected with LNPs (2 mg of OVA mRNA per injection) using a prime-boost strategy with a five-day interval after 5 days of the tumor inoculation. FIG.22H: Tumor growth was monitored over 20 days. FIG.22I: Survival rate was monitored over 44 days. FIG.22J: Representative immunofluorescence images of vaccine-induced T cell infiltration and activation in tumor tissue. FIGs.22K-22L: Tumor infiltrating CD8 T cells (FIG.22K) and IFN-g-expressing population among the CD8 T cells (FIG.22L) were analyzed by flow cytometry. Data are presented as mean ± SD (n=5 biologically independent samples). Statistical differences were analyzed by one-way ANOVA with Tukey’s post hoc test. FIGs.23A-23E: Representative flow cytometry plots and ELISpot images in the SARS-CoV-2 vaccine model. FIG.23A: Flow cytometry plots for intracellular cytokine expression of antigen specific T cells. FIG.23B: Flow cytometry plots for cell populations of follicular helper T cells in the spleen. FIG.23C: Flow cytometry plots for CD8+memory T cells in the spleen. FIG.23D: ELISpot images showing IFN-g-expressing RBD specific T cells. FIG.23E: ELISpot image showing RBD-specific antibody secreting cells in the bone marrow. FIGs.24A-24C: ELISpot images, individual tumor volume and representative flow cytometry plots in the cancer vaccine model. FIG.24A: ELISpot images showing IFN-g- expressing OVA specific T cells. FIG.24B: Tumor volumes measured for 23 days in the preventive cancer vaccine model were plotted individually. FIG.24C: Tumor volumes measured for 20 days in the therapeutic cancer vaccine model were plotted individually. FIG. 24D: Flow cytometry plots for tumor infiltrating CD8+ T cells in the tumor. FIG.24E: Flow cytometry plots for IFN-g-expressing CD8+ T cells in the tumor. DETAILED DESCRIPTION OF THE INVENTION - 11 - 53224792.3 Attorney Docket No.0466483-7483WO1 Reference will now be made in detail to certain embodiments of the disclosed subject matter, examples of which are illustrated in part in the accompanying drawings. While the disclosed subject matter will be described in conjunction with the enumerated claims, it will be understood that the exemplified subject matter is not intended to limit the claims to the disclosed subject matter. Throughout this document, values expressed in a range format should be interpreted in a flexible manner to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited. For example, a range of "about 0.1% to about 5%" or "about 0.1% to 5%" should be interpreted to include not just about 0.1% to about 5%, but also the individual values (e.g., 1%, 2%, 3%, and 4%) and the sub-ranges (e.g., 0.1% to 0.5%, 1.1% to 2.2%, 3.3% to 4.4%) within the indicated range. The statement "about X to Y" has the same meaning as "about X to about Y," unless indicated otherwise. Likewise, the statement "about X, Y, or about Z" has the same meaning as "about X, about Y, or about Z," unless indicated otherwise. In this document, the terms "a," "an," or "the" are used to include one or more than one unless the context clearly dictates otherwise. The term "or" is used to refer to a nonexclusive "or" unless otherwise indicated. The statement "at least one of A and B" or "at least one of A or B" has the same meaning as "A, B, or A and B." In addition, it is to be understood that the phraseology or terminology employed herein, and not otherwise defined, is for the purpose of description only and not of limitation. Any use of section headings is intended to aid reading of the document and is not to be interpreted as limiting; information that is relevant to a section heading may occur within or outside of that particular section. All publications, patents, and patent documents referred to in this document are incorporated by reference herein in their entirety, as though individually incorporated by reference. In the methods described herein, the acts can be carried out in any order, except when a temporal or operational sequence is explicitly recited. Furthermore, specified acts can be carried out concurrently unless explicit claim language recites that they be carried out separately. For example, a claimed act of doing X and a claimed act of doing Y can be conducted simultaneously within a single operation, and the resulting process will fall within the literal scope of the claimed process. Description: Aryl-Alkyl Disulfide Ionizable Lipid Compounds Lipid nanoparticles (LNPs) are a clinically advanced mRNA vaccine platform, as - 12 - 53224792.3 Attorney Docket No.0466483-7483WO1 demonstrated by the COVID-19 mRNA vaccines, as they can robustly transfect lymph nodes and elicit a strong immune response. However, these LNPs also highly distribute into the liver, leading to hepatic toxicity. Furthermore, lipid tail length and regiochemistry can functionally change how the lipids interact with mRNA, yet these aspects of lipid design have not yet been widely explored, and more broadly, the relationship between lipid design and LNP performance is poorly understood. In one aspect, the disclosure relates to the synthesis of a library of 14 novel ionizable lipids (ILs) with bioreducible disulfide bonds and varied tail lengths and regiochemistries. It was hypothesized that these moieties would reduce lipid toxicity while enhancing transfection, as LNPs with aromatic rings have demonstrated significantly improved transfection in vivo when compared to LNPs without aromatic rings. These ILs further enabled investigation of the structure-function relationship between IL tail length, regiochemistry, and transfection in lymph nodes and the liver. Mice were intramuscularly administered LNPs utilizing the LNPs of the disclosure and gold standards C12-200 and SM-102 and it was found that LNPs comprising ILs with a 6-10 carbon tail length in the meta and para positions outperformed C12-200 and SM-102 in achieving a high lymph node to liver delivery ratio. Further, LNPs containing ILs with disulfide bonds and aromatic rings outperformed similar ILs without these moieties in achieving high lymph node and decreased liver delivery, indicating that aromatic rings and disulfide bonds may play a central role in retaining LNPs in the endothelium. Thus, the LNPs of the disclosure, and methods of use thereof, represent an effective mRNA vaccine platform. Description: Amidine-Crosslinker Containing Ionizable Lipid Compounds In one aspect, the disclosure describes potent vaccine efficacy of the lipid nanoparticles (LNPs) of the disclosure, as demonstrated by LNP-mediated metabolic reprogramming toward glycolysis, along with efficient antigen mRNA delivery. Through imidoester crosslinker-based conjugation chemistry, a library of 80 ionizable lipids with various linked ionizable amine core structures was generated. This high-throughput synthesis of ionizable lipids with structural diversity enabled the discovery of a multifunctional ionizable lipid, C12-2aN, capable of both efficient mRNA delivery and metabolic modulation. LNPs formulated with C12-2aN not only facilitated efficient endosomal escape of mRNA for vaccine antigen expression, but also effectively stimulated glycolysis, resulting in robust vaccine efficacy in SARS-CoV-2. The synergistic effect of the LNPs induced notable neutralization of pseudovirus infection and antigen-specific memory cell - 13 - 53224792.3 Attorney Docket No.0466483-7483WO1 differentiation, contributing to long-term protection. Additionally, in ovalbumin (OVA) cancer models, vaccination with the LNPs induced infiltration of IFN-g expressing lymphocytes, leading to tumor growth inhibition and improved survival rates. Since LNPs are the most clinically advanced mRNA vaccine delivery vehicles, proven by their remarkable success as SARS-CoV-2 vaccines, the integration of mRNA delivery and metabolic reprogramming into a single LNP formulation offers significant potential as a next generation mRNA vaccine. Definitions The term “about” as used herein can allow for a degree of variability in a value or range, for example, within 10%, within 5%, or within 1% of a stated value or of a stated limit of a range, and includes the exact stated value or range. The term “alkenyl” as used herein refers to straight and branched chain and cyclic alkyl groups as defined herein, except that at least one double bond exists between two carbon atoms. Thus, alkenyl groups have from 2 to 40 carbon atoms, or 2 to about 20 carbon atoms, or 2 to 12 carbon atoms or, in some embodiments, from 2 to 8 carbon atoms. Examples include, but are not limited to vinyl, -CH=C=CCH2, -CH=CH(CH3), - CH=C(CH3)2, -C(CH3)=CH2, -C(CH3)=CH(CH3), -C(CH2CH3)=CH2, cyclohexenyl, cyclopentenyl, cyclohexadienyl, butadienyl, pentadienyl, and hexadienyl among others. The term “alkoxy” as used herein refers to an oxygen atom connected to an alkyl group, including a cycloalkyl group, as are defined herein. Examples of linear alkoxy groups include but are not limited to methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, and the like. Examples of branched alkoxy include but are not limited to isopropoxy, sec-butoxy, tert-butoxy, isopentyloxy, isohexyloxy, and the like. Examples of cyclic alkoxy include but are not limited to cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and the like. An alkoxy group can include about 1 to about 12, about 1 to about 20, or about 1 to about 40 carbon atoms bonded to the oxygen atom, and can further include double or triple bonds, and can also include heteroatoms. For example, an allyloxy group or a methoxyethoxy group is also an alkoxy group within the meaning herein, as is a methylenedioxy group in a context where two adjacent atoms of a structure are substituted therewith. The term “alkyl” as used herein refers to straight chain and branched alkyl groups and cycloalkyl groups having from 1 to 40 carbon atoms, 1 to about 20 carbon atoms, 1 to 12 carbons or, in some embodiments, from 1 to 8 carbon atoms. Examples of straight chain alkyl groups include those with from 1 to 8 carbon atoms such as methyl, ethyl, n-propyl, n-butyl, - 14 - 53224792.3 Attorney Docket No.0466483-7483WO1 n-pentyl, n-hexyl, n-heptyl, and n-octyl groups. Examples of branched alkyl groups include, but are not limited to, isopropyl, iso-butyl, sec-butyl, t-butyl, neopentyl, isopentyl, and 2,2- dimethylpropyl groups. As used herein, the term “alkyl” encompasses n-alkyl, isoalkyl, and anteisoalkyl groups as well as other branched chain forms of alkyl. Representative substituted alkyl groups can be substituted one or more times with any of the groups listed herein, for example, amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups. The term “alkynyl” as used herein refers to straight and branched chain alkyl groups, except that at least one triple bond exists between two carbon atoms. Thus, alkynyl groups have from 2 to 40 carbon atoms, 2 to about 20 carbon atoms, or from 2 to 12 carbons or, in some embodiments, from 2 to 8 carbon atoms. Examples include, but are not limited to – C^CH, -C^C(CH3), -C^C(CH2CH3), -CH2C^CH, -CH2C^C(CH3), and -CH2C^C(CH2CH3) among others. The term “alkylene” or “alkylenyl” as used herein refers to a bivalent saturated aliphatic radical (e.g., -CH2-, -CH2CH2-, and -CH2CH2CH2-, inter alia). In certain embodiments, the term may be regarded as a moiety derived from an alkene by opening of the double bond or from an alkane by removal of two hydrogen atoms from the same (e.g., - CH2-) different (e.g., -CH2CH2-) carbon atoms. Similarly, the terms “heteroalkylenyl”, “cycloalkylenyl”, “heterocycloalkylenyl”, and the like, as used herein, refer to a divalent radical of the moiety corresponding to the base group (e.g., heteroalkyl, cycloalkyl, and / or heterocycloalkyl). A divalent radical possesses two open valencies at any position(s) of the group, wherein each radical may be on a carbon atom or heteroatom. Thus, the divalent radical may form a single bond to two distinct atoms or groups, or may form a double bond with one atom. The term “antigen” or “Ag” as used herein is defined as a molecule that provokes an adaptive immune response. This immune response may involve either antibody production, or the activation of specific immunogenically-competent cells, or both. The skilled artisan will understand that any macromolecule, including virtually all proteins or peptides, can serve as an antigen. Furthermore, antigens can be derived from recombinant or genomic DNA or RNA. A skilled artisan will understand that any DNA or RNA, which comprises a nucleotide sequences or a partial nucleotide sequence encoding a protein that elicits an adaptive immune response therefore encodes an “antigen” as that term is used herein. Furthermore, one skilled in the art will understand that an antigen need not be encoded solely by a full length nucleotide sequence of a gene. It is readily apparent that the present invention includes, but is - 15 - 53224792.3 Attorney Docket No.0466483-7483WO1 not limited to, the use of partial nucleotide sequences of more than one gene and that these nucleotide sequences are arranged in various combinations to elicit the desired immune response. Moreover, a skilled artisan will understand that an antigen need not be encoded by a “gene” at all. It is readily apparent that an antigen can be generated synthesized or can be derived from a biological sample. Such a biological sample can include, but is not limited to a tissue sample, a tumor sample, a cell or a biological fluid. The term “amine” as used herein refers to primary, secondary, and tertiary amines having, e.g., the formula N(group)3wherein each group can independently be H or non-H, such as alkyl, aryl, and the like. Amines include but are not limited to R-NH2, for example, alkylamines, arylamines, alkylarylamines; R2NH wherein each R is independently selected, such as dialkylamines, diarylamines, aralkylamines, heterocyclylamines and the like; and R3N wherein each R is independently selected, such as trialkylamines, dialkylarylamines, alkyldiarylamines, triarylamines, and the like. The term “amine” also includes ammonium ions as used herein. The term “amino group” as used herein refers to a substituent of the form -NH2, - NHR, -NR2, -NR3+, wherein each R is independently selected, and protonated forms of each, except for -NR3+, which cannot be protonated. Accordingly, any compound substituted with an amino group can be viewed as an amine. An “amino group” within the meaning herein can be a primary, secondary, tertiary, or quaternary amino group. An “alkylamino” group includes a monoalkylamino, dialkylamino, and trialkylamino group. The term “anionic lipid” refers to any lipid that is negatively charged at physiological pH. These lipids include phosphatidylglycerol, cardiolipin, diacylphosphatidylserine, diacylphosphatidic acid, N-dodecanoylphosphatidylethanolamines, N- succinylphosphatidylethanolamines, N-glutarylphosphatidylethanolamines, lysylphosphatidylglycerols, palmitoyloleyolphosphatidylglycerol (POPG), and other anionic modifying groups joined to neutral lipids. The term “aryl” as used herein refers to cyclic aromatic hydrocarbon groups that do not contain heteroatoms in the ring. Thus aryl groups include, but are not limited to, phenyl, azulenyl, heptalenyl, biphenyl, indacenyl, fluorenyl, phenanthrenyl, triphenylenyl, pyrenyl, naphthacenyl, chrysenyl, biphenylenyl, anthracenyl, and naphthyl groups. In some embodiments, aryl groups contain about 6 to about 14 carbons in the ring portions of the groups. Aryl groups can be unsubstituted or substituted, as defined herein. Representative substituted aryl groups can be mono-substituted or substituted more than once, such as, but not limited to, a phenyl group substituted at any one or more of 2-, 3-, 4-, 5-, or 6-positions of - 16 - 53224792.3 Attorney Docket No.0466483-7483WO1 the phenyl ring, or a naphthyl group substituted at any one or more of 2- to 8-positions thereof. The term “cationic lipid” refers to any of a number of lipid species that carry a net positive charge at a selected pH, such as physiological pH (e.g., pH of about 7.0). It has been found that cationic lipids comprising alkyl chains with multiple sites of unsaturation, e.g., at least two or three sites of unsaturation, are particularly useful for forming lipid particles with increased membrane fluidity. A number of cationic lipids and related analogs, which are also useful in the present disclosure, have been described in U.S. Patent Publication Nos. 20060083780 and 20060240554; U.S. Patent Nos.5,208,036; 5,264,618; 5,279,833; 5,283,185; 5,753,613; and 5,785,992; and PCT Publication No. WO 96 / 10390, the disclosures of which are herein incorporated by reference in their entirety for all purposes. Non-limiting examples of cationic lipids are described in detail herein. In some cases, the cat-ionic lipids comprise a protonatable tertiary amine (e.g., pH titratable) head group, C18 alkyl chains, ether linkages between the head group and alkyl chains, and 0 to 3 double bonds. Such lipids include, e.g., DSDMA, DLinDMA, DLenDMA, and DODMA. The term “cycloalkyl” as used herein refers to cyclic alkyl groups such as, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl groups. In some embodiments, the cycloalkyl group can have 3 to about 8-12 ring members, whereas in other embodiments the number of ring carbon atoms range from 3 to 4, 5, 6, or 7. Cycloalkyl groups further include polycyclic cycloalkyl groups such as, but not limited to, norbornyl, adamantyl, bornyl, camphenyl, isocamphenyl, and carenyl groups, and fused rings such as, but not limited to, decalinyl, and the like. Cycloalkyl groups also include rings that are substituted with straight or branched chain alkyl groups as defined herein. Representative substituted cycloalkyl groups can be mono-substituted or substituted more than once, such as, but not limited to, 2,2-, 2,3-, 2,4- 2,5- or 2,6-disubstituted cyclohexyl groups or mono-, di- or tri-substituted norbornyl or cycloheptyl groups, which can be substituted with, for example, amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups. The term “cycloalkenyl” alone or in combination denotes a cyclic alkenyl group. A “disease” is a state of health of an animal wherein the animal cannot maintain homeostasis, and wherein if the disease is not ameliorated then the animal’s health continues to deteriorate. In contrast, a “disorder” in an animal is a state of health in which the animal is able to maintain homeostasis, but in which the animal’s state of health is less favorable than it would be in the absence of the disorder. Left untreated, a disorder does not necessarily cause a - 17 - 53224792.3 Attorney Docket No.0466483-7483WO1 further decrease in the animal’s state of health. A disease or disorder is “alleviated” if the severity of a symptom of the disease or disorder, the frequency with which such a symptom is experienced by a patient, or both, is reduced. As used herein, the terms “effective amount,” “pharmaceutically effective amount” and “therapeutically effective amount” refer to a nontoxic but sufficient amount of an agent to provide the desired biological result. That result may be reduction and / or alleviation of the signs, symptoms, or causes of a disease, or any other desired alteration of a biological system. An appropriate therapeutic amount in any individual case may be determined by one of ordinary skill in the art using routine experimentation. In particular, in the case of a mRNA, and “effective amount” or “therapeutically effective amount” of a therapeutic nucleic acid as relating to a mRNA is an amount sufficient to produce the desired effect, e.g., mRNA-directed expression of an amount of a protein that causes a desirable biological effect in the organism within which the protein is expressed. For example, in some embodiments, the expressed protein is an active form of a protein that is normally expressed in a cell type within the body, and the therapeutically effective amount of the mRNA is an amount that produces an amount of the encoded protein that is at least 50% (e.g., at least 60%, or at least 70%, or at least 80%, or at least 90%) of the amount of the protein that is normally expressed in the cell type of a healthy individual. For example, in some embodiments, the expressed protein is a protein that is normally expressed in a cell type within the body, and the therapeutically effective amount of the mRNA is an amount that produces a similar level of expression as observed in a healthy individual in an individual with aberrant expression of the protein (i.e., protein deficient individual). Suitable assays for measuring the expression of an mRNA or protein include, but are not limited to dot blots, Northern blots, in situ hybridization, ELISA, immunoprecipitation, enzyme function, as well as phenotypic assays known to those of skill in the art. The term “encode” as used herein refers to the product specified (e.g., protein and RNA) by a given sequence of nucleotides in a nucleic acid (i.e., DNA and / or RNA), upon transcription or translation of the DNA or RNA, respectively. In certain embodiments, the term “encode” refers to the RNA sequence specified by transcription of a DNA sequence. In certain embodiments, the term “encode” refers to the amino acid sequence (e.g., polypeptide or protein) specified by translation of mRNA. In certain embodiments, the term “encode” refers to the amino acid sequence specified by transcription of DNA to mRNA and subsequent translation of the mRNA encoded by the DNA sequence. In certain embodiments, - 18 - 53224792.3 Attorney Docket No.0466483-7483WO1 the encoded product may comprise a direct transcription or translation product. In certain embodiments, the encoded product may comprise post-translational modifications understood or reasonably expected by one skilled in the art. The term “fully encapsulated” indicates that the active agent or therapeutic agent in the lipid particle is not significantly degraded after exposure to serum or a nuclease or protease assay that would significantly degrade free DNA, RNA, or protein. In a fully encapsulated system, preferably less than about 25% of the active agent or therapeutic agent in the particle is degraded in a treatment that would normally degrade 100% of free active agent or therapeutic agent, more preferably less than about 10%, and most preferably less than about 5% of the active agent or therapeutic agent in the particle is degraded. In the context of nucleic acid therapeutic agents, full encapsulation may be determined by an OLIGREEN® assay. OLIGREEN® is an ultra-sensitive fluorescent nucleic acid stain for quantitating oligonucleotides and single-stranded DNA in solution (available from Invitrogen Corporation; Carlsbad, Calif.). RiboGreen is an ultra-sensitive fluorescent nucleic acid stain for quantitating RNA in solution. “Fully encapsulated” also indicates that the lipid particles are serum stable, that is, that they do not rapidly decompose into their component parts upon in vivo administration. The terms “halo,” “halogen,” or “halide” group, as used herein, by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom. The term “haloalkyl” group, as used herein, includes mono-halo alkyl groups, poly- halo alkyl groups wherein all halo atoms can be the same or different, and per-halo alkyl groups, wherein all hydrogen atoms are replaced by halogen atoms, such as fluoro. Examples of haloalkyl include trifluoromethyl, 1,1-dichloroethyl, 1,2-dichloroethyl, 1,3-dibromo-3,3- difluoropropyl, perfluorobutyl, and the like. The term “helper lipid” as used herein refers to a lipid capable of increasing the effectiveness of delivery of lipid-based particles such as cationic lipid-based particles to a target, preferably into a cell. The helper lipid can be neutral, positively charged, or negatively charged. In certain embodiments, the helper lipid is neutral or negatively charged. Non- limiting examples of helper lipids include 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphoethanolamine (DOPE), 1-palmitoyl- 2-oleoyl-sn-glycero-3phosphocholin (POPC) and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC). The term “heteroalkyl” as used herein by itself or in combination with another term, - 19 - 53224792.3 Attorney Docket No.0466483-7483WO1 means, unless otherwise stated, a non-cyclic stable straight or branched chain, or combinations thereof, including at least one carbon atom and at least one heteroatom selected from the group consisting of O, N, P, Si, and S, and wherein the nitrogen and sulfur atoms may optionally be oxidized, and the nitrogen heteroatom may optionally be quaternized. The heteroatom(s) (e.g., O, N, P, and S) may be placed at any interior position of the heteroalkyl group or at either terminal position at which the group is attached to the remainder of the molecule. The term “heteroaryl” as used herein refers to aromatic ring compounds containing 5 or more ring members, of which, one or more is a heteroatom such as, but not limited to, N, O, and S; for instance, heteroaryl rings can have 5 to about 8-12 ring members. A heteroaryl group is a variety of a heterocyclyl group that possesses an aromatic electronic structure. A heteroaryl group designated as a C2-heteroaryl can be a 5-ring with two carbon atoms and three heteroatoms, a 6-ring with two carbon atoms and four heteroatoms and so forth. Likewise a C4-heteroaryl can be a 5-ring with one heteroatom, a 6-ring with two heteroatoms, and so forth. The number of carbon atoms plus the number of heteroatoms sums up to equal the total number of ring atoms. Heteroaryl groups include, but are not limited to, groups such as pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridinyl, thiophenyl, benzothiophenyl, benzofuranyl, indolyl, azaindolyl, indazolyl, benzimidazolyl, azabenzimidazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, imidazopyridinyl, isoxazolopyridinyl, thianaphthalenyl, purinyl, xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, quinoxalinyl, and quinazolinyl groups. Heteroaryl groups can be unsubstituted, or can be substituted with groups as is discussed herein. Representative substituted heteroaryl groups can be substituted one or more times with groups such as those listed herein. Additional examples of aryl and heteroaryl groups include but are not limited to phenyl, biphenyl, indenyl, naphthyl (1-naphthyl, 2-naphthyl), N-hydroxytetrazolyl, N- hydroxytriazolyl, N-hydroxyimidazolyl, anthracenyl (1-anthracenyl, 2-anthracenyl, 3- anthracenyl), thiophenyl (2-thienyl, 3-thienyl), furyl (2-furyl, 3-furyl) , indolyl, oxadiazolyl, isoxazolyl, quinazolinyl, fluorenyl, xanthenyl, isoindanyl, benzhydryl, acridinyl, thiazolyl, pyrrolyl (2-pyrrolyl), pyrazolyl (3-pyrazolyl), imidazolyl (1-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl), triazolyl (1,2,3-triazol-1-yl, 1,2,3-triazol-2-yl 1,2,3-triazol-4-yl, 1,2,4-triazol-3-yl), oxazolyl (2-oxazolyl, 4-oxazolyl, 5-oxazolyl), thiazolyl (2-thiazolyl, 4- thiazolyl, 5-thiazolyl), pyridyl (2-pyridyl, 3-pyridyl, 4-pyridyl), pyrimidinyl (2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, 6-pyrimidinyl), pyrazinyl, pyridazinyl (3- pyridazinyl, 4- - 20 - 53224792.3 Attorney Docket No.0466483-7483WO1 pyridazinyl, 5-pyridazinyl), quinolyl (2-quinolyl, 3-quinolyl, 4-quinolyl, 5-quinolyl, 6- quinolyl, 7-quinolyl, 8-quinolyl), isoquinolyl (1-isoquinolyl, 3-isoquinolyl, 4-isoquinolyl, 5- isoquinolyl, 6-isoquinolyl, 7-isoquinolyl, 8-isoquinolyl), benzo[b]furanyl (2-benzo[b]furanyl, 3-benzo[b]furanyl, 4-benzo[b]furanyl, 5-benzo[b]furanyl, 6-benzo[b]furanyl, 7- benzo[b]furanyl), 2,3-dihydro-benzo[b]furanyl (2-(2,3-dihydro-benzo[b]furanyl), 3-(2,3- dihydro-benzo[b]furanyl), 4-(2,3-dihydro-benzo[b]furanyl), 5-(2,3-dihydro-benzo[b]furanyl), 6-(2,3-dihydro-benzo[b]furanyl), 7-(2,3-dihydro-benzo[b]furanyl), benzo[b]thiophenyl (2- benzo[b]thiophenyl, 3-benzo[b]thiophenyl, 4-benzo[b]thiophenyl, 5-benzo[b]thiophenyl, 6- benzo[b]thiophenyl, 7-benzo[b]thiophenyl), 2,3-dihydro-benzo[b]thiophenyl, (2-(2,3- dihydro-benzo[b]thiophenyl), 3-(2,3-dihydro-benzo[b]thiophenyl), 4-(2,3-dihydro- benzo[b]thiophenyl), 5-(2,3-dihydro-benzo[b]thiophenyl), 6-(2,3-dihydro- benzo[b]thiophenyl), 7-(2,3-dihydro-benzo[b]thiophenyl), indolyl (1-indolyl, 2-indolyl, 3-indolyl, 4-indolyl, 5-indolyl, 6-indolyl, 7-indolyl), indazole (1-indazolyl, 3-indazolyl, 4-indazolyl, 5-indazolyl, 6-indazolyl, 7-indazolyl), benzimidazolyl (1-benzimidazolyl, 2-benzimidazolyl, 4-benzimidazolyl, 5-benzimidazolyl, 6-benzimidazolyl, 7-benzimidazolyl, 8-benzimidazolyl), benzoxazolyl (1-benzoxazolyl, 2-benzoxazolyl), benzothiazolyl (1- benzothiazolyl, 2-benzothiazolyl, 4-benzothiazolyl, 5-benzothiazolyl, 6-benzothiazolyl, 7-benzothiazolyl), carbazolyl (1-carbazolyl, 2-carbazolyl, 3-carbazolyl, 4-carbazolyl), 5H-dibenz[b,f]azepine (5H-dibenz[b,f]azepin-1-yl, 5H-dibenz[b,f]azepine-2-yl, 5H-dibenz[b,f]azepine-3-yl, 5H-dibenz[b,f]azepine-4-yl, 5H-dibenz[b,f]azepine-5-yl), 10,11-dihydro-5H-dibenz[b,f]azepine (10,11-dihydro-5H-dibenz[b,f]azepine-1-yl, 10,11-dihydro-5H-dibenz[b,f]azepine-2-yl, 10,11-dihydro-5H-dibenz[b,f]azepine-3-yl, 10,11-dihydro-5H-dibenz[b,f]azepine-4-yl, 10,11-dihydro-5H-dibenz[b,f]azepine-5-yl), and the like. The term “heterocycloalkyl” as used herein refers to an aliphatic, partially unsaturated or fully saturated, 3- to 14-membered ring system, including single rings of 3 to 8 atoms and bi- and tricyclic ring systems where at least one of the carbon atoms of the ring is replaced with a heteroatom such as, but not limited to, nitrogen, oxygen, sulfur, or phosphorus. A heterocycloalkyl can include one to four heteroatoms independently selected from oxygen, nitrogen, and sulfur, wherein a nitrogen and sulfur heteroatom optionally can be oxidized and a nitrogen heteroatom optionally can be substituted. Representative heterocycloalkyl groups include, but are not limited, to the following exemplary groups: pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, and tetrahydrofuryl. - 21 - 53224792.3 Attorney Docket No.0466483-7483WO1 The term “heterocyclyl” as used herein refers to aromatic and non-aromatic ring compounds containing three or more ring members, of which one or more is a heteroatom such as, but not limited to, N, O, and S. Thus, a heterocyclyl can be a cycloheteroalkyl, or a heteroaryl, or if polycyclic, any combination thereof. In some embodiments, heterocyclyl groups include 3 to about 20 ring members, whereas other such groups have 3 to about 15 ring members. A heterocyclyl group designated as a C2-heterocyclyl can be a 5-ring with two carbon atoms and three heteroatoms, a 6-ring with two carbon atoms and four heteroatoms and so forth. Likewise a C4-heterocyclyl can be a 5-ring with one heteroatom, a 6-ring with two heteroatoms, and so forth. The number of carbon atoms plus the number of heteroatoms equals the total number of ring atoms. A heterocyclyl ring can also include one or more double bonds. A heteroaryl ring is an embodiment of a heterocyclyl group. The phrase “heterocyclyl group” includes fused ring species including those that include fused aromatic and non-aromatic groups. For example, a dioxolanyl ring and a benzdioxolanyl ring system (methylenedioxyphenyl ring system) are both heterocyclyl groups within the meaning herein. The phrase also includes polycyclic ring systems containing a heteroatom such as, but not limited to, quinuclidyl. Heterocyclyl groups can be unsubstituted, or can be substituted as discussed herein. Heterocyclyl groups include, but are not limited to, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridinyl, thiophenyl, benzothiophenyl, benzofuranyl, dihydrobenzofuranyl, indolyl, dihydroindolyl, azaindolyl, indazolyl, benzimidazolyl, azabenzimidazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, imidazopyridinyl, isoxazolopyridinyl, thianaphthalenyl, purinyl, xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, quinoxalinyl, and quinazolinyl groups. Representative substituted heterocyclyl groups can be mono-substituted or substituted more than once, such as, but not limited to, piperidinyl or quinolinyl groups, which are 2-, 3-, 4-, 5-, or 6- substituted, or disubstituted with groups such as those listed herein. The term “hydrocarbon” or “hydrocarbyl” as used herein refers to a molecule or functional group that includes carbon and hydrogen atoms. The term can also refer to a molecule or functional group that normally includes both carbon and hydrogen atoms but wherein all the hydrogen atoms are substituted with other functional groups. As used herein, the term “hydrocarbyl” refers to a functional group derived from a straight chain, branched, or cyclic hydrocarbon, and can be alkyl, alkenyl, alkynyl, aryl, cycloalkyl, acyl, or any combination thereof. Hydrocarbyl groups can be shown as (Ca- Cb)hydrocarbyl, wherein a and b are integers and mean having any of a to b number of carbon - 22 - 53224792.3 Attorney Docket No.0466483-7483WO1 atoms. For example, (C1-C4)hydrocarbyl means the hydrocarbyl group can be methyl (C1), ethyl (C2), propyl (C3), or butyl (C4), and (C0-Cb)hydrocarbyl means in certain embodiments there is no hydrocarbyl group. The term “independently selected from” as used herein refers to referenced groups being the same, different, or a mixture thereof, unless the context clearly indicates otherwise. Thus, under this definition, the phrase “X1, X2, and X3are independently selected from noble gases” would include the scenario where, for example, X1, X2, and X3are all the same, where X1, X2, and X3are all different, where X1and X2are the same but X3is different, and other analogous permutations. The term “ionizable lipid” as used herein refers to a lipid (e.g., a cationic lipid) or lipidoid having at least one protonatable or deprotonatable group, such that the lipid is positively charged at a pH at or below physiological pH (e.g., pH 7.4), and neutral at a second pH, preferably at or above physiological pH. It will be understood by one of ordinary skill in the art that the addition or removal of protons as a function of pH is an equilibrium process, and that the reference to a charged or neutral lipid refers to the nature of the predominant species and does not require that all of the lipid be present in the charged or neutral form. Generally, ionizable lipids have a pKa of the protonatable group in the range of about 4 to about 7. The term “local delivery,” as used herein, refers to delivery of an active agent or therapeutic agent such as a messenger RNA directly to a target site within an organism. For example, an agent can be locally delivered by direct injection into a disease site such as a tumor or other target site such as a site of inflammation or a target organ such as the liver, heart, pancreas, kidney, and the like. The term “lipid” refers to a group of organic compounds that include, but are not limited to, esters of fatty acids and are characterized by being insoluble in water, but soluble in many organic solvents. They are usually divided into at least three classes: (1) “simple lipids,” which include fats and oils as well as waxes; (2) “compound lipids,” which include phospholipids and glycolipids; and (3) “derived lipids” such as steroids. As used herein, “lipid encapsulated” can refer to a lipid particle that provides an active agent or therapeutic agent, such as a nucleic acid (e.g., a protein cargo), with full encapsulation, partial encapsulation, or both. In a preferred embodiment, the nucleic acid is fully encapsulated in the lipid particle (e.g., to form an SPLP, pSPLP, SNALP, or other nucleic acid-lipid particle). The term “lipid nanoparticle” refers to a particle having at least one dimension on the - 23 - 53224792.3 Attorney Docket No.0466483-7483WO1 order of nanometers (e.g., 1-1,000 nm) which includes one or more lipids and / or additional agents. The term “lipid particle” is used herein to refer to a lipid formulation that can be used to deliver an active agent or therapeutic agent, such as a nucleic acid (e.g., mRNA), to a target site of interest. In the lipid particle of the disclosure, which is typically formed from a cationic lipid, a non-cationic lipid, and a conjugated lipid that prevents aggregation of the particle, the active agent or therapeutic agent may be encapsulated in the lipid, thereby protecting the agent from enzymatic degradation. The term “monovalent” as used herein refers to a substituent connecting via a single bond to a substituted molecule. When a substituent is monovalent, such as, for example, F or Cl, it is bonded to the atom it is substituting by a single bond. The term “mRNA” or “messenger RNA” as used herein refers to a ribonucleic acid sequences which encodes a peptide or protein. In certain embodiments, the mRNA may comprise a “transcript” that is produced by using a DNA template and encodes a peptide or protein. Typically, mRNA comprises 5’-UTR, protein coding region and 3’-UTR. mRNA can be produced by in vitro transcription from a DNA template. Methods of in vitro transcription are known to those of skill in the art. For example, various in vitro transfer kits are commercially available. According to the present invention, mRNA can be modified by further stabilizing modifications and cap formation in addition to the modifications according to the invention. The term “neutral lipid” or “helper lipid” refers to any of a number of lipid species that exist either in an uncharged or neutral zwitterionic form at a selected pH. At physiological pH, such lipids include, for example, diacylphosphatidylcholine, diacylphosphatidylethanolamine, ceramide, sphingomyelin, cephalin, cholesterol, cerebrosides, and diacylglycerols. The term “non-cationic lipid” refers to any amphipathic lipid as well as any other neutral lipid or anionic lipid. The term “nucleic acid” as used herein refers to a polymer containing at least two deoxyribonucleotides or ribonucleotides in either single- or double-stranded form and includes DNA and RNA. DNA may be in the form of, e.g., antisense molecules, plasmid DNA, pre-condensed DNA, a PCR product, vectors (Pl, PAC, BAC, YAC, artificial chromosomes), expression cassettes, chimeric sequences, chromosomal DNA, or derivatives and combinations of these groups. RNA may be in the form of siRNA, asymmetrical interfering RNA (aiRNA), microRNA (miRNA), mRNA, tRNA, rRNA, tRNA, viral RNA - 24 - 53224792.3 Attorney Docket No.0466483-7483WO1 (vRNA), and combinations thereof. Nucleic acids include nucleic acids containing known nucleotide analogs or modified backbone residues or linkages, which are synthetic, naturally occurring, and non-naturally occurring, and which have similar binding properties as the reference nucleic acid. Examples of such analogs include, without limitation, phosphorothioates, phosphoramidates, methyl phosphonates, chiral-methyl phosphonates, 2’- O-methyl ribonucleotides, and peptide-nucleic acids (PNAs). Unless specifically limited, the term encompasses nucleic acids containing known analogues of natural nucleotides that have similar binding properties as the reference nucleic acid. Unless otherwise indicated, a particular nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (e.g., degenerate codon substitutions), alleles, orthologs, SNPs, and complementary sequences as well as the sequence explicitly indicated. Specifically, degenerate codon substitutions may be achieved by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed-base and / or deoxyinosine residues (Batzer et al., Nucleic Acid Res., 19:5081 (1991); Ohtsuka et al., J. Biol. Chem., 260:2605-2608 (1985); Rossolini et al., Mal. Cell. Probes, 8:91-98 (1994)). “Nucleotides” contain a sugar deoxyribose (DNA) or ribose (RNA), a base, and a phosphate group. Nucleotides are linked together through the phosphate groups. “Bases” include purines and pyrimidines, which further include natural compounds adenine, thymine, guanine, cytosine, uracil, inosine, and natural analogs, and synthetic derivatives of purines and pyrimidines, which include, but are not limited to, modifications which place new reactive groups such as, but not limited to, amines, alcohols, thiols, carboxylates, and alkyl halides. Unless otherwise indicated, a particular nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (e.g., degenerate codon substitutions), alleles, orthologs, SNPs, and complementary sequences as well as the sequence explicitly indicated. Specifically, degenerate codon substitutions may be achieved by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed-base and / or deoxyinosine residues (Batzer et al., Nucleic Acid Res., 19:5081 (1991); Ohtsuka et al., J. Biol. Chem., 260:2605-2608 (1985); Rossolini et al., Mol. Cell. Probes, 8:91-98 (1994)). The terms “patient,” “subject,” or “individual” are used interchangeably herein, and refer to any animal, or cells thereof whether in vitro or in situ, amenable to the methods described herein. In a non-limiting embodiment, the patient, subject or individual is a human. As used herein, the term “pharmaceutically acceptable” refers to a material, such as a - 25 - 53224792.3 Attorney Docket No.0466483-7483WO1 carrier or diluent, which does not abrogate the biological activity or properties of the compound, and is relatively non-toxic, i.e., the material may be administered to an individual without causing undesirable biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained. As used herein, the language “pharmaceutically acceptable salt” refers to a salt of the administered compounds prepared from pharmaceutically acceptable non-toxic acids or bases, including inorganic acids or bases, organic acids or bases, solvates, hydrates, or clathrates thereof. Suitable pharmaceutically acceptable acid addition salts may be prepared from an inorganic acid or from an organic acid. Examples of inorganic acids include hydrochloric, hydrobromic, hydriodic, nitric, carbonic, sulfuric (including sulfate and hydrogen sulfate), and phosphoric acids (including hydrogen phosphate and dihydrogen phosphate). Appropriate organic acids may be selected from aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic and sulfonic classes of organic acids, examples of which include formic, acetic, propionic, succinic, glycolic, gluconic, lactic, malic, tartaric, citric, ascorbic, glucuronic, maleic, malonic, saccharin, fumaric, pyruvic, aspartic, glutamic, benzoic, anthranilic, 4-hydroxybenzoic, phenylacetic, mandelic, embonic (pamoic), methanesulfonic, ethanesulfonic, benzenesulfonic, pantothenic, trifluoromethanesulfonic, 2- hydroxyethanesulfonic, p-toluenesulfonic, sulfanilic, cyclohexylaminosulfonic, stearic, alginic, β-hydroxybutyric, salicylic, galactaric and galacturonic acid. Suitable pharmaceutically acceptable base addition salts of compounds described herein include, for example, ammonium salts, metallic salts including alkali metal, alkaline earth metal and transition metal salts such as, for example, calcium, magnesium, potassium, sodium and zinc salts. Pharmaceutically acceptable base addition salts also include organic salts made from basic amines such as, for example, N,N’-dibenzylethylene-diamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine) and procaine. All of these salts may be prepared from the corresponding compound by reacting, for example, the appropriate acid or base with the compound. As used herein, the term “pharmaceutically acceptable carrier” or “pharmaceutically acceptable excipient” means a pharmaceutically acceptable material, composition or carrier, such as a liquid or solid filler, stabilizer, dispersing agent, suspending agent, diluent, excipient, thickening agent, solvent or encapsulating material, involved in carrying or transporting a compound described herein within or to the patient such that it may perform its intended function. Typically, such compounds are carried or transported from one organ, or - 26 - 53224792.3 Attorney Docket No.0466483-7483WO1 portion of the body, to another organ, or portion of the body. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation, including the compound(s) described herein, and not injurious to the patient. Some examples of materials that may serve as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; surface active agents; alginic acid; pyrogen-free water; isotonic saline; Ringer’s solution; ethyl alcohol; phosphate buffer solutions; and other non-toxic compatible substances employed in pharmaceutical formulations. As used herein, “pharmaceutically acceptable carrier” also includes any and all coatings, antibacterial and antifungal agents, and absorption delaying agents, and the like that are compatible with the activity of the compound(s) described herein, and are physiologically acceptable to the patient. Supplementary active compounds may also be incorporated into the compositions. The “pharmaceutically acceptable carrier” may further include a pharmaceutically acceptable salt of the compound(s) described herein. Other additional ingredients that may be included in the pharmaceutical compositions used with the methods or compounds described herein are known in the art and described, for example in Remington’s Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton, PA), which is incorporated herein by reference. The terms “peptide,” “polypeptide,” and “protein” are used interchangeably herein, and refer to a compound comprised of amino acid residues covalently linked by peptide bonds. A protein or peptide must contain at least two amino acids, and no limitation is placed on the maximum number of amino acids that can comprise a protein’s or peptide’s sequence. Polypeptides include any peptide or protein comprising two or more amino acids joined to each other by peptide bonds. As used herein, the term refers to both short chains, which also commonly are referred to in the art as peptides, oligopeptides and oligomers, for example, and to longer chains, which generally are referred to in the art as proteins, of which there are many types. “Polypeptides” include, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, variants of polypeptides, modified polypeptides, derivatives, analogs, fusion proteins, among others. The - 27 - 53224792.3 Attorney Docket No.0466483-7483WO1 polypeptides include natural peptides, recombinant peptides, synthetic peptides, or a combination thereof. The terms “polymer-conjugated lipid” and “conjugated lipid” are used interchangeably herein to refer to a lipid which is conjugated to one or more polymeric groups, which inhibits aggregation of lipid particles. Such lipid conjugates include, but are not limited to, 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 (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. By the term “specifically binds,” as used herein with respect to an antibody, is meant an antibody which recognizes a specific antigen, but does not substantially recognize or bind other molecules in a sample. For example, an antibody that specifically binds to an antigen from one species may also bind to that antigen from one or more other species. But, such cross-species reactivity does not itself alter the classification of an antibody as specific. In another example, an antibody that specifically binds to an antigen may also bind to different allelic forms of the antigen. However, such cross reactivity does not itself alter the classification of an antibody as specific. In some instances, the terms “specific binding” or “specifically binding,” can be used in reference to the interaction of an antibody, a protein, or a peptide with a second chemical species, to mean that the interaction is dependent upon the presence of a particular structure (e.g., an antigenic determinant or epitope) on the chemical species; for example, an antibody recognizes and binds to a specific protein structure rather than to proteins generally. If an antibody is specific for epitope “A”, the presence of a molecule containing epitope A (or free, unlabeled A), in a reaction containing labeled “A” and the antibody, will reduce the amount of labeled A bound to the antibody. The term “substantially” as used herein refers to a majority of, or mostly, as in at least about 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, 99.99%, or at least about 99.999% or more, or 100%. The term “substantially free of” as used herein can mean having none or having a trivial amount of, such that the amount of material present does not affect the material properties of the composition including the material, such that the - 28 - 53224792.3 Attorney Docket No.0466483-7483WO1 composition is about 0 wt% to about 5 wt% of the material, or about 0 wt% to about 1 wt%, or about 5 wt% or less, or less than, equal to, or greater than about 4.5 wt%, 4, 3.5, 3, 2.5, 2, 1.5, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.01, or about 0.001 wt% or less. The term “substantially free of” can mean having a trivial amount of, such that a composition is about 0 wt% to about 5 wt% of the material, or about 0 wt% to about 1 wt%, or about 5 wt% or less, or less than, equal to, or greater than about 4.5 wt%, 4, 3.5, 3, 2.5, 2, 1.5, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.01, or about 0.001 wt% or less, or about 0 wt%. The term “does not substantially,” when used in conjunction with a process or event indicates that the process or event does not occur or occurs in a trivial amount, such that the process or event occurs about 0% or less than about 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, or about 5.0% of the maximal amount that the process or event could occur. The term “substituted” as used herein in conjunction with a molecule or an organic group as defined herein refers to the state in which one or more hydrogen atoms contained therein are replaced by one or more non-hydrogen atoms. The term “functional group” or “substituent” as used herein refers to a group that can be or is substituted onto a molecule or onto an organic group. Examples of substituents or functional groups include, but are not limited to, a halogen (e.g., F, Cl, Br, and I); an oxygen atom in groups such as hydroxy groups, alkoxy groups, aryloxy groups, aralkyloxy groups, oxo(carbonyl) groups, carboxyl groups including carboxylic acids, carboxylates, and carboxylate esters; a sulfur atom in groups such as thiol groups, alkyl and aryl sulfide groups, sulfoxide groups, sulfone groups, sulfonyl groups, and sulfonamide groups; a nitrogen atom in groups such as amines, hydroxyamines, nitriles, nitro groups, N-oxides, hydrazides, azides, and enamines; and other heteroatoms in various other groups. Non-limiting examples of substituents that can be bonded to a substituted carbon (or other) atom include F, Cl, Br, I, OR, OC(O)N(R)2, CN, NO, NO2, ONO2, azido, CF3, OCF3, R, O (oxo), S (thiono), C(O), S(O), methylenedioxy, ethylenedioxy, N(R)2, SR, SOR, SO2R, SO2N(R)2, SO3R, C(O)R, C(O)C(O)R, C(O)CH2C(O)R, C(S)R, C(O)OR, OC(O)R, C(O)N(R)2, OC(O)N(R)2, C(S)N(R)2, (CH2)0-2N(R)C(O)R, (CH2)0-2N(R)N(R)2, N(R)N(R)C(O)R, N(R)N(R)C(O)OR, N(R)N(R)CON(R)2, N(R)SO2R, N(R)SO2N(R)2, N(R)C(O)OR, N(R)C(O)R, N(R)C(S)R, N(R)C(O)N(R)2, N(R)C(S)N(R)2, N(COR)COR, N(OR)R, C(=NH)N(R)2, C(O)N(OR)R, and C(=NOR)R, wherein R can be hydrogen or a carbon-based moiety; for example, R can be hydrogen, (C1- C100) hydrocarbyl, alkyl, acyl, cycloalkyl, aryl, aralkyl, heterocyclyl, heteroaryl, or heteroarylalkyl; or wherein two R groups bonded to a nitrogen atom or to adjacent nitrogen atoms can together with the nitrogen atom or atoms form a heterocyclyl. - 29 - 53224792.3 Attorney Docket No.0466483-7483WO1 A “therapeutic” treatment is a treatment administered to a subject who exhibits signs of pathology, for the purpose of diminishing or eliminating those signs. The term “therapeutic cargo” as used herein refers to any molecule (e.g., small molecules and macromolecules) or compound that provides a therapeutic or functional benefit to the targeted cell or tissue when delivered. The term “therapeutic protein” as used herein refers to a protein or peptide which has a positive or advantageous effect on a condition or disease state of a subject when provided to the subject in a therapeutically effective amount. In some embodiments, a therapeutic protein or peptide has curative or palliative properties and may be administered to ameliorate, relieve, alleviate, reverse, delay onset of or lessen the severity of one or more symptoms of a disease or disorder. A therapeutic protein or peptide may have prophylactic properties and may be used to delay the onset of a disease or to lessen the severity of such disease or pathological condition. The term “therapeutic protein” includes entire proteins or peptides, and can also refer to therapeutically active fragments thereof. It can also include therapeutically active variants of a protein. Exemplary therapeutic proteins include, but are not limited to, an analgesic protein, an anti-inflammatory protein, an anti-proliferative protein, an proapoptotic protein, an anti-angiogenic protein, a cytotoxic protein, a cytostatic protein, a cytokine, a chemokine, a growth factor, a wound healing protein, a pharmaceutical protein, or a pro-drug activating protein. Therapeutic proteins may include growth factors (EGF, TGF-α, TGF- β, TNF, HGF, IGF, and IL-1-8, inter alia) cytokines, paratopes, Fabs (fragments, antigen binding), and antibodies. The terms “treat,” “treating” and “treatment,” as used herein, means reducing the frequency or severity with which symptoms of a disease or condition are experienced by a subject by virtue of administering an agent or compound to the subject. Aryl-Alkyl Disulfide Ionizable Lipid Compounds In one aspect, the disclosure provides a compound of formula (I), or a salt, stereoisomer, or isotopologue thereof: (I), wherein: A is selected from the group consisting of ; - 30 - 53224792.3 Attorney Docket No.0466483-7483WO1 each occurrence of L1, if present, is independently selected from the group consisting of -(optionally substituted C1-C12alkylenyl)-, -(optionally substituted C2-C12alkenylenyl)-, - (optionally substituted C1-C12 alkynylenyl)-, -(optionally substituted C1-C12 heteroalkylenyl)- , -(optionally substituted C3-C8cycloalkylenyl)-, -(optionally substituted C2-C8heterocyloalkylenyl)-, -(optionally substituted C6-C10 arylenyl)-, -(optionally substituted C2- C8heteroarylenyl)-, -(optionally substituted C1-C12alkylenyl)-X-, -(optionally substituted C2- C12 alkenylenyl)-X-, -(optionally substituted C1-C12 alkynylenyl)-X-, -(optionally substituted C1-C12heteroalkylenyl)-X-, -(optionally substituted C3-C8cycloalkylenyl)-X-, -(optionally substituted C2-C8 heterocyloalkylenyl)-X-, -(optionally substituted C6-C10 arylenyl)-X-, and - (optionally substituted C2-C8heteroarylenyl)-X-; each occurrence of X, if present, is independently selected from the group consisting of -N(R1e)-, -N(RA)-, and -O-; each occurrence of R1a, R1b, R1c, R1d, and R1e, if present, is independently , wherein: R1cand R1dcan combine with the atoms to which they are bound to form an optionally substituted C2-C8 heterocycloalkyl, or one of R1cand R1dcan combine with one occurrence of L1to form an optionally substituted C2-C8heterocycloalkyl; R2is selected from the group consisting of optionally substituted C1-C6 alkyl, optionally substituted C2-C6alkenyl, optionally substituted C3-C8cycloalkyl, optionally substituted C2-C6 heteroalkyl, optionally substituted C3-C6 heteroalkenyl, optionally substituted C2-C8heterocycloalkyl, and N(RA)(RB); each occurrence of L2is independently selected from the group consisting of - (optionally substituted C1-C6alkylenyl)-, -C(=O)-, -O-, and -N(RA)-; each occurrence of R3is independently selected from the group consisting of optionally substituted C1-C24alkyl and optionally substituted C1-C24heteroalkyl; each occurrence of Ar is independently selected from the group consisting of - (optionally substituted C6-C10heteroarylenyl)- and -(optionally substituted C2-C8heteroarylenyl)-; m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; and each occurrence of RAand RBis independently selected from the group consisting of H, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally - 31 - 53224792.3 Attorney Docket No.0466483-7483WO1 substituted C3-C8 cycloalkyl, optionally substituted C2-C6 heteroalkyl, optionally substituted C3-C6heteroalkenyl, optionally substituted C2-C8heterocycloalkyl, optionally substituted C6- C10 aryl, and optionally substituted C2-C8 heteroaryl. In certain embodiments, L1is -(CH2)1-5-. In certain embodiments, L1is -(CH2)1-5N(R1e)-. In certain embodiments, L1is . (Ia): certain embodiments, the compound of formula (I) is a compound of In certain embodiments, the compound of formula (I) is a compound of formula (Ic). In certain embodiments, L2is -(CH2)1-5-. In certain embodiments, L2is -CH(OH)-. In certain embodiments, L2is -O-. In certain embodiments, each occurrence of R1a, R1b, R1c, R1d, and R1e, if present, is , wherein: each selected from the group consisting of optionally substituted C1-C6alkyl, optionally substituted C2-C6alkenyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C6 heteroalkyl, optionally substituted C3-C6 heteroalkenyl, optionally substituted C2-C8heterocycloalkyl, optionally substituted C6-C10aryl, optionally substituted C2-C8 heteroaryl, ORA, N(RA)(RB), halogen, CN, NO2, C(=O)RA, - 32 - 53224792.3 Attorney Docket No.0466483-7483WO1 C(=O)ORA, C(=O)N(RA)(RB), S(=O)2RA, S(=O)2ORA, and S(=O)2N(RA)(RB), wherein two vicinal R4moieties can combine with the carbon atoms to which they are bound to form an optionally substituted C3-C8 cycloalkyl, C2- C8heterocycloalkyl, C6-C10aryl, or C2-C8heteroaryl; and o is 0, 1, 2, 3, or 4. In certain embodiments, -(L2)n- is . In certain embodiments, -(L2)n- is . In certain embodiments, - n- . In certain In certain embodiments, R1ais embodiments, R1a. In certain embodiments, R1ais In certain embodiments, R1a. In certain embodiments, R1ais In certain In certain embodiments, R1bis - 33 - 53224792.3 Attorney Docket No.0466483-7483WO1 embodiments, R1b. In certain embodiments, R1bis In certain embodiments, R1b. In certain embodiments, R1bis In certain embodiments, R1c. In certain embodiments, R1cis embodiments, R1c. In certain embodiments, R1cis In certain embodiments, R1c. In certain embodiments, R1cis In certain In certain embodiments, R1dis - 34 - 53224792.3 Attorney Docket No.0466483-7483WO1 embodiments, R1d. In certain embodiments, R1dis In certain embodiments, R1d. In certain embodiments, R1dis In certain embodiments, R1e. In certain embodiments, R1eis embodiments, R1e. In certain embodiments, R1eis In certain embodiments, R1e. In certain embodiments, R1eis . - 35 - 53224792.3 Attorney Docket No.0466483-7483WO1 In certain embodiments, R3is optionally substituted C4-C12 alkyl. In certain embodiments, R3is butyl. In certain embodiments, R3is pentyl. In certain embodiments, R3is hexyl. In certain embodiments, R3is heptyl. In certain embodiments, R3is octyl. In certain embodiments, R3is nonyl. In certain embodiments, R3is decyl. In certain embodiments, R3is undecyl. In certain embodiments, R3is dodecyl. In certain embodiments, R3is n-butyl. In certain embodiments, R3is n-pentyl. In certain embodiments, R3is n-hexyl. In certain embodiments, R3is n-heptyl. In certain embodiments, R3is n-octyl. In certain embodiments, R3is n-nonyl. In certain embodiments, R3is n-decyl. In certain embodiments, R3is n- undecyl. In certain embodiments, R3is n-dodecyl. In certain embodiments, R1a. In certain embodiments, certain embodiments, R1ais In certain embodiments, R1a. In certain embodiments, R1ais In In certain embodiments, R1a. In certain embodiments, R1ais - 36 - 53224792.3 Attorney Docket No.0466483-7483WO1 OH O S S In . In certain embodiments, R1a. In certain embodiments, R1ais . In certain embodiments, R1a. In certain embodiments, R1ais . In certain . In certain embodiments, R1ais In certain embodiments, R1a. In certain embodiments, R1ais In certain embodiments, R1a. In certain embodiments, R1ais In - 37 - 53224792.3 Attorney Docket No.0466483-7483WO1 certain embodiments, R1ais . In certain embodiments, R1ais OH O S is is is . In certain embodiments, R1bis In certain embodiments, R1b. In certain embodiments, R1bis In In certain embodiments, R1b. In certain embodiments, R1bis - 38 - 53224792.3 Attorney Docket No.0466483-7483WO1 OH O S S In certain embodiments, R1b. In certain embodiments, R1bis . In certain embodiments, R1b. In certain embodiments, R1bis . In certain embodiments, R1b. In certain embodiments, R1bis . In certain embodiments, R1b. In certain embodiments, R1bis In certain embodiments, R1b. In certain embodiments, R1bis In certain embodiments, R1b. In certain embodiments, R1bis In - 39 - 53224792.3 Attorney Docket No.0466483-7483WO1 certain embodiments, R1bis . In certain embodiments, R1bis OH O S is is . In certain embodiments, R1cis In certain embodiments, R1c. In certain embodiments, R1cis In certain embodiments, R1c. In certain embodiments, R1cis In - 40 - 53224792.3 Attorney Docket No.0466483-7483WO1 OH O S S In . In certain embodiments, R1c. In certain embodiments, R1cis . In certain embodiments, R1c. In certain embodiments, R1cis . In certain . In certain embodiments, R1cis In certain embodiments, R1c. In certain embodiments, R1cis In certain embodiments, R1c. In certain embodiments, R1cis In - 41 - 53224792.3 Attorney Docket No.0466483-7483WO1 certain embodiments, R1cis . In certain embodiments, R1cis OH O S is is is . In certain embodiments, R1dis In certain embodiments, R1d. In certain embodiments, R1dis In certain embodiments, R1d. In certain embodiments, R1dis In certain embodiments, R1d. In certain embodiments, R1dis - 42 - 53224792.3 Attorney Docket No.0466483-7483WO1 OH O S S In certain embodiments, R1d. In certain embodiments, R1dis . In certain embodiments, R1d. In certain embodiments, R1dis . In certain embodiments, R1d. In certain embodiments, R1dis . In certain embodiments, R1d. In certain embodiments, R1dis In certain embodiments, R1d. In certain embodiments, R1dis In certain embodiments, R1d. In certain embodiments, R1dis In - 43 - 53224792.3 Attorney Docket No.0466483-7483WO1 certain embodiments, R1dis . In certain embodiments, R1dis OH O S is is . In certain embodiments, R1eis In certain embodiments, R1e. In certain embodiments, R1eis In certain embodiments, R1e. In certain embodiments, R1eis In - 44 - 53224792.3 Attorney Docket No.0466483-7483WO1 OH O S S In . In certain embodiments, R1e. In certain embodiments, R1eis . In certain embodiments, R1e. In certain embodiments, R1eis . In certain . In certain embodiments, R1eis In certain embodiments, R1e. In certain embodiments, R1eis In certain embodiments, R1e. In certain embodiments, R1eis In - 45 - 53224792.3 Attorney Docket No.0466483-7483WO1 certain embodiments, R1eis . In certain embodiments, R1eis OH O S is is the group consisting of: 3,3'-((2-(4-(2-((2-(bis(3-(2-(hexyldisulfaneyl)phenoxy)-2- C6 PhE 200 hydroxypropyl)amino)ethyl)(3-(2-(hexyldisulfaneyl)phenoxy)-2- - 46 - 53224792.3 Attorney Docket No.0466483-7483WO1 200 hydroxypropyl)amino)ethyl)(3-(3-(dodecyldisulfaneyl)phenoxy)-2- hydroxypropyl)amino)ethyl)piperazin-1-yl)ethyl)azanediyl)bis(1-(3- (dodecldisulfanel)henox)roan-2-ol) 1- 1- 1- ); - 47 - 53224792.3 Attorney Docket No.0466483-7483WO1 diyl))bis(azanetriyl))tetrakis(1-(2-(octyldisulfaneyl)phenoxy)propan-2-ol); C10oPhE- 3,3',3'',3'''-((piperazine-1,4-diylbis(propane-3,1- 383 dil))bi(zntril))ttrki(1-(2-(d ldi lf n l)hnx)r n-2-l); - ); ); ); - ); ); ); - n ceran emo mens, eac occurrence o op ona y sus ue a y, op ona y substituted alkenyl, optionally substituted cycloalkyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heterocycloalkyl, optionally substituted alkylenyl, optionally substituted alkenylenyl, optionally substituted alkynylenyl, optionally substituted heteroalkylenyl, optionally substituted cycloalkylenyl, optionally substituted heterocycloalkylenyl, optionally substituted arylenyl, and optionally substituted heteroarylenyl is independently optionally substituted with at least one substituent selected from the group consisting of C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 haloalkyl, C1-C3 haloalkoxy, phenoxy, halogen, CN, NO2, OH, N(R’)(R’’), C(=O)R’, C(=O)OR’, OC(=O)OR’, C(=O)N(R’)(R’’), S(=O)2N(R’)(R’’), N(R’)C(=O)R’’, N(R’)S(=O)2R’’, C2-C8 heteroaryl, and phenyl optionally substituted with at least one halogen, wherein each occurrence of R’ and R’’ is independently selected from the group consisting of H, C1-C6 alkyl, C3-C8cycloalkyl, C1-C6haloalkyl, benzyl, and phenyl, or wherein R’ and R’’ can combine with the nitrogen atom to which they are bound to form a C2-C8 heterocycloalkyl (e.g., cyclic tertiary amine). - 48 - 53224792.3 Attorney Docket No.0466483-7483WO1 Amidine-Crosslinker Containing Ionizable Lipid Compounds In one aspect, the disclosure provides a compound of formula (II), or a salt, stereoisomer, or isotopologue thereof: , wherein: R1a, R1b, and a polyamine moiety, wherein each amine substituent of the polyamine moiety is substituted with at least one optionally substituted C1-C20 alkyl, optionally substituted C1-C20 heteroalkyl, optionally substituted C2-C20alkenyl, optionally substituted C2-C20heteroalkyenyl, optionally substituted C2-C20 alkynyl, and optionally substituted C2-C20 heteroalkynyl; R2a, R2b, and each occurrence of R2c, if present, are each independently selected from the group consisting of H and optionally substituted C1-C6 alkyl; R3a, R3b, and each occurrence of R3c, if present, are each independently selected from the group consisting of H and optionally substituted C1-C6alkyl; each occurrence of R4aand R4bis independently selected from the group consisting of H, optionally substituted C1-C6alkyl, optionally substituted C1-C6heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C6-C10aryl, and optionally substituted C2-C8heteroaryl; each occurrence of R5aand R5bis independently selected from the group consisting of H, optionally substituted C1-C6alkyl, optionally substituted C1-C6heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C6-C10aryl, and optionally substituted C2-C8heteroaryl; each occurrence of R6is ; each occurrence of R7aand R7b, if selected from the group consisting of H, optionally substituted C1-C6alkyl, optionally substituted C1-C6heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted C2-C8 heteroaryl; m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; - 49 - 53224792.3 Attorney Docket No.0466483-7483WO1 each occurrence of n is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; o is 0, 1, 2, 3, 4, or 5; and each occurrence of p is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In certain embodiments, R2ais H. In certain embodiments, R2bis H. In certain embodiments, R2cis H. In certain embodiments, R3ais H. In certain embodiments, R3bis H. In certain embodiments, R3cis H. In certain embodiments, R4ais H. In certain embodiments, R4bis H. In certain embodiments, R5ais H. In certain embodiments, R5bis H. In certain embodiments, R7ais H. In certain embodiments, R7bis H. NH H N R1bR1aN In certain embodiments, the compound of formula (II) (IIa). In certain embodiments, the compound of formula (II) (IIb). In certain embodiments, the compound of formula (II) . each occurrence of R1c, if present, are each , wherein: each if present, is independently selected from the group consisting of -(optionally substituted C1-C12alkylenyl)-, -(optionally substituted C2-C12alkenylenyl)-, - (optionally substituted C1-C12 alkynylenyl)-, -(optionally substituted C1-C12 heteroalkylenyl)- , -(optionally substituted C3-C8cycloalkylenyl)-, -(optionally substituted C2-C8heterocyloalkylenyl)-, -(optionally substituted C6-C10 arylenyl)-, -(optionally substituted C2- C8heteroarylenyl)-, -(optionally substituted C1-C12alkylenyl)-X-, -(optionally substituted C2- C12 alkenylenyl)-X-, -(optionally substituted C1-C12 alkynylenyl)-X-, -(optionally substituted C1-C12heteroalkylenyl)-X-, -(optionally substituted C3-C8cycloalkylenyl)-X-, -(optionally substituted C2-C8 heterocyloalkylenyl)-X-, -(optionally substituted C6-C10 arylenyl)-X-, - - 50 - 53224792.3 Attorney Docket No.0466483-7483WO1 (optionally substituted C2-C8 heteroarylenyl)-X-, and -X-; each occurrence of X, if present, is independently selected from the group consisting of -N(R8c)-, -N(RA)-, -O-, and -S-; each occurrence of R8a, R8b, and R8c, if present, is independently selected from the group consisting of optionally substituted C1-C20 alkyl, optionally substituted C1-C20 heteroalkyl, optionally substituted C2-C20alkenyl, optionally substituted C2-C20heteroalkyenyl, optionally substituted C2-C20 alkynyl, and optionally substituted C2-C20 heteroalkynyl, wherein one of R8aand R8bcan combine with one occurrence of L1to form an optionally substituted C2-C8heterocycloalkyl; each occurrence of RAand RBis independently selected from the group consisting of H and optionally substituted C1-C6alkyl; and each occurrence of q is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In certain embodiments, L1is -(CH2)1-5-. In certain embodiments, L1is -(CH2)1-5CH(ORA)-. In certain embodiments, L1is -(CH2)1-5O-. In certain embodiments, L1is - (CH2)1-5N(RA)-. In certain embodiments, L1is -(CH2)1-5N(R8c)-. In certain embodiments, L1is -S-. In certain embodiments, L1. In certain embodiments, L1 . In certain . In certain . In certain embodiments, -(L1)p- is . In certain embodiments, - p- . In certain embodiments, - In certain embodiments, -(L1)p- is . In certain embodiments, -(L1)p- is . In certain embodiments, -(L1)p- . In certain embodiments, -(L1)p- . In certain embodiments, -(L1)p- is - 51 - 53224792.3 Attorney Docket No.0466483-7483WO1 . . p- is p- is is R1ais - 52 - 53224792.3 Attorney Docket No.0466483-7483WO1 . In certain embodiments, R1ais . In certain embodiments, R1a. In certain embodiments, R1ais is R1ais . In certain embodiments, R1a. In certain embodiments, R1a. In certain embodiments, R1ais is In certain embodiments, R1ais - 53 - 53224792.3 Attorney Docket No.0466483-7483WO1 is is . In - 54 - 53224792.3 Attorney Docket No.0466483-7483WO1 certain embodiments, R1b. In certain embodiments, In certain embodiments, R1bis . In certain embodiments, R1b. In certain embodiments, R1b. In certain embodiments, R1bis is is is - 55 - 53224792.3 Attorney Docket No.0466483-7483WO1 R8aN R8bis is is . In - 56 - 53224792.3 Attorney Docket No.0466483-7483WO1 In certain embodiments, R1cis embodiments, R1c. In certain embodiments, R1cis is is is - 57 - 53224792.3 Attorney Docket No.0466483-7483WO1 certain embodiments, R1cis is . In certain embodiments, R8bis -(CH2)CH(OH)(optionally substituted C3-C20 alkyl). In certain embodiments, R8cis -(CH2)CH(OH)(optionally substituted C3-C20alkyl). In certain embodiments, R8ais -(CH2)CH(OH)(CH2)3CH3. In certain embodiments, R8ais -(CH2)CH(OH)(CH2)3CH3. In certain embodiments, R8ais -(CH2)CH(OH)(CH2)4CH3. In certain embodiments, R8ais -(CH2)CH(OH)(CH2)5CH3. In certain embodiments, R8ais - (CH2)CH(OH)(CH2)6CH3. In certain embodiments, R8ais -(CH2)CH(OH)(CH2)7CH3. In certain embodiments, R8ais -(CH2)CH(OH)(CH2)8CH3. In certain embodiments, R8ais - (CH2)CH(OH)(CH2)9CH3. In certain embodiments, R8ais -(CH2)CH(OH)(CH2)10CH3. In certain embodiments, R8ais -(CH2)CH(OH)(CH2)11CH3. In certain embodiments, R8ais - (CH2)CH(OH)(CH2)12CH3. In certain embodiments, R8ais -(CH2)CH(OH)(CH2)13CH3. In certain embodiments, R8ais -(CH2)CH(OH)(CH2)14CH3. In certain embodiments, R8ais - (CH2)CH(OH)(CH2)15CH3. In certain embodiments, R8ais -(CH2)CH(OH)(CH2)16CH3. In certain embodiments, R8ais -(CH2)CH(OH)(CH2)17CH3. In certain embodiments, R8ais - (CH2)CH(OH)(CH2)18CH3. In certain embodiments, R8ais -(CH2)CH(OH)(CH2)19CH3. In certain embodiments, R8ais -(CH2)CH(OH)(CH2)20CH3. In certain embodiments, R8bis - (CH2)CH(OH)(CH2)3CH3. In certain embodiments, R8bis -(CH2)CH(OH)(CH2)3CH3. In certain embodiments, R8bis -(CH2)CH(OH)(CH2)4CH3. In certain embodiments, R8bis - (CH2)CH(OH)(CH2)5CH3. In certain embodiments, R8bis -(CH2)CH(OH)(CH2)6CH3. In certain embodiments, R8bis -(CH2)CH(OH)(CH2)7CH3. In certain embodiments, R8bis - (CH2)CH(OH)(CH2)8CH3. In certain embodiments, R8bis -(CH2)CH(OH)(CH2)9CH3. In certain embodiments, R8bis -(CH2)CH(OH)(CH2)10CH3. In certain embodiments, R8bis - (CH2)CH(OH)(CH2)11CH3. In certain embodiments, R8bis -(CH2)CH(OH)(CH2)12CH3. In - 58 - 53224792.3 Attorney Docket No.0466483-7483WO1 certain embodiments, R8bis -(CH2)CH(OH)(CH2)13CH3. In certain embodiments, R8bis - (CH2)CH(OH)(CH2)14CH3. In certain embodiments, R8bis -(CH2)CH(OH)(CH2)15CH3. In certain embodiments, R8bis -(CH2)CH(OH)(CH2)16CH3. In certain embodiments, R8bis - (CH2)CH(OH)(CH2)17CH3. In certain embodiments, R8bis -(CH2)CH(OH)(CH2)18CH3. In certain embodiments, R8bis -(CH2)CH(OH)(CH2)19CH3. In certain embodiments, R8bis - (CH2)CH(OH)(CH2)20CH3. In certain embodiments, R8cis -(CH2)CH(OH)(CH2)3CH3. In certain embodiments, R8cis -(CH2)CH(OH)(CH2)3CH3. In certain embodiments, R8cis - (CH2)CH(OH)(CH2)4CH3. In certain embodiments, R8cis -(CH2)CH(OH)(CH2)5CH3. In certain embodiments, R8cis -(CH2)CH(OH)(CH2)6CH3. In certain embodiments, R8cis - (CH2)CH(OH)(CH2)7CH3. In certain embodiments, R8cis -(CH2)CH(OH)(CH2)8CH3. In certain embodiments, R8cis -(CH2)CH(OH)(CH2)9CH3. In certain embodiments, R8cis - (CH2)CH(OH)(CH2)10CH3. In certain embodiments, R8cis -(CH2)CH(OH)(CH2)11CH3. In certain embodiments, R8cis -(CH2)CH(OH)(CH2)12CH3. In certain embodiments, R8cis - (CH2)CH(OH)(CH2)13CH3. In certain embodiments, R8cis -(CH2)CH(OH)(CH2)14CH3. In certain embodiments, R8cis -(CH2)CH(OH)(CH2)15CH3. In certain embodiments, R8cis - (CH2)CH(OH)(CH2)16CH3. In certain embodiments, R8cis -(CH2)CH(OH)(CH2)17CH3. In certain embodiments, R8cis -(CH2)CH(OH)(CH2)18CH3. In certain embodiments, R8cis - (CH2)CH(OH)(CH2)19CH3. In certain embodiments, R8cis -(CH2)CH(OH)(CH2)20CH3. In certain In certain embodiments, R1a. In certain . In certain embodiments, - 59 - 53224792.3 Attorney Docket No.0466483-7483WO1 In certain . In certain embodiments, R1ais is In certain embodiments, R1ais In certain . In certain embodiments, R1ais - 60 - 53224792.3 Attorney Docket No.0466483-7483WO1 is R1ais . In certain embodiments, R1ais is is R1ais - 61 - 53224792.3 Attorney Docket No.0466483-7483WO1 is . In certain In certain embodiments, R1bis - 62 - 53224792.3 Attorney Docket No.0466483-7483WO1 In certain In certain embodiments, R1b. In certain In certain . In certain embodiments, R1bis is In certain embodiments, R1bis certain embodiments, R1bis - 63 - 53224792.3 Attorney Docket No.0466483-7483WO1 . In certain In certain embodiments, R1bis is R1bis is certain embodiments, R1bis - 64 - 53224792.3 Attorney Docket No.0466483-7483WO1 is is is is . Attorney Docket No.0466483-7483WO1 In certain In certain embodiments, R1cis . In certain In certain embodiments, In certain In certain embodiments, R1cis certain embodiments, R1cis - 66 - 53224792.3 Attorney Docket No.0466483-7483WO1 is In certain . In certain embodiments, R1cis is R1cis is Attorney Docket No.0466483-7483WO1 is is is is - 68 - 53224792.3 Attorney Docket No.0466483-7483WO1 . In certain In certain embodiments, In certain . In certain embodiments, In - 69 - 53224792.3 Attorney Docket No.0466483-7483WO1 certain . In certain embodiments, R1ais certain embodiments, R1ais is . In certain . In certain embodiments, R1ais certain embodiments, R1ais - 70 - 53224792.3 Attorney Docket No.0466483-7483WO1 certain embodiments, R1ais . embodiments, is - 71 - 53224792.3 Attorney Docket No.0466483-7483WO1 is is . In certain In certain embodiments, R1bis In - 72 - 53224792.3 Attorney Docket No.0466483-7483WO1 certain In certain embodiments, . In certain . In certain embodiments, R1bis certain embodiments, R1bis is In - 73 - 53224792.3 Attorney Docket No.0466483-7483WO1 certain . In certain embodiments, R1bis NN. C H21O C21In certain embodimen . In certain embodiments, certain embodiments, R1bis - 74 - 53224792.3 Attorney Docket No.0466483-7483WO1 is . In certain . In certain embodiments, R1cis - 75 - 53224792.3 Attorney Docket No.0466483-7483WO1 In certain In certain embodiments, . In certain In certain . In certain embodiments, R1cis is In certain embodiments, R1cis embodiments, R1cis - 76 - 53224792.3 Attorney Docket No.0466483-7483WO1 In certain . In certain embodiments, R1cis NNis is . In certain In certain is - 77 - 53224792.3 Attorney Docket No.0466483-7483WO1 is . - 78 - 53224792.3 Attorney Docket No.0466483-7483WO1 N1,N6-bis(3-(4-(3-(bis(2-hydroxydodecyl)amino)propyl)piperazin-1- yl)propyl)adipimidamide, (C12-2aN). In certain embodiments, each occurrence of optionally substituted alkyl, optionally substituted alkenyl, optionally substituted cycloalkyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heterocycloalkyl, optionally substituted alkylenyl, optionally substituted alkenylenyl, optionally substituted alkynylenyl, optionally substituted heteroalkylenyl, optionally substituted cycloalkylenyl, optionally substituted heterocycloalkylenyl, optionally substituted arylenyl, and optionally substituted heteroarylenyl is independently optionally substituted with at least one substituent selected from the group consisting of C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 haloalkyl, C1-C3 haloalkoxy, phenoxy, halogen, CN, NO2, OH, N(R’)(R’’), C(=O)R’, C(=O)OR’, OC(=O)OR’, C(=O)N(R’)(R’’), S(=O)2N(R’)(R’’), N(R’)C(=O)R’’, N(R’)S(=O)2R’’, C2-C8 heteroaryl, and phenyl optionally substituted with at least one halogen, wherein each occurrence of R’ and R’’ is independently selected from the group consisting of H, C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 haloalkyl, benzyl, and phenyl, or wherein R’ and R’’ can combine with the nitrogen atom to which they are bound to form a C2-C8 heterocycloalkyl (e.g., cyclic tertiary amine). Ionizable Lipids and / or Cationic Lipids or Lipidoids The scope of ionizable lipids contemplated for use in the present disclosure is not limited to ionizable lipidoids of formula (I) or (II). In the lipid nanoparticles of the disclosure, the cationic lipid or ionizable lipid may comprise, e.g., one or more of the following: (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl 4-(dimethylamino)butanoate (DLinMC3DMA), [(4-hydroxybutyl)azanediyl]di(hexane-6,1-diyl) bis(2-hexyldecanoate) (ALC-0315), heptadecan-9-yl 8-{(2-hydroxyethyl)[6-oxo-6- (undecyloxy)hexyl]amino}octanoate (SM-102), 1,1′-[[2-[4-[2-[[2-[bis(2- hydroxydodecyl)amino]ethyl](2-hydroxydodecyl)amino]ethyl]-1- piperazinyl]ethyl]imino]bis-2-dodecanol (C12-200), 1,2-dilinoleyloxy-N,N- dimethylaminopropane (DLinDMA), 1,2-dilinolenyloxy-N,N-dimethylaminopropane (DLenDMA), 2,2-dilinoleyl-4-(2-dimethylaminoethyl)-[1,3]-dioxolane (DLin-K-C2-DMA; “XTC2”), 2,2-dilinoleyl-4-(3- 45 dimethylaminopropyl)- 1,3]-dioxolane (D Lin-K-C3-D MA), 2,2-dilinoleyl-4-(4-dimethylaminobutyl)-[1,3]-dioxolane (DLin-K-C4-DMA), 2,2- dilinoleyl-5-dimethylaminomethyl-[1,3]-dioxane (DLin-K6-DMA), 2,2-dilinoleyl-4-N- - 79 - 53224792.3 Attorney Docket No.0466483-7483WO1 methylpepiazino-[1,3]-dioxolane (DLin-K-MPZ), 2,2-dili-noleyl-4-dimethylaminomethyl- [1,3]-dioxolane (DLin-KDMA), 1,2-dilinoleylcarbamoyloxy-3-dimethylaminopropane (D Lin-C-DAP), 1,2-dilinoleyoxy-3-(dimethylaminoacetoxypropane (DLin-DAC), 1- 2dilinoleyoxy-3-morpholinopropane (DLin-MA), 1,2-dilinoleoyl-3-dimethylaminopropane (DLinDAP), 1,2-dilinoleylthio-3-dimethylaminopropane (DLin-2-DMAP), 1,2-dilinoleyloxy- 3-trimethylaminopropane chloride salt (DLin-TMA.Cl), 1,2-dilinoleoyl-3- trimethylaminopropane chloride salt (DLin-TAP.Cl), 1,2-dilinoleyloxy-3-(N- methylpiperazino)propane (D Lin-MPZ), 3-(N,N-dilinoleylamino)-1,2-propanediol (D LinAP), 3-(N,N-dioleylamino)-1,2-propanedio (DOAP), 1,2-dilinoleyloxo-3-(2-N,N- dimethylamino)ethoxypropane (D Lin-EG-D MA), N,N-dioleyl-N,N-dimethylanrmonium chloride (DODAC), 1,2-dioleyloxy-N,N-dimethylaminopropane (DODMA), 1,2- distearyloxy-N,N-dimethylaminopropane (DSD MA), N-(1-(2,3-dioleyloxy)propyl)-N,N,N- trimethylammonium chloride (DOTMA), N,N-distearyl-N,N-dimethylammonium bromide (DDAB), N-(1-(2,3-dioleoyloxy)propyl)-N,N, N-trimethylammonium chloride (DOTAP), 3- (N-(N’,N’dimethylaminoethane)-carbamoyl)cholesterol (DC-Chol), N-(l,2- dimyristyloxyprop-3-yl)-N,N-dimethyl-N-hydroxyethyl anrmonium bromide (DMRIE), 2,3- dioleyloxy-N-[2 (spermine-carboxamidoethyl]-N,N-dimethy 1-1- propanaminiumtrifluoroacetate (DOSPA), dioctadecylamidoglycyl spermine (DOGS), 3- dimethylamino-2-(cholest-5-en-3-beta-oxybutan-4-oxy)-1-(cis,cis-9,12- octadecadienoxy)propane (CLinDMA), 2-[5’-(cholest-5-en-3-beta-oxy)-3’-oxapentoxy)-3- dimethyl-1-(cis,cis-9’,1-2’-octadecadienoxy) propane (CpLinDMA), N,N-dimethyl-3,4- dioleyloxybenzylamine (DMOBA), 1,2-N,N’dioleylcarbamyl-3-dimethylaminopropane (DOcarbDAP), 1,2-N,N’-dilinoleylcarbamyl-3-dimethylaminopropane (DLincarbDAP), or mixtures thereof. In certain embodiments, the cationic lipid is DLinDMA, DLin-K-C2-DMA (“XTC2”), or mixtures thereof. The ionizable lipids are not limited to those recited herein, and can further include ionizable lipids known to those skilled in the art, or described in PCT Application No. PCT / US2020 / 056255 and / or PCT Application No. PCT / US2020 / 056252, the disclosures of which are herein incorporated by reference in its entirety. The synthesis of cationic lipids such as DLin-K-C2-DMA (“XTC2”), DLin-K-C3- DMA, DLin-K-C4-DMA, DLin-K6-DMA, and DLin-K-MPZ, as well as additional cationic lipids, is described in U.S. Application Publication No. US 2011 / 0256175, the disclosure of which is herein incorporated by reference in its entirety for all purposes. The synthesis of cationic lipids such as DLin-K-DMA, DLin-CDAP, DLin-DAC, DLin-MA, DLinDAP, DLin-S-DMA, DLin-2-DMAP, DLin-TMA.Cl, DLin-TAP.Cl, DLin-MPZ, DLinAP, DOAP, - 80 - 53224792.3 Attorney Docket No.0466483-7483WO1 and DLin-EG-DMA, as well as additional cationic lipids, is described in PCT Application No. PCT / US08 / 88676, filed December 31, 2008, the disclosure of which is herein incorporated by reference in its entirety for all purposes. The synthesis of cationic lipids such as CLinDMA, as well as additional cationic lipids, is described in U.S. Patent Publication No. 20060240554, the disclosure of which is herein incorporated by reference in its entirety for all purposes. Non-cationic Lipid In the nucleic acid-lipid particles of the present disclosure, the non-cationic lipid may comprise, e.g., one or more anionic lipids and / or neutral lipids. In some embodiments, the non-cationic lipid comprises one of the following neutral lipid components: (1) cholesterol or a derivative thereof (2) a phospholipid; or (3) a mixture of a phospholipid and cholesterol or a derivative thereof. Examples of cholesterol derivatives include, but are not limited to, cholestanol, cholestanone, cholestenone, coprostanol, cholesteryl-2’-hydroxyethyl ether, cholesteryl-4’- hydroxybutyl ether, and mixtures thereof. The synthesis of cholesteryl-2’-hydroxyethyl ether is known to one skilled in the art and described in U.S. Patent Nos.8,058,069, 8,492,359, 8,822,668, 9,364,435, 9,504,651, and 11,141,378, all of which are hereby incorporated herein in their entireties for all purposes. Non-limiting examples of non-cationic lipids include phospholipids such as lecithin, phosphatidylethanolamine, lysolecithin, lysophosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, egg sphingomyelin (ESM), cephalin, cardiolipin, phosphatidic acid, cerebrosides, dicetylphosphate, distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), ioleoylphosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoylphosphatidylethanolamine (POPE), palmitoyloleyolphosphatidylglycerol (POPG), dioleoylphosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-l- carboxylate DOPE-mal), dipalmitoylphosphatidylethanolamine (DPPE), dimyristoylphosphatidylethanolamine (DMPE), distearoylphosphatidylethanolamine (DSPE), monomethylphosphatidylethanolamine, dimethylphosphatidylethanolamine, dielaidoylphosphatidylethanolamine (DEPE), stearoyloleoylphosphatidylethanolamine (SOPE), lysophosphatidylcholine, dilinoleoylphosphatidylcholine, and mixtures thereof. Other diacylphosphatidylcholine and diacylphosphatidylethanolamine phospholipids - 81 - 53224792.3 Attorney Docket No.0466483-7483WO1 can also be used. The acyl groups in these lipids can be, for example, acyl groups derived from fatty acids having C10-C24carbon chains, e.g., lauroyl, myristoyl, palmitoyl, stearoyl, or oleoyl. Additional examples of non-cationic lipids include sterols such as cholesterol and derivatives thereof such as cholestanol, cholestanone, cholestenone, coprostanol, cholesteryl- 2’-hydroxyethyl ether, cholesteryl-4’-hydroxybutyl ether, and mixtures thereof. In certain embodiments, the phospholipid is DPPC, DSPC, or mixtures thereof. Polymer-Conjugated Lipid(s) In the nucleic acid-lipid particles of the present disclosure, the conjugated lipid that inhibits aggregation of particles may comprise, e.g., one or more of the following: a polyethyleneglycol (PEG) lipid conjugate, a polyamide (ATTA)-lipid conjugate, a cationic- polymer-lipid conjugates (CPLs), or mixtures thereof. In some embodiments, the nucleic acid-lipid particles comprise either a PEG-lipid conjugate or an ATTA-lipid conjugate. PEG is a linear, water-soluble polymer of ethylene PEG repeating units with two terminal hydroxyl groups. PEGs are classified by their molecular weights; for example, PEG 2000 has an average molecular weight of about 2,000 daltons, and PEG 5000 has an average molecular weight of about 5,000 daltons. PEGs are commercially available from Sigma Chemical Co. and other companies and include, for example, the following: monomethoxypolyethylene glycol (MePEGOH), monomethoxypolyethylene glycolsuccinate (MePEGS), monomethoxypolyethylene glycolsuccinimidyl succinate (MePEG-S-NHS), monomethoxypolyethylene glycolamine (MePEG-NH2), monomethoxypolyethylene glycoltresylate (MePEG-TRES), and monomethoxypolyethylene glycolimidazolylcarbonyl (MePEG-IM). Other PEGs such as those described in U.S. Patent Nos.6,774,180 and 7,053,150 (e.g., mPEG (20 KDa) amine) are also useful for preparing the PEG-lipid conjugates of the present disclosure. The disclosures of these patents are herein incorporated by reference in their entirety for all purposes. In addition, monomethoxypolyethyleneglycolacetic acid (MePEG-CH2COOH) is particularly useful for preparing PEG-lipid conjugates including, e.g., PEG-DAA conjugates. In certain embodiments, the PEG-lipid conjugate or ATTA-lipid conjugate is used together with a CPL. The conjugated lipid that inhibits aggregation of particles may comprise a PEG-lipid including, e.g., a PEG-diacylglycerol (DAG), a PEG dialkyloxypropyl (DAA), a PEG-phospholipid, a PEG-ceramide (Cer), or mixtures thereof. The PEGDAA conjugate may be PEG-dilauryloxypropyl (C12), a PEG-dimyristyloxypropyl (C14), a PEG- dipalmityloxypropyl (C16), a PEG-distearyloxypropyl (C18), or mixtures thereof. - 82 - 53224792.3 Attorney Docket No.0466483-7483WO1 Additional PEG-lipid conjugates suitable for use in the disclosure include, but are not limited to, mPEG2000-l,2-diO-alkyl-sn3-carbomoylglyceride (PEG-C-DOMG). The synthesis of PEG-C-DOMG is described in PCT Application No. PCT / US08 / 88676, filed December 31, 2008, the disclosure of which is herein incorporated by reference in its entirety for all purposes. Yet additional PEG-lipid conjugates suitable for use in the disclosure include, without limitation, l-[8’-(l,2-dimyristoyl-3-propanoxy)-carboxamido-3’,6’- dioxaoctanyl] carbamoyl-methyl-poly(ethylene glycol) (2 KPEG-DMG). The synthesis of 2 KPEG-DMG is described in U.S. Patent No.7,404,969, the disclosure of which is herein incorporated by reference in its entirety for all purposes. The PEG moiety of the PEG-lipid conjugates described herein may comprise an average molecular weight ranging from about 550 daltons to about 10,000 daltons. In certain instances, the PEG moiety has an average molecular weight of from about 750 daltons to about 5,000 daltons (e.g., from about 1,000 daltons to about 5,000 daltons, from about 1,500 daltons to about 3,000 daltons, from about 750 daltons to about 3,000 daltons, from about 750 daltons to about 2,000 daltons, etc.). In some embodiments, the PEG moiety has an average molecular weight of about 2,000 daltons or about 750 daltons. In addition to the foregoing, it will be readily apparent to those of skill in the art that other hydrophilic polymers can be used in place of PEG. Examples of suitable polymers that can be used in place of PEG include, but are not limited to, polyvinylpyrrolidone, polymethyloxazoline, polyethyloxazoline, polyhydroxypropyl methacrylamide, polymethacrylamide and polydimethylacrylamide, polylactic acid, polyglycolic acid, and derivatized celluloses such as hydroxymethylcellulose or hydroxyethylcellulose. In addition to the foregoing components, the particles (e.g., LNP) of the present disclosure can further comprise cationic poly(ethylene glycol) (PEG) lipids or CPLs (e.g., Chen et al., Bioconj. Chem., 11:433-437 (2000)). Suitable SPLPs and SPLP-CPLs for use in the present disclosure, and methods of making and using SPLPs and SPLP-CPLs, are disclosed, e.g., in U.S. Patent No.6,852,334 and PCT Publication No. WO 00 / 62813, the disclosures of which are herein incorporated by reference in their entirety for all purposes. In certain instances, the conjugated lipid that inhibits aggregation of particles (e.g., PEG-lipid conjugate) may comprise from about 0.1 mol% to about 2 mol%, from about 0.5 mol% to about 2 mol%, from about 1 mol% to about 2 mol%, from about 0.6 mol% to about 1.9 mol%, from about 0.7 mol% to about 1.8 mol%, from about 0.8 mol% to about 1.7 mol%, from about 1 mol% to about 1.8 mol%, from about 1.2 mol% to about 1.8 mol%, from about 1.2 mol% to about 1.7 mol%, from about 1.3 mol% to about 1.6 mol%, from about 1.4 mol% - 83 - 53224792.3 Attorney Docket No.0466483-7483WO1 to about 1.5 mol%, or about 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, or 2 mol% (or any fraction thereof or range therein) of the total lipid present in the particle. In the lipid nanoparticles of the present disclosure, the active agent or therapeutic agent may be fully encapsulated within the lipid portion of the particle, thereby protecting the active agent or therapeutic agent from enzymatic degradation. In some embodiments, a nucleic acid-lipid particle comprising a nucleic acid such as a messenger RNA (i.e., mRNA) is fully encapsulated within the lipid portion of the particle, thereby protecting the nucleic acid from nuclease degradation. In certain instances, the nucleic acid in the nucleic acid-lipid particle is not substantially degraded after exposure of the particle to a nuclease at 37° C. for at least about 20, 30, 45, or 60 minutes. In certain other instances, the nucleic acid in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C. for at least about 30, 45, or 60 minutes or at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, or 36 hours. In other embodiments, the active agent or therapeutic agent (e.g., nucleic acid such as siRNA) is complexed with the lipid portion of the particle. One of the benefits of the formulations of the present disclosure is that the lipid particle compositions are substantially non-toxic to mammals such as humans. Lipid Nanoparticles (LNPs) Comprising Aryl-Alkyl Disulfide Ionizable Lipid Compounds In another aspect, the disclosure provides a lipid nanoparticle (LNP). In certain embodiments, the LNP comprises at least one ionizable lipid. In certain embodiments, the ionizable lipid is a compound of formula (I). In certain embodiments, the LNP comprises a neutral lipid. In certain embodiments, the LNP comprises at least one cholesterol lipid and / or a modified derivative thereof. In certain embodiments, the LNP comprises at least one polymer-conjugated lipid and / or a modified derivative thereof. In certain embodiments, the at least one ionizable lipid compound comprises less than about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, or 90 mol% of the LNP. In certain embodiments, the at least one ionizable lipid compound comprises about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, or 90 mol% of the LNP. In - 84 - 53224792.3 Attorney Docket No.0466483-7483WO1 certain embodiments, the at least one ionizable lipid compound comprises greater than about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, or 90 mol% of the LNP. In certain embodiments, the at least one ionizable lipid compound comprises less than about 35 mol% of the LNP. In certain embodiments, the at least one ionizable lipid compound comprises about 35 mol% of the LNP. In certain embodiments, the at least one ionizable lipid compound comprises greater than about 35 mol% of the LNP. In certain embodiments, the at least one neutral lipid comprises less than about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or about 40 mol% of the LNP. In certain embodiments, the at least one neutral lipid comprises about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or about 40 mol% of the LNP. In certain embodiments, the at least one neutral lipid comprises greater than about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or about 40 mol% of the LNP. In certain embodiments, the at least one neutral lipid comprises less than about 16 mol% of the LNP. In certain embodiments, the at least one neutral lipid comprises about 16 mol% of the LNP. In certain embodiments, the at least one neutral lipid comprises greater than about 16 mol% of the LNP. In certain embodiments, the neutral lipid comprises dioleoylphosphatidylethanolamine (DOPE). In certain embodiments, the neutral lipid consists essentially of dioleoylphosphatidylethanolamine (DOPE). In certain embodiments, the neutral lipid comprises distearoylphosphatidylcholine (DSPC). In certain embodiments, the neutral lipid consists essentially of distearoylphosphatidylcholine (DSPC). In certain embodiments, the neutral lipid comprises dioleoylphosphatidylcholine (DOPC). In certain embodiments, the neutral lipid consists essentially of dioleoylphosphatidylcholine (DOPC). In certain embodiments, the at least one cholesterol lipid and / or modified derivative thereof comprises less than about 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, or about 75 mol% of the LNP. In certain embodiments, the at least one cholesterol lipid and / or modified derivative thereof comprises about 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, - 85 - 53224792.3 Attorney Docket No.0466483-7483WO1 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, or about 75 mol% of the LNP. In certain embodiments, the at least one cholesterol lipid and / or modified derivative thereof comprises greater than about 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, or about 75 mol% of the LNP. In certain embodiments, the at least one cholesterol lipid and / or modified derivative thereof comprises less than about 46.5 mol% of the LNP. In certain embodiments, the at least one cholesterol lipid and / or modified derivative thereof comprises about 46.5 mol% of the LNP. In certain embodiments, the at least one cholesterol lipid and / or modified derivative thereof comprises greater than about 46.5 mol% of the LNP. In certain embodiments, the at least one cholesterol lipid and / or modified derivative thereof comprises or consists essentially of cholesterol. In certain embodiments, the at least one cholesterol lipid and / or modified derivative thereof consists essentially of cholesterol. In certain embodiments, the at least one polymer-conjugated lipid comprises less than about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7.6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.9, 10.0, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11.0, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12.0, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13.0, 13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14.0, 14.1, 14.2, 14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, or about 15 mol% of the LNP. In certain embodiments, the at least one polymer-conjugated lipid comprises about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7.6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.9, 10.0, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11.0, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12.0, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13.0, 13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14.0, 14.1, 14.2, 14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, or about 15 mol% of the LNP. In certain embodiments, the at least one polymer-conjugated lipid comprises greater than about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, - 86 - 53224792.3 Attorney Docket No.0466483-7483WO1 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7.6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.9, 10.0, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11.0, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12.0, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13.0, 13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14.0, 14.1, 14.2, 14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, or about 15 mol% of the LNP. In certain embodiments, the at least one polymer-conjugated lipid comprises less than about 2.5 mol%. In certain embodiments, the at least one polymer-conjugated lipid comprises about 2.5 mol%. In certain embodiments, the at least one polymer-conjugated lipid comprises greater than about 2.5 mol%. In certain embodiments, the at least one polymer-conjugated lipid comprises 1,2- dimyristoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethyleneglycol)-2000] (C14PEG2K). In certain embodiments, the at least one polymer-conjugated lipid consists essentially of 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy(polyethyleneglycol)-2000] (C14PEG2K). In certain embodiments, the LNP has a molar ratio of (a) : (b) : (c) : (d) of about 35:16:46.5:2.5. In certain embodiments, the LNP further comprises at least one cargo molecule. In certain embodiments, the at least one cargo molecule comprises or consists of a therapeutic cargo molecule. In certain embodiments, the cargo molecule is at least one selected from the group consisting of a nucleic acid, small molecule, protein, therapeutic agent, antibody, and any combinations thereof. In certain embodiments, the cargo molecule comprises a nucleic acid. In certain embodiments, the nucleic acid is DNA or RNA. In certain embodiments, the nucleic acid is selected from the group consisting of mRNA, cDNA, pDNA, microRNA, sgRNA, siRNA, modified RNA, antagomir, antisense molecule, and any combinations thereof. In certain embodiments, the mRNA encodes an enzyme. In certain embodiments, the mRNA encodes a receptor. In certain embodiments, the mRNA encodes an antigen binding domain. In certain embodiments, the mRNA encodes a clustered regularly interspaced short - 87 - 53224792.3 Attorney Docket No.0466483-7483WO1 palindrome repeats (CRISPR) associated protein. In certain embodiments, the CRISPR associated protein is Cas9. In certain embodiments, the nucleic acid cargo further comprises sgRNA. Lipid Nanoparticles (LNPs) Comprising Amidine-Crosslinker Containing Ionizable Lipid Compounds In another aspect, the disclosure provides a lipid nanoparticle (LNP). In certain embodiments, the LNP comprises at least one ionizable lipid. In certain embodiments, the ionizable lipid is a compound of formula (II). In certain embodiments, the LNP comprises a neutral lipid. In certain embodiments, the LNP comprises at least one cholesterol lipid and / or a modified derivative thereof. In certain embodiments, the LNP comprises at least one polymer-conjugated lipid and / or a modified derivative thereof. In certain embodiments, the at least one ionizable lipid compound comprises less than about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, or 90 mol% of the LNP. In certain embodiments, the at least one ionizable lipid compound comprises about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, or 90 mol% of the LNP. In certain embodiments, the at least one ionizable lipid compound comprises greater than about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, or 90 mol% of the LNP. In certain embodiments, the at least one ionizable lipid compound comprises less than about 57.1 mol% of the LNP. In certain embodiments, the at least one ionizable lipid compound comprises about 57.1 mol% of the LNP. In certain embodiments, the at least one ionizable lipid compound comprises greater than about 57.1 mol% of the LNP. In certain embodiments, the at least one ionizable lipid compound comprises less than about 50 mol% of the LNP. In certain embodiments, the at least one ionizable lipid compound comprises about 50 mol% of the LNP. In certain embodiments, the at least one ionizable lipid compound comprises greater than about 50 mol% of the LNP. - 88 - 53224792.3 Attorney Docket No.0466483-7483WO1 In certain embodiments, the at least ionizable lipid compound comprises or consists essentially of: (C12-2aN). In certain embodiments, the at least one neutral lipid comprises less than about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or about 40 mol% of the LNP. In certain embodiments, the at least one neutral lipid comprises about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or about 40 mol% of the LNP. In certain embodiments, the at least one neutral lipid comprises greater than about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or about 40 mol% of the LNP. In certain embodiments, the at least one neutral lipid comprises less than about 10 mol% of the LNP. In certain embodiments, the at least one neutral lipid comprises about 10 mol% of the LNP. In certain embodiments, the at least one neutral lipid comprises greater than about 10 mol% of the LNP. In certain embodiments, the at least one neutral lipid comprises less than about 10.2 mol% of the LNP. In certain embodiments, the at least one neutral lipid comprises about 10.2 mol% of the LNP. In certain embodiments, the at least one neutral lipid comprises greater than about 10.2 mol% of the LNP. In certain embodiments, the neutral lipid comprises dioleoylphosphatidylethanolamine (DOPE). In certain embodiments, the neutral lipid consists essentially of dioleoylphosphatidylethanolamine (DOPE). In certain embodiments, the neutral lipid comprises distearoylphosphatidylcholine (DSPC). In certain embodiments, the neutral lipid consists essentially of distearoylphosphatidylcholine (DSPC). In certain embodiments, the neutral lipid comprises dioleoylphosphatidylcholine (DOPC). In certain embodiments, the neutral lipid consists essentially of dioleoylphosphatidylcholine (DOPC). In certain embodiments, the at least one cholesterol lipid and / or modified derivative - 89 - 53224792.3 Attorney Docket No.0466483-7483WO1 thereof comprises less than about 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, or about 75 mol% of the LNP. In certain embodiments, the at least one cholesterol lipid and / or modified derivative thereof comprises about 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, or about 75 mol% of the LNP. In certain embodiments, the at least one cholesterol lipid and / or modified derivative thereof comprises greater than about 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, or about 75 mol% of the LNP. In certain embodiments, the at least one cholesterol lipid and / or modified derivative thereof comprises less than about 31.1 mol% of the LNP. In certain embodiments, the at least one cholesterol lipid and / or modified derivative thereof comprises about 31.1 mol% of the LNP. In certain embodiments, the at least one cholesterol lipid and / or modified derivative thereof comprises greater than about 31.1 mol% of the LNP. In certain embodiments, the at least one cholesterol lipid and / or modified derivative thereof comprises less than about 38.5 mol% of the LNP. In certain embodiments, the at least one cholesterol lipid and / or modified derivative thereof comprises about 38.5 mol% of the LNP. In certain embodiments, the at least one cholesterol lipid and / or modified derivative thereof comprises greater than about 38.5 mol% of the LNP. In certain embodiments, the at least one cholesterol lipid and / or modified derivative thereof comprises or consists essentially of cholesterol. In certain embodiments, the at least one cholesterol lipid and / or modified derivative thereof consists essentially of cholesterol. In certain embodiments, the at least one polymer-conjugated lipid comprises less than about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7.6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.9, 10.0, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11.0, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12.0, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13.0, 13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14.0, 14.1, 14.2, 14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, or about 15 mol% of the LNP. In certain embodiments, the at least one polymer-conjugated lipid comprises about - 90 - 53224792.3 Attorney Docket No.0466483-7483WO1 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7.6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.9, 10.0, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11.0, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12.0, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13.0, 13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14.0, 14.1, 14.2, 14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, or about 15 mol% of the LNP. In certain embodiments, the at least one polymer-conjugated lipid comprises greater than about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7.6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.9, 10.0, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11.0, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12.0, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13.0, 13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14.0, 14.1, 14.2, 14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, or about 15 mol% of the LNP. In certain embodiments, the at least one polymer-conjugated lipid comprises less than about 1.7 mol%. In certain embodiments, the at least one polymer-conjugated lipid comprises about 1.7 mol%. In certain embodiments, the at least one polymer-conjugated lipid comprises greater than about 1.7 mol%. In certain embodiments, the at least one polymer-conjugated lipid comprises less than about 1.5 mol%. In certain embodiments, the at least one polymer-conjugated lipid comprises about 1.5 mol%. In certain embodiments, the at least one polymer-conjugated lipid comprises greater than about 1.5 mol%. In certain embodiments, the at least one polymer-conjugated lipid comprises 1,2- dimyristoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethyleneglycol)-2000] (C14PEG2K). In certain embodiments, the at least one polymer-conjugated lipid consists essentially of 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy(polyethyleneglycol)-2000] (C14PEG2K). In certain embodiments, the LNP has a molar ratio of (a) : (b) : (c) : (d) of about 57.1 : 10.2 : 31.1 : 1.7. In certain embodiments, the LNP has a molar ratio of (a) : (b) : (c) : (d) of about 50 : 10 : 38.5 : 1.5. - 91 - 53224792.3 Attorney Docket No.0466483-7483WO1 In certain embodiments, the LNP further comprises at least one cargo molecule. In certain embodiments, the at least one cargo molecule comprises or consists of a therapeutic cargo molecule. In certain embodiments, the cargo molecule is at least one selected from the group consisting of a nucleic acid, small molecule, protein, therapeutic agent, antibody, and any combinations thereof. In certain embodiments, the cargo molecule comprises a nucleic acid. In certain embodiments, the nucleic acid is DNA or RNA. In certain embodiments, the nucleic acid is selected from the group consisting of mRNA, cDNA, pDNA, microRNA, sgRNA, siRNA, modified RNA, antagomir, antisense molecule, and any combinations thereof. In certain embodiments, the mRNA encodes an enzyme, receptor, or antigen binding domain. In certain embodiments, the mRNA encodes a clustered regularly interspaced short palindrome repeats (CRISPR) associated protein. In certain embodiments, the CRISPR associated protein is Cas9. In certain embodiments, the nucleic acid cargo further comprises sgRNA. Methods of Using Lipid Nanoparticles (LNPs) Comprising Aryl-Alkyl Disulfide Ionizable Lipid Compounds In one aspect, the disclosure provides a method of treating, preventing, and / or ameliorating a disease or infection in a subject. In certain embodiments, the method comprises administering to the subject at least one lipid nanoparticle (LNP) of the disclosure or a pharmaceutical composition thereof. In certain embodiments, the disease is cancer. In certain embodiments, the disease is an immunomodulatory disease. In certain embodiments, the disease is a lymphatic disease. In certain embodiments, the infection is a viral infection. In certain embodiments, the disclosure provides a method of delivering at least one therapeutic cargo molecule to a lymph node of a subject. In certain embodiments, the method comprises administering to the subject at least one lipid nanoparticle (LNP) of the disclosure or a pharmaceutical composition thereof. In certain embodiments, the lymph node is a pelvic lymph node. In certain embodiments, the pelvic lymph node is an inguinal lymph node. In certain embodiments, the pelvic lymph node is an iliac lymph node. - 92 - 53224792.3 Attorney Docket No.0466483-7483WO1 In certain embodiments, the subject is a mammal. In certain embodiments, the mammal is a human. Methods of Using Lipid Nanoparticles (LNPs) Comprising Amidine-Crosslinker Containing Ionizable Lipid Compounds In one aspect, the disclosure provides a method of treating, preventing, and / or ameliorating a disease in a subject. In certain embodiments, the method comprises administering to the subject at least one lipid nanoparticle (LNP) of the disclosure or a pharmaceutical composition of the disclosure. In certain embodiments, the disease is at least one selected from the group consisting of cancer, immune-mediated diseases, cardiovascular disease, and a metabolic disease. In certain embodiments, delivery of the at least one therapeutic cargo molecule is selective for the spleen. In certain embodiments, the at least one therapeutic cargo is not substantially delivered to the liver. In certain embodiments, the at least one therapeutic cargo is not substantially delivered to the heart. In certain embodiments, the at least one therapeutic cargo is not substantially delivered to the lungs. In certain embodiments, the at least one therapeutic cargo is not substantially delivered to the kidney. In certain embodiments, a greater proportion of the at least one therapeutic cargo molecule is delivered to the spleen than the liver. In certain embodiments, a greater proportion of the at least one therapeutic cargo molecule is delivered to the spleen than the heart. In certain embodiments, a greater proportion of the at least one therapeutic cargo molecule is delivered to the spleen than the lungs. In certain embodiments, a greater proportion of the at least one therapeutic cargo molecule is delivered to the spleen than the kidney. In certain embodiments, a greater proportion of the at least one therapeutic cargo molecule is delivered to the spleen than the sum of therapeutic cargo delivered to the liver, heart, lungs, and kidney. In certain embodiments, the at least one therapeutic cargo molecule is delivered to a splenic red pulp macrophage. In certain embodiments, the therapeutic cargo molecule is at least one selected from the group consisting of a nucleic acid, small molecule, protein, therapeutic agent, antibody, and any combinations thereof. In certain embodiments, the therapeutic cargo molecule comprises a nucleic acid. In certain embodiments, the nucleic acid is DNA or RNA. In certain embodiments, the nucleic acid is selected from the group consisting of mRNA, cDNA, pDNA, microRNA, sgRNA, siRNA, modified RNA, antagomir, antisense - 93 - 53224792.3 Attorney Docket No.0466483-7483WO1 molecule, and any combinations thereof. In another aspect, the disclosure provides a method of delivering at least one therapeutic cargo molecule to a dendritic cell of a subject. In certain embodiments, the method comprises administering to the subject at least one lipid nanoparticle (LNP) of the disclosure or a pharmaceutical composition of the disclosure. In certain embodiments, the therapeutic cargo molecule is mRNA. In certain embodiments, the LNP stimulates glycolysis in the dendritic cell. In certain embodiments, the LNP facilitates endosomal escape. In certain embodiments, the subject is a mammal. In certain embodiments, the mammal is a human. Pharmaceutical Compositions In another aspect, the present disclosure provides a pharmaceutical composition comprising the lipid nanoparticle (LNP) of the present disclosure and at least one pharmaceutically acceptable carrier. In certain embodiments, the composition further comprises at least one adjuvant. In certain embodiments, the composition is a vaccine. In certain embodiments, the composition of the disclosure is formulated for use as a vaccine. In certain embodiments, the composition further comprises at least one adjuvant. Such a pharmaceutical composition may consist of at least one composition of the invention, in a form suitable for administration to a subject, or the pharmaceutical composition may comprise at least one composition, and one or more pharmaceutically acceptable carriers, one or more additional ingredients, or any combinations of these. At least one composition of the invention may be present in the pharmaceutical composition in the form of a physiologically acceptable salt, such as in combination with a physiologically acceptable cation or anion, as is well known in the art. In certain embodiments, the pharmaceutical compositions useful for practicing the method of the invention may be administered to deliver a dose of between 1 ng / kg / day and 100 mg / kg / day. In other embodiments, the pharmaceutical compositions useful for practicing the invention may be administered to deliver a dose of between 1 ng / kg / day and 1,000 mg / kg / day. The relative amounts of the active ingredient, the pharmaceutically acceptable carrier, and any additional ingredients in a pharmaceutical composition of the invention will vary, depending upon the identity, size, and condition of the subject treated and further depending upon the route by which the composition is to be administered. By way of example, the - 94 - 53224792.3 Attorney Docket No.0466483-7483WO1 composition may comprise between 0.1% and 100% (w / w) active ingredient. Pharmaceutical compositions that are useful in the methods of the invention may be suitably developed for nasal, inhalational, oral, rectal, vaginal, pleural, peritoneal, parenteral, topical, transdermal, pulmonary, intranasal, buccal, ophthalmic, epidural, intrathecal, intravenous, or another route of administration. A composition useful within the methods of the invention may be directly administered to the brain, the brainstem, or any other part of the central nervous system of a mammal or bird. Other contemplated formulations include projected nanoparticles, microspheres, liposomal preparations, coated particles, polymer conjugates, resealed erythrocytes containing the active ingredient, and immunologically- based formulations. In certain embodiments, the compositions of the invention are part of a pharmaceutical matrix, which allows for manipulation of insoluble materials and improvement of the bioavailability thereof, development of controlled or sustained release products, and generation of homogeneous compositions. By way of example, a pharmaceutical matrix may be prepared using hot melt extrusion, solid solutions, solid dispersions, size reduction technologies, molecular complexes (e.g., cyclodextrins, and others), microparticulate, and particle and formulation coating processes. Amorphous or crystalline phases may be used in such processes. The route(s) of administration will be readily apparent to the skilled artisan and will depend upon any number of factors including the type and severity of the disease being treated, the type and age of the veterinary or human patient being treated, and the like. The formulations of the pharmaceutical compositions described herein may be prepared by any method known or hereafter developed in the art of pharmacology and pharmaceutics. In general, such preparatory methods include the step of bringing the active ingredient into association with a carrier or one or more other accessory ingredients, and then, if necessary or desirable, shaping or packaging the product into a desired single-dose or multi-dose unit. As used herein, a “unit dose” is a discrete amount of the pharmaceutical composition comprising a predetermined amount of the active ingredient. The amount of the active ingredient is generally equal to the dosage of the active ingredient that would be administered to a subject or a convenient fraction of such a dosage such as, for example, one-half or one- third of such a dosage. The unit dosage form may be for a single daily dose or one of multiple daily doses (e.g., about 1 to 4 or more times per day). When multiple daily doses are used, the unit dosage form may be the same or different for each dose. - 95 - 53224792.3 Attorney Docket No.0466483-7483WO1 Although the descriptions of pharmaceutical compositions provided herein are principally directed to pharmaceutical compositions suitable for ethical administration to humans, it will be understood by the skilled artisan that such compositions are generally suitable for administration to animals of all sorts. Modification of pharmaceutical compositions suitable for administration to humans in order to render the compositions suitable for administration to various animals is well understood, and the ordinarily skilled veterinary pharmacologist can design and perform such modification with merely ordinary, if any, experimentation. Subjects to which administration of the pharmaceutical compositions of the invention is contemplated include, but are not limited to, humans and other primates, mammals including commercially relevant mammals such as cattle, pigs, horses, sheep, cats, and dogs. In certain embodiments, the compositions of the invention are formulated using one or more pharmaceutically acceptable excipients or carriers. In certain embodiments, the pharmaceutical compositions of the invention comprise a therapeutically effective amount of at least one compound of the invention and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers, which are useful, include, but are not limited to, glycerol, water, saline, ethanol, recombinant human albumin (e.g., RECOMBUMIN®), solubilized gelatins (e.g., GELOFUSINE®), and other pharmaceutically acceptable salt solutions such as phosphates and salts of organic acids. Examples of these and other pharmaceutically acceptable carriers are described in Remington’s Pharmaceutical Sciences (1991, Mack Publication Co., New Jersey). The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), recombinant human albumin, solubilized gelatins, suitable mixtures thereof, and vegetable oils. The proper fluidity may be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms may be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, isotonic agents, for example, sugars, sodium chloride, or polyalcohols such as mannitol and sorbitol, are included in the composition. Prolonged absorption of the injectable compositions may be brought about by including in the composition an agent that delays absorption, for example, aluminum monostearate or gelatin. Formulations may be employed in admixtures with conventional excipients, i.e., - 96 - 53224792.3 Attorney Docket No.0466483-7483WO1 pharmaceutically acceptable organic or inorganic carrier substances suitable for oral, parenteral, nasal, inhalational, intravenous, subcutaneous, transdermal enteral, or any other suitable mode of administration, known to the art. The pharmaceutical preparations may be sterilized and if desired mixed with auxiliary agents, e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure buffers, coloring, flavoring, and / or fragrance-conferring substances and the like. They may also be combined where desired with other active agents, e.g., other analgesic, anxiolytics or hypnotic agents. As used herein, “additional ingredients” include, but are not limited to, one or more ingredients that may be used as a pharmaceutical carrier. The composition of the invention may comprise a preservative from about 0.005% to 2.0% by total weight of the composition. The preservative is used to prevent spoilage in the case of exposure to contaminants in the environment. Examples of preservatives useful in accordance with the invention include but are not limited to those selected from the group consisting of benzyl alcohol, sorbic acid, parabens, imidurea and any combinations thereof. One such preservative is a combination of about 0.5% to 2.0% benzyl alcohol and 0.05-0.5% sorbic acid. The composition may include an antioxidant and a chelating agent that inhibit the degradation of the compound. Antioxidants for some compounds are BHT, BHA, alpha- tocopherol and ascorbic acid in the exemplary range of about 0.01% to 0.3%, or BHT in the range of 0.03% to 0.1% by weight by total weight of the composition. The chelating agent may be present in an amount of from 0.01% to 0.5% by weight by total weight of the composition. Exemplary chelating agents include edetate salts (e.g. disodium edetate) and citric acid in the weight range of about 0.01% to 0.20%, or in the range of 0.02% to 0.10% by weight by total weight of the composition. The chelating agent is useful for chelating metal ions in the composition that may be detrimental to the shelf life of the formulation. While BHT and disodium edetate are exemplary antioxidant and chelating agent, respectively, for some compounds, other suitable and equivalent antioxidants and chelating agents may be substituted therefore as would be known to those skilled in the art. Liquid suspensions may be prepared using conventional methods to achieve suspension of the active ingredient in an aqueous or oily vehicle. Aqueous vehicles include, for example, water, and isotonic saline. Oily vehicles include, for example, almond oil, oily esters, ethyl alcohol, vegetable oils such as arachis, olive, sesame, or coconut oil, fractionated vegetable oils, and mineral oils such as liquid paraffin. Liquid suspensions may further comprise one or more additional ingredients including, but not limited to, suspending agents, - 97 - 53224792.3 Attorney Docket No.0466483-7483WO1 dispersing or wetting agents, emulsifying agents, demulcents, preservatives, buffers, salts, flavorings, coloring agents, and sweetening agents. Oily suspensions may further comprise a thickening agent. Known suspending agents include, but are not limited to, sorbitol syrup, hydrogenated edible fats, sodium alginate, polyvinylpyrrolidone, gum tragacanth, gum acacia, and cellulose derivatives such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethyl cellulose. Known dispersing or wetting agents include, but are not limited to, naturally-occurring phosphatides such as lecithin, condensation products of an alkylene oxide with a fatty acid, with a long chain aliphatic alcohol, with a partial ester derived from a fatty acid and a hexitol, or with a partial ester derived from a fatty acid and a hexitol anhydride (e.g., polyoxyethylene stearate, heptadecaethyleneoxycetanol, polyoxyethylene sorbitol monooleate, and polyoxyethylene sorbitan monooleate, respectively). Known emulsifying agents include, but are not limited to, lecithin, acacia, and ionic or non-ionic surfactants. Known preservatives include, but are not limited to, methyl, ethyl, or n-propyl para-hydroxybenzoates, ascorbic acid, and sorbic acid. Known sweetening agents include, for example, glycerol, propylene glycol, sorbitol, sucrose, and saccharin. Liquid solutions of the active ingredient in aqueous or oily solvents may be prepared in substantially the same manner as liquid suspensions, the primary difference being that the active ingredient is dissolved, rather than suspended in the solvent. As used herein, an “oily” liquid is one which comprises a carbon-containing liquid molecule and which exhibits a less polar character than water. Liquid solutions of the pharmaceutical composition of the invention may comprise each of the components described with regard to liquid suspensions, it being understood that suspending agents will not necessarily aid dissolution of the active ingredient in the solvent. Aqueous solvents include, for example, water, and isotonic saline. Oily solvents include, for example, almond oil, oily esters, ethyl alcohol, vegetable oils such as arachis, olive, sesame, or coconut oil, fractionated vegetable oils, and mineral oils such as liquid paraffin. A pharmaceutical composition of the invention may also be prepared, packaged, or sold in the form of oil-in-water emulsion or a water-in-oil emulsion. The oily phase may be a vegetable oil such as olive or arachis oil, a mineral oil such as liquid paraffin, or a combination of these. Such compositions may further comprise one or more emulsifying agents such as naturally occurring gums such as gum acacia or gum tragacanth, naturally- occurring phosphatides such as soybean or lecithin phosphatide, esters or partial esters derived from combinations of fatty acids and hexitol anhydrides such as sorbitan monooleate, and condensation products of such partial esters with ethylene oxide such as polyoxyethylene - 98 - 53224792.3 Attorney Docket No.0466483-7483WO1 sorbitan monooleate. These emulsions may also contain additional ingredients including, for example, sweetening or flavoring agents. Methods for impregnating or coating a material with a chemical composition are known in the art, and include, but are not limited to methods of depositing or binding a chemical composition onto a surface, methods of incorporating a chemical composition into the structure of a material during the synthesis of the material (i.e., such as with a physiologically degradable material), and methods of absorbing an aqueous or oily solution or suspension into an absorbent material, with or without subsequent drying. Methods for mixing components include physical milling, the use of pellets in solid and suspension formulations and mixing in a transdermal patch, as known to those skilled in the art. Administration / Dosing The regimen of administration may affect what constitutes an effective amount. The therapeutic formulations may be administered to the patient either prior to or after the onset of a disease or disorder. Further, several divided dosages, as well as staggered dosages may be administered daily or sequentially, or the dose may be continuously infused, or may be a bolus injection. Further, the dosages of the therapeutic formulations may be proportionally increased or decreased as indicated by the exigencies of the therapeutic or prophylactic situation. Administration of the compositions of the present disclosure to a patient, such as a mammal, such as a human, may be carried out using known procedures, at dosages and for periods of time effective to treat a disease or disorder contemplated herein. An effective amount of therapeutic (i.e., composition) necessary to achieve a therapeutic effect may vary according to factors such as the activity of the particular therapeutic employed; the time of administration; the rate of excretion of the composition; the duration of the treatment; other drugs, compounds or materials used in combination with the composition; the state of the disease or disorder, age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well-known in the medical arts. Dosage regimens may be adjusted to provide the optimum therapeutic response. For example, several divided doses may be administered daily or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation. A non-limiting example of an effective dose range for a therapeutic composition of the disclosure is from about 0.01 mg / kg to 100 mg / kg of body weight / per day of active agent (i.e., nucleic acid). One of ordinary skill in the art would be able to study the relevant factors and make the determination regarding the effective - 99 - 53224792.3 Attorney Docket No.0466483-7483WO1 amount of the therapeutic composition without undue experimentation. The composition may be administered to an animal as frequently as several times daily, or it may be administered less frequently, such as once a day, once a week, once every two weeks, once a month, or even less frequently, such as once every several months or even once a year or less. It is understood that the amount of composition dosed per day may be administered, in non-limiting examples, every day, every other day, every 2 days, every 3 days, every 4 days, or every 5 days. For example, with every other day administration, a 5 mg per day dose may be initiated on Monday with a first subsequent 5 mg per day dose administered on Wednesday, a second subsequent 5 mg per day dose administered on Friday, and so on. The frequency of the dose is readily apparent to the skilled artisan and depends upon a number of factors, such as, but not limited to, type and severity of the disease being treated, and type and age of the animal. Actual dosage levels of the active ingredients in the pharmaceutical compositions of this disclosure may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient. A medical doctor, e.g., physician or veterinarian, having ordinary skill in the art may readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the compounds of the disclosure employed in the pharmaceutical composition at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. In particular embodiments, it is especially advantageous to formulate the compound in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the patients to be treated; each unit containing a predetermined quantity of therapeutic composition to produce the desired therapeutic effect in association with the required pharmaceutical vehicle. The dosage unit forms of the disclosure are dictated by and directly dependent on (a) the unique characteristics of the therapeutic composition and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding / formulating such a therapeutic composition for the treatment of a disease or disorder in a patient. In certain embodiments, the compositions of the disclosure are administered to the patient in dosages that range from one to five times per day or more. In other embodiments, the compositions of the disclosure are administered to the patient in range of dosages that - 100 - 53224792.3 Attorney Docket No.0466483-7483WO1 include, but are not limited to, once every day, every two days, every three days to once a week, and once every two weeks. It will be readily apparent to one skilled in the art that the frequency of administration of the various combination compositions of the disclosure will vary from subject to subject depending on many factors including, but not limited to, age, disease or disorder to be treated, gender, overall health, and other factors. Thus, the disclosure should not be construed to be limited to any particular dosage regime and the precise dosage and composition to be administered to any patient will be determined by the attending physician taking all other factors about the patient into account. The amount of active agent of the composition(s) of the disclosure for administration may be in the range of from about 1 µg to about 7,500 mg, about 20 µg to about 7,000 mg, about 40 µg to about 6,500 mg, about 80 µ g to about 6,000 mg, about 100 µ g to about 5,500 mg, about 200 µ g to about 5,000 mg, about 400 µ g to about 4,000 mg, about 800 µ g to about 3,000 mg, about 1 mg to about 2,500 mg, about 2 mg to about 2,000 mg, about 5 mg to about 1,000 mg, about 10 mg to about 750 mg, about 20 mg to about 600 mg, about 30 mg to about 500 mg, about 40 mg to about 400 mg, about 50 mg to about 300 mg, about 60 mg to about 250 mg, about 70 mg to about 200 mg, about 80 mg to about 150 mg, and any and all whole or partial increments there-in-between. In some embodiments, the dose of active agent (i.e., nucleic acid) present in the composition of the disclosure is from about 0.5 µg and about 5,000 mg. In some embodiments, a dose of active agent present in the composition of the disclosure used in compositions described herein is less than about 5,000 mg, or less than about 4,000 mg, or less than about 3,000 mg, or less than about 2,000 mg, or less than about 1,000 mg, or less than about 800 mg, or less than about 600 mg, or less than about 500 mg, or less than about 200 mg, or less than about 50 mg. Similarly, in some embodiments, a dose of a second compound as described herein is less than about 1,000 mg, or less than about 800 mg, or less than about 600 mg, or less than about 500 mg, or less than about 400 mg, or less than about 300 mg, or less than about 200 mg, or less than about 100 mg, or less than about 50 mg, or less than about 40 mg, or less than about 30 mg, or less than about 25 mg, or less than about 20 mg, or less than about 15 mg, or less than about 10 mg, or less than about 5 mg, or less than about 2 mg, or less than about 1 mg, or less than about 0.5 mg, and any and all whole or partial increments thereof. In certain embodiments, the present disclosure is directed to a packaged pharmaceutical composition comprising a container holding a therapeutically effective amount of the composition of the disclosure, alone or in combination with a second - 101 - 53224792.3 Attorney Docket No.0466483-7483WO1 pharmaceutical agent; and instructions for using the compound to treat, prevent, or reduce one or more symptoms of a disease or disorder in a patient. The term “container” includes any receptacle for holding the pharmaceutical composition or for managing stability or water uptake. For example, in certain embodiments, the container is the packaging that contains the pharmaceutical composition, such as liquid (solution and suspension), semisolid, lyophilized solid, solution and powder or lyophilized formulation present in dual chambers. In other embodiments, the container is not the packaging that contains the pharmaceutical composition, i.e., the container is a receptacle, such as a box or vial that contains the packaged pharmaceutical composition or unpackaged pharmaceutical composition and the instructions for use of the pharmaceutical composition. Moreover, packaging techniques are well known in the art. It should be understood that the instructions for use of the pharmaceutical composition may be contained on the packaging containing the pharmaceutical composition, and as such the instructions form an increased functional relationship to the packaged product. However, it should be understood that the instructions may contain information pertaining to the compound’s ability to perform its intended function, e.g., treating, preventing, or reducing a disease or disorder in a patient. Administration Routes of administration of any of the compositions of the disclosure include inhalational, oral, nasal, rectal, parenteral, sublingual, transdermal, transmucosal (e.g., sublingual, lingual, (trans)buccal, (trans)urethral, vaginal (e.g., trans- and perivaginally), (intra)nasal, and (trans)rectal), intravesical, intrapulmonary, intraduodenal, intragastrical, intrathecal, epidural, intrapleural, intraperitoneal, subcutaneous, intramuscular, intradermal, intra-arterial, intravenous, intrabronchial, inhalation, and topical administration. Suitable compositions and dosage forms include, for example, tablets, capsules, caplets, pills, gel caps, troches, emulsions, dispersions, suspensions, solutions, syrups, granules, beads, transdermal patches, gels, powders, pellets, magmas, lozenges, creams, pastes, plasters, lotions, discs, suppositories, liquid sprays for nasal or oral administration, dry powder or aerosolized formulations for inhalation, compositions and formulations for intravesical administration and the like. It should be understood that the formulations and compositions that would be useful in the present disclosure are not limited to the particular formulations and compositions that are described herein. Parenteral Administration - 102 - 53224792.3 Attorney Docket No.0466483-7483WO1 As used herein, “parenteral administration” of a pharmaceutical composition includes any route of administration characterized by physical breaching of a tissue of a subject and administration of the pharmaceutical composition through the breach in the tissue. Parenteral administration thus includes, but is not limited to, administration of a pharmaceutical composition by injection of the composition, by application of the composition through a surgical incision, by application of the composition through a tissue-penetrating non-surgical wound, and the like. In particular, parenteral administration is contemplated to include, but is not limited to, subcutaneous, intravenous, intraperitoneal, intramuscular, intrasternal injection, and kidney dialytic infusion techniques. Formulations of a pharmaceutical composition suitable for parenteral administration comprise the active ingredient combined with a pharmaceutically acceptable carrier, such as sterile water or sterile isotonic saline. Such formulations may be prepared, packaged, or sold in a form suitable for bolus administration or for continuous administration. Injectable formulations may be prepared, packaged, or sold in unit dosage form, such as in ampules or in multidose containers containing a preservative. Injectable formulations may also be prepared, packaged, or sold in devices such as patient-controlled analgesia (PCA) devices. Formulations for parenteral administration include, but are not limited to, suspensions, solutions, emulsions in oily or aqueous vehicles, pastes, and implantable sustained-release or biodegradable formulations. Such formulations may further comprise one or more additional ingredients including, but not limited to, suspending, stabilizing, or dispersing agents. In certain embodiments of a formulation for parenteral administration, the active ingredient is provided in dry (i.e., powder or granular) form for reconstitution with a suitable vehicle (e.g., sterile pyrogen-free water) prior to parenteral administration of the reconstituted composition. The pharmaceutical compositions may be prepared, packaged, or sold in the form of a sterile injectable aqueous or oily suspension or solution. This suspension or solution may be formulated according to the known art, and may comprise, in addition to the active ingredient, additional ingredients such as the dispersing agents, wetting agents, or suspending agents described herein. Such sterile injectable formulations may be prepared using a non- toxic parenterally acceptable diluent or solvent, such as water or 1,3-butanediol, for example. Other acceptable diluents and solvents include, but are not limited to, Ringer’s solution, isotonic sodium chloride solution, and fixed oils such as synthetic mono- or di-glycerides. Other parentally-administrable formulations which are useful include those which comprise the active ingredient in microcrystalline form in a recombinant human albumin, a fluidized gelatin, in a liposomal preparation, or as a component of a biodegradable polymer system. - 103 - 53224792.3 Attorney Docket No.0466483-7483WO1 Compositions for sustained release or implantation may comprise pharmaceutically acceptable polymeric or hydrophobic materials such as an emulsion, an ion exchange resin, a sparingly soluble polymer, or a sparingly soluble salt. EXAMPLES Various embodiments of the present application can be better understood by reference to the following Examples which are offered by way of illustration. The scope of the present application is not limited to the Examples given herein. Example 1: Design and Synthesis of Aromatic Ionizable Lipids of the Disclosure In one aspect, it was hypothesized that the use of ionizable lipids comprising aromatic and / or disulfide linking groups would permit exploration of structure-function relationships upon formulation thereof to lipid nanoparticles (LNPs). In certain embodiments, disulfide bonds increase biodegradability. In certain embodiments, π-stacking interactions may aid in mRNA encapsulation. Further, the use of disulfide-substituted aryl moieties permits exploration of the relationship between steric arrangement and in vivo properties of the resultant LNPs. The ionizable lipid compounds of the disclosure are referred to herein utilizing C[n][regiochemistry]PhE-[core] nomenclature, wherein “[n]” refers to the tail length (i.e., number of carbon atoms in alkyl substituent of the disulfide moiety), “[regiochemistry],” as indicated by “o,” “m,” and “p,” for ortho-, meta-, and para-substitution, respectively, refers to the relative connectivity with respect to the disulfide moiety and the linkage to the amine “core” on the aromatic ring to which both are attached (e.g., 1,2-substitution is ortho, 1,3- substitution is meta, and 1,4-substitution is para), and “[core]” refers to a numerical identifier for the corresponding amine moiety utilized to prepare the ionizable lipid. Non-limiting, exemplary nomenclature utilizing amine core 383 is provided in Table 1. Table 1. Exemplary nomenclature for compounds of the disclosure using representative core 383 Tail Length Ortho Disulfide Meta Disulfide Para Disulfide In certain embodiments, ionizable lipid compounds of the disclosure were prepared - 104 - 53224792.3 Attorney Docket No.0466483-7483WO1 using amine cores 200, 318, and 383 (i.e., [core] is 200, 318, or 383), the chemical structures of which are provided in Table 2. Table 2. Exemplary amine cores of the disclosure Core No. Compound 200 , to the non-limiting, exemplary synthetic methods depicted in Scheme 1, wherein exemplary amine core 318 is depicted for illustrative purposes. It is understood by those of ordinary skill in the art that the methods of preparing ionizable lipids described in the disclosure are not limited to the embodiments represented in Scheme 1. In certain embodiments, ionizable lipids of the disclosure were prepared according to the method depicted in Scheme 1 utilizing an alkanethiol (e.g., hexane-1-thiol, octane-1-thiol, decane-1-thiol, or dodecane-1-thiol) a thiophenol (e.g., 2-mercaptophenol, 3-mercaptophenol, or 4-mercaptophenol), and a suitable amine core (e.g., 200, 318, or 383). Exemplary ionizable lipids of the disclosure prepared according to Scheme 1 are provided herein (Table 3). - 105 - 53224792.3 Attorney Docket No.0466483-7483WO1 Table 3. Exemplary ionizable lipid compounds of the disclosure Name Compound - 106 - 53224792.3 Attorney Docket No.0466483-7483WO1 (decyldisulfaneyl)phenoxy)propan-2-ol) - 107 - 53224792.3 Attorney Docket No.0466483-7483WO1 yl)ethyl)azanediyl)bis(1-(3-(octyldisulfaneyl)phenoxy)propan-2-ol) - 108 - 53224792.3 Attorney Docket No.0466483-7483WO1 (hexyldisulfaneyl)phenoxy)propan-2-ol) S S - 109 - 53224792.3 Attorney Docket No.0466483-7483WO1 3,3'-((2-(4-(2-((2-(bis(3-(4-(dodecyldisulfaneyl)phenoxy)-2- hydroxypropyl)amino)ethyl)(3-(4-(dodecyldisulfaneyl)phenoxy)-2- hdrox ro l)amino)ethl)ierazin-1-l)ethl)azanedil)bis(1-(4- 1- - 110 - 53224792.3 Attorney Docket No.0466483-7483WO1 1- - 111 - 53224792.3 Attorney Docket No.0466483-7483WO1 OH O 1- - 112 - 53224792.3 Attorney Docket No.0466483-7483WO1 - 113 - 53224792.3 Attorney Docket No.0466483-7483WO1 ) - 114 - 53224792.3 Attorney Docket No.0466483-7483WO1 ) - 115 - 53224792.3 Attorney Docket No.0466483-7483WO1 ) Example 2: Lipid Nanoparticle Formulation and Characterization Exemplary lipid nanoparticles (LNPs) were prepared utilizing certain exemplary ionizable lipids of the disclosure and one or more neutral lipid(s), cholesterol lipid(s) and / or modified derivatives thereof, and polymer-conjugated lipid(s) and / or modified derivatives - 116 - 53224792.3 Attorney Docket No.0466483-7483WO1 thereof. Exemplary LNPs of the disclosure were formulated according to methods known to those of ordinary skill in the art. In certain non-limiting embodiments, LNPs of the disclosure were prepared as follows: An ethanol phase containing all lipids and an aqueous phase containing mRNA, where applicable, were mixed using a microfluidic device to formulate LNPs. The ethanol phase contained the ionizable lipid or normal ionizable lipid, 1,2-dioleoyl- sn-glycero-3-phosphoethanolamine (DOPE), cholesterol, and 1,2-dimyristoyl-sn-glycero-3- phosphoethanolamine-N- [methoxy(polyethyleneglycol)-2000] (C14PEG2K) with a fixed molar ratio of 35%, 16%, 46.5% and 2.5%, respectively. Where applicable, the aqueous phase comprised mRNA dissolved in 10 mM citrate buffer. The ethanol and aqueous phases were mixed at a flow rate of 1.8 ml / min and 0.6 ml / min (3:1) using Pump33DS syringe pumps. LNPs were dialyzed in 1x PBS using a microdialysis cassette (20,000 MWCO, Thermo Fisher Scientific, Waltham, MA) for 2 h and then filtered through a 0.22 μm filter. Certain exemplary LNPs of the disclosure comprising aromatic ionizable lipids of the disclosure were characterized with respect to encapsulation efficiency (FIG.1A), charge (FIG.1B), hydrodynamic diameter (FIG.1C), and polydispersity index (FIG.1D). Example 3: In vitro transfection and toxicity studies utilizing LNPs of the disclosure Certain exemplary LNPs of the disclosure comprising aromatic ionizable lipids of the disclosure and luciferase mRNA were analyzed for transfection efficacy (FIG.2A) and toxicity (FIG.2B) in vitro using HepG2 cells. These studies demonstrated that the LNPs of the disclosure have minimal toxicity and provide promising transfection efficacy. Example 4: In vivo studies utilizing LNPs of the disclosure Distribution of exemplary LNPs of the disclosure comprising aromatic ionizable lipids of the disclosure was examined in C57BL / 6J mice. In certain embodiments, mice were intravenously (1 µg mRNA / mouse) or intramuscularly (2 µg mRNA / mouse) administered LNPs of the disclosure, or controls (e.g., SM-102), comprising luciferase mRNA and the distribution thereof to the liver compared to the full body was evaluated (FIGs.3A-3C). In one aspect, these experiments demonstrated that the LNPs of the disclosure perform in a manner greater than or equal to SM-102, while minimizing delivery to the liver. Additionally, distribution of LNPs of the disclosure, as compared to controls (e.g., SM-102), to the inguinal and iliac lymph nodes was examined (FIGs.4A-4C). In another, aspect, these experiments demonstrated that the LNPs of the disclosure perform better than LNPs comprising non- aromatic ionizable lipids in transfecting lymph nodes and / or perform comparably to SM-102 - 117 - 53224792.3 Attorney Docket No.0466483-7483WO1 in this regard (see also FIGs.5A-5C). Additionally, the LNPs of the disclosure comprising aromatic ionizable lipids appear to be less liver-tropic than SM-102, yet exhibit similar lymphatic tropism (FIGs.6A-6C). Example 5: Therapeutic application – delivery of SARS-CoV-2 RBD mRNA The disclosure further describes an exemplary therapeutic application of the LNPs of the disclosure. In one non-limiting embodiment, a dosing scheme for a COVID vaccination was designed utilizing C57BL / 6J mice (FIG.7). In certain embodiments, mice are primed on day 0 (D0), administered a second dose (boost) on day 21 (D21) and sacrificed on day 40 (D40). Additionally, samples were collected periodically throughout the dosing regime. Mice were administered either 1 µg or 5 µg of COVID receptor-binding domain (RBD) mRNA, wherein the mRNA was encapsulated in an LNP (e.g., SM-102, C8mPhE-383, or C6pPhE- 383). Three (3) weeks post-boost, COVID RBD IgG antibody titers in mice administered certain LNPs of the disclosure were measured and found to be comparable to those observed with administration of the comparator or control LNP (i.e., SM-102 LNP) (FIG.8). Further, substantial toxicity was not observed with administration of the LNPs of the disclosure encapsulating RBD mRNA. Thus, in one aspect, the experiments described herein demonstrate the utility of the LNPs described herein for mRNA delivery, for uses including, but not limited to, vaccination (e.g., COVID vaccination). Materials and Methods (Examples 6-12) Materials For chemical synthesis, Dimethyl dipimidate (Sigma-Aldrich), Tris(2- cyanoethyl)amine (TCI), butane-1,4-diamine (Sigma-Aldrich), acrylonitrile (Sigma-Aldrich) hydrogen chloride (Sigma-Aldrich), Sodium bicarbonate (Sigma-Aldrich), NaCl (Sigma- Aldrich), Diethylenetriamine (Sigma-Aldrich), 1,3-diaminopropane (Sigma-Aldrich), 2,2'- Diamino-N-methyldiethylamine (TCI), Triethylentetramine (Sigma-Aldrich), 2-(2- Aminoethylamino)ethanol (Sigma-Aldrich), 3,3'-Diamino-N-methyldipropylamine (Sigma- Aldrich), Tetraethylene pentamine (Sigma-Aldrich), 2,2'-Disulfanediyldiethanamine (AmBeed), N,N-dimethyldipropylenetriamine (Sigma-Aldrich), Pentaethylenehexamine (Sigma-Aldrich), 1,2-Bis(2-aminoethoxy)ethane (TCI), N-Methyl-1,3-diaminopropane (Sigma-Aldrich), Aminoethylpiperazine (Sigma-Aldrich), 1,4-bis(3-aminopropyl)piperazine (Sigma-Aldrich), N1-(2-(4-(2-Aminoethyl)piperazin-1-yl)ethyl)ethane-1,2-diamine (AmBeed), p-Xylylenediamine (Sigma-Aldrich), 3-(4-{2-[(3-amino-2- - 118 - 53224792.3 Attorney Docket No.0466483-7483WO1 ethoxypropyl)amino]ethyl}piperazin-1-yl)-2-ethoxypropan-1-amine (Enamine), 2-{2-[4-(2- {[2-(2-aminoethoxy)ethyl]amino}ethyl)piperazin-1-yl]ethoxy}ethan-1-amine (Ena-mine), 1- N-{2-[4-(4-aminocyclohexyl)piperazin-1-yl]ethyl}cyclohexane-1,4-diamine (Enam-ine), {2- [2-(2-aminoethoxy)ethoxy]ethyl}[2-(4-{2-[2-(2-aminoethoxy)ethoxy]ethyl}-piperaz-in1- yl)ethyl]amine (Enamine), Triethylamine (TEA, Sigma-Aldrich), 1,2-Epoxydodecane (Sigma-Aldrich), and solvents for synthesis were purchased from commercial sources and were used as received. For LNP formulation, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE, Avanti Research), cholesterol (Sigma-Aldrich), and 1,2-dimyristoyl-sn-glycero-3- phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (C14-PEG2000, Avanti Research) were purchased from commercial sources. For BMDCs differentiation, Iscove’s modified Dulbecco’s medium (Gibco), fetal bovine serum (Gibco), and Penicillin- Streptomycin (Gibco), mouse IL-4 (GenScript), mouse GM-CSF (GenScript), and β- mercaptoethanol (Sigma-Aldrich) were purchased from commercial sources. For in vitro studies, Luicferase Reporter 1000 Assay System (Promega), CellTiter-Glo assay (Promega), Hoechst 33342 (ThermoFisher Scientific), LysoTracker™ Green DND-26 (Invitrogen) were purchased from commercial sources. B16-OVA melanoma cell lines and hACE-HEK293 cells were purchased from Sigma-Aldrich and InvivoGen, respectively. For western blot, RIPA lysis buffer (ThermoFisher Scientific), Halt™ Phosphatase Inhibitor Cocktail (ThermoFisher Scientific), Halt™ Protease Inhibitor Cocktail (ThermoFisher Scientific), anti-S6K antibody (Cell Signaling Technology, #9202), anti-phospho-S6K antibody (Cell Signaling Technology, #9205), anti-Akt antibody (Cell Signaling Technology, #4691), anti- phospho-Akt antibody (Cell Signaling Technology, #4060) and anti-b-actin (Cell Signaling Technology, #4970), IRDye 800CW Goat anti-Rabbit IgG (LI-COR) were purchased from commercial sources. For RNA extraction, PureLink RNA Mini Kit was purchased from Invitrogen. For flow cytometry, anti-mouse CD16 / 32 antibody (BD Bioscience), LIVE / DEAD™ Fixable Aqua Dead Cell Stain Kit (ThermoFisher Scientific), Cell Activation Cocktail (BioLegend), Cyto-Fast™ Fix / Perm Buffer Set (BioLegend), APC-conjugated H- 2Kb / OVA (SIINFEKL) MHC Tetramer (MBL Life Science), biotinylated SARS-CoV-2 Spike RBD protein (Sino Biological), and SARS-CoV-2 RBD peptide pools (#PM-WCPV-S-RBD- 1, JPT Peptide Technologies) were purchased from commercial sources. PE-CF594- conugated anti-mouse CD3 antibody, Brilliant Violet (BV) 421-conugated anti-mouse CD8 antibody, PE-conjugated anti-mouse IFN-g antibody, APC-conugated anti-mouse TNF-a antibody, BV650-conjugated anti-mouse CD80 antibody, BV711-conjugated anti-mouse IgD antibody, BV785-conjugated anti-mouse B220 antibody, Alexa Fluor (AF) 700-conjugated - 119 - 53224792.3 Attorney Docket No.0466483-7483WO1 anti-mouse CD38 antibody, PE-conjugated anti-mouse PD-L2 antibody, PE-CF694- conjugated anti-mouse IgM antibody, PE-Cy5.5-conjugated anti-mouse CD19 antibody, PE- Cy7-conjugated anti-mouse GL7 antibody, BV421-conjugated Streptavidin, AF647- conjugated Streptavidin, BV785-conjugated anti-mouse CD3 antibody, FITC-conjugated anti-mouse CD4 antibody, PE-conjugated anti-mouse CXCR5 antibody, PE-Cy5-conjugated anti-mouse CD44 antibody, BV421-conjugated anti-mouse CD62L antibody, FITC- conjugated anti-mouse CD3 antibody, and PE-conjugated anti-mouse CD8 antibody were purchased from BioLegend. For tissue immunofluorescence staining, AF 488-cojugated anti- mouse GL7 antibody, AF 594-cojugated anti-mouse CD21 antibody, AF 647-cojugated anti- mouse B220 antibody, AF 488-cojugated anti-mouse IFN-g antibody, and AF 647-cojugated anti-mouse CD8a antibody were purchased from BioLegend. For ELISA, RBD protein (Sino Biological), OVA protein (InvivoGen), and High Bind Stripwell 96-well clear polystyrene microplates (Corning) were purchased from commercial sources. For ELISpot, mouse IFN-g ELISpot Kit (R&D systems), OVA257-264peptide (InvivoGen), MultiScreenHTS IP Filter Plate (Sigma-Aldrich) biotinylated anti-mouse IgG antibody (Southern Biotech), streptavidin- alkaline phosphatase (ThermoFisher Scientific), 5-bromo-4-chloro-3-indolyl-phosphate / nitro blue tetrazolium chloride solution (Sigma-Aldrich), and sodium phosphate monobasic (Sigma-Aldrich) were purchased from commercial sources. mRNA synthesis Codon-optimized firefly luciferase, green fluorescent protein, ovalbumin, or SARS- CoV-2 spike protein sequences were inserted into a template plasmid with optimized 3’ and 5’ UTR and 101 poly A tail. mRNAs were synthesized through in vitro transcription of the plasmids in the presence of 1-methyl pseudouridine modified nucleosides and CleanCap (TriLink BioTechnology) capping. After cellulose-based purification, mRNAs were ethanol- precipitated, washed, re-suspended in nuclease-free water and subjected to quality control (gel electrophoresis, standard J2 dot blot, and endotoxin content). All mRNAs were stored at -20 °C until use. LNP formulation The corresponding ionizable lipid, DOPE, cholesterol and C14-PEG2000 were dissolved in ethanol at a molar ratio of 57.1:10.2:31.1:1.7, respectively, to form the organic phase. For the ALC-0315 LNP, the organic phase was prepared in ethanol at a molar ratio of 50:10:38.5:1.5, respectively. An aqueous phase was prepared consisting of the corresponding - 120 - 53224792.3 Attorney Docket No.0466483-7483WO1 mRNA in a 10 mM citrate buffer at pH 3. LNPs were formulated by mixing the organic phase and aqueous phase at a volume ratio of 1:3, either by pipette mixing for the library screening or microfluidic mixing for the other experiments. LNPs were dialyzed against 1× PBS in a 20-kDa molecular-weight-cutoff cassette for 2 h. Molecular structure and simulation 3D molecular structures of ionizable lipids were predicted and visualized using Gaussian 16 software. For 3D modeling, the molecular geometry was optimized through the Universal Force Field. The angle between the lipid-attached amines and the distance between amines in the optimized structure were calculated. The molecular electrostatic potential was analyzed using Jmol software. For the simulation of lipids interactions, each structure of the ionizable lipid, DOPE and C14-PEG2000 was optimized as mentioned above, and interaction simulation were performed using Gaussian 16. Single-point energy calculation was carried out. LNP characterization LNPs were characterized with respect to particle morphology, size, zeta potential, pKa, and mRNA encapsulation efficiency. The morphology of LNPs was observed by cryo-EM (Titan Krios, Thermo Fisher). The hydrodynamic size, PDI, and zeta potential of LNPs were measured using a Zetasizer Nano ZS90 (Malvern Instruments). The pKa of LNPs was determined using a 6-(p-toluidinyl)naphthalene-2-sulfonic acid (TNS) assay. The mRNA encapsulation efficiency was determined by the Quant-iT RiboGreen RNA assay (Invitrogen). Isolation of bone marrow-derived dendritic cells (BMDCs) Bone marrow cells were isolated from the femurs and tibias collected from 6-week-old C57BL / 6 mice. The isolated cells were incubated in Iscove’s modified Dulbecco’s medium supplemented with 10% fetal bovine serum, 100 mg / mL streptomycin, 100 units / mL penicillin, 20 ng / mL recombinant mouse granulocyte-macrophage colony-stimulating factor, 20 ng / mL recombinant mouse interleukin-4, and 50 μM β-mercaptoethanol for dendritic cell differentiation. BMDCs were ready for use on day 7. In vitro mRNA transfection For LNP library screening, BMDCs were seed into 96-well plates at a density of 2×105cells per well overnight. The cells were treated with mLuc loaded LNPs (125 ng of mRNA - 121 - 53224792.3 Attorney Docket No.0466483-7483WO1 per well) and incubated for 6 h. The luciferase mRNA expression was evaluated using the Luicferase Reporter 1000 Assay System according to the manufacturer’s instructions, and the luminescence intensity was quantified using an Infinite 200 Pro plate reader (Tecan). For imaging of endosomal escape, BMDCs were seed into 8-well confocal imaging plates at a density of 4×105cells per well overnight. The cells were treated with Cy5-labeld mRNA-loaded LNPs (250 ng of mRNA per well) and incubated for 6 h. Cells were stained with Lysotracker Green DND-26 (50 nM) for 30 min and Hoechst 33342 (10 µg / ml) for 5 min. The cells were imaged using a confocal laser scanning microscope (LSM 710, Zeiss). For imaging of mRNA expression, BMDCs were seed into 8-well confocal imaging plates at a density of 4×105cells per well overnight. The cells were treated with mGFP- loaded LNPs (250 ng of mRNA per well) and incubated for 24 h. Cells were stained with Hoechst 33342 (10 µg / ml) for 5 min and the mRNA expression was observed by using a confocal laser scanning microscope. For antigen presentation, BMDCs were seed into 24-well plates at a density of 5×105cells per well overnight. The cells were treated with mOVA loaded LNPs (500 ng of mRNA per well) and incubated for 24 h. Cells were collected and blocked with anti-mouse CD16 / 32 antibody, followed by staining with FITC-conjugated anti-mouse CD11c antibody and APC- conjugated anti-mouse H-2Kbbound to SIINFEKL Antibody for 30 min. The cells were analyzed by flow cytometry using an LSR II Flow Cytometer (BD Biosciences). For in vitro cell viability assessment, BMDCs were seed into 96-well plates at a density of 2×105cells per well overnight. The cells were treated with mLuc loaded LNPs (125 ng of mRNA per well) and incubated for 24 h. The cell viability was measured using a CellTiter-Glo cell viability assay according to the manufacturer’s instructions, and the luminescence intensity was quantified using an Infinite 200 Pro plate reader. Seahorse assays BMDCs treated with mLuc-loaded LNPs for 24 h were seeded into Seahorse XFe96 / XF Pro Cell Culture Microplates at a density of 2×105cells per well. The extracellular acidification rate and oxygen consumption rate of the cells were measured by using Seahorse XF Glycolysis Stress Test kit, Cell Mito Stress Test kit, and Real-Time ATP Rate Assay kit according to the manufacturer’s instructions. For the Glycolysis Stress Test, 10 mM of glucose, 2.5 mM of oligomycin, and 50 mM of 2-DG were injected. For the Cell Mito Stress Test, 1 mM of FCCP and 0.5 mM of rotenone / antimycin A were injected. For the Real-Time ATP Rate Assay, 2.5 mM of oligomycin and 0.5 mM of rotenone / antimycin A were injected - 122 - 53224792.3 Attorney Docket No.0466483-7483WO1 and analyzed using XF96 Extracellular Flux Analyzer (Agilent). Western blot BMDCs treated with mLuc-loaded LNPs or LPS (1 µg / ml) for 30 min were collected and lysed with RIPA lysis buffer containing a protease inhibitor cocktail and a phosphatase inhibitor cocktail on ice for 30 min. The cell lysates were centrifuged at 15,000 g for 30 min, and the supernatants were collected. The amount of protein was quantified using the DC Protein Assay. The lysates were separated by 4-12% Bis-Tris gels and transferred to a nitrocellulose membrane. For primary antibodies, anti-S6K, anti-phospho-S6K, anti-Akt, anti-phospho-Akt and anti-b-actin were used. IRDye 800CW goat anti-rabbit IgG was used as the secondary antibody. Protein detection was measured using the Odyssey M Imager (LI- COR). mRNA-Seq BMDCs were treated with mLuc-loaded LNPs for 6 h, and total RNA was extracted using the PureLink RNA Mini Kit. RNA library preparation, sequencing, and data analysis were conducted at Azenta Life Sciences (South Plainfield, NJ, USA). RNA sequencing libraries were prepared using the NEBNext Ultra II RNA Library Prep Kit for Illumina using manufacturer’s instructions (NEB, Ipswich, MA, USA). The sequencing libraries were multiplexed and clustered onto a flowcell on the Illumina NovaSeq instrument according to manufacturer’s instructions. The samples were sequenced using a 2x150bp Paired End (PE) configuration. Using DESeq2, a comparison of gene expression between the groups of samples was performed. The Wald test was used to generate p-values and Log2 fold changes. Genes with adjusted p-values < 0.05 and absolute log2 fold changes > 1 were called differentially expressed genes for each comparison. Gene set enrichment analysis was performed using GSEA v4.3.3. KEGG pathway enrichment analysis was performed using ShinyGO v0.80. In vivo study C57BL / 6 female mice (6 weeks old, 18–20 g) were purchased from Jackson Laboratory. In vivo immunization Immunization was conducted following a prime-boost strategy at a 3-week interval for - 123 - 53224792.3 Attorney Docket No.0466483-7483WO1 the SARS-CoV-2 model and a 5-day interval for the OVA cancer model. For each injection, mice were intramuscularly injected with mRBD or mOVA-loaded LNPs at a dose of 2 mg of mRNA. For serum analysis, blood was collected into serum-separating tubes via the retro- orbital route and centrifuged at 10,000 × g for 5 min. The serum was stored at −20 °C until use. For imaging of mRNA expression, mLuc-loaded LNPs were intramuscularly injected at a dose of 2 mg of mRNA. At 6, 24, 48 and 72 h time points, mice were intraperitoneally injected with D-luciferin potassium salt at a dose of 150 mg / kg, and bioluminescence at the injection site was measured using an IVIS imaging system (PerkinElmer). Humoral immune responses Vaccine-induced humoral immune responses were evaluated with respect to antigen- specific antibody titers, antigen-specific B cell induction, and pseudovirus neutralization assay. For antibody titers determination by ELISA, High Bind Stripwell Corning 96-well clear polystyrene microplates were coated with SARS-CoV-2 RBD or OVA protein (1 µg / ml). After overnight incubation, plates were washed with 0.05 % Tween 20 in PBS (PBST) and blocked for 2 h at r.t. using 2% IgG-depleted bovine serum albumin in PBS. After washing with PBST, diluted sera in blocking buffer were added to each well and incubated for 2 h at r.t. Plates were washed again with PBST and incubated with horseradish peroxidase- conjugated anti-mouse IgG in blocking buffer for 2 h at r.t. Plates were washed thoroughly with PBST and 3,3′,5,5′-tetramethylbenzidine substrate was added. The reaction was stopped after 10 min incubation with 2 N sulfuric acid, and the absorbance was detected at 450 nm using an Infinite 200 Pro plate reader. For the analysis of antigen-specific B cells, splenocytes were isolated from the spleen of immunized mice by filtering through a 70 µm cell strainer and performing red blood cell lysis. For antibody staining, splenocytes were blocked with anti-mouse CD16 / 32 antibody for 20 min and then stained with BV711-conjugated anti-mouse IgD antibody, BV785- conjugated anti-mouse B220 antibody, PE-CF694-conjugated anti-mouse IgM antibody, PE- Cy5.5-conjugated anti-mouse CD19 antibody, biotinylated RBD, BV421-conjugated Streptavidin, and AF647-conjugated Streptavidin for 30 min. The cells were analyzed by flow cytometry using a LSR II Flow Cytometer. For the pseudovirus neutralization assay, hACE2-HEK cells were seeded into 96-well plates at a density of 1×105cells per well overnight. Diluted serum samples were incubated with SARS-CoV-2 pseudovirus for 1 h at 37 °C. Afterwards, the antibody-pseudovirus - 124 - 53224792.3 Attorney Docket No.0466483-7483WO1 mixture was treated to hACE2-HEK cells and incubated for 24 h. Infection of pseudovirus was measured by luciferase expression using a Luicferase Reporter 1000 Assay System, and the 50% neutralization titer was determined as the highest serum dilution at which neutralization was reduced by at least 50% relative to control cells that were infected with pseudovirus in the absence of serum. Cellular immune responses Vaccine-induced cellular immune responses were evaluated with respect to antigen- specific T cells in the spleen. For the ELISpot assay, splenocytes were isolated from spleen of immunized mice by filtering through a 70 µm cell strainer and performing red blood cell lysis. The splenocytes were seeded onto IFN-g ELISPot plates (R&D systems) at a density of 1 × 106cells per well and stimulated with SARS-CoV-2 RBD peptide pools (2 µg / ml) or the OVA peptide epitope (OVA(257–264) , 10 µg / ml) in complete RPMI at 37 °C for 24 h. Spots of IFN-g-secreting cells were developed according to the manufacturer’s instructions and counted using a CTL ImmunoSpot S3 analyzer. For OVA-specific CD8+T cells, splenocytes were blocked using anti-mouse CD16 / 32 antibody for 20 min, and stained with FITC-conjugated anti-mouse CD3 antibody, PE- conjugated anti-mouse CD8 antibody, and APC-conjugated H-2Kb OVA(SIINFEKL) (SEQ ID NO:1) tetramer for 30 min. The cells were then analyzed by flow cytometry using an LSR II Flow Cytometer. For intracellular cytokine analysis, splenocytes were seeded into 24-well plates and stimulated with SARS-CoV-2 RBD peptide pools (2 µg / ml). A cell activation cocktail containing phorbol-12-myristate 13-acetate (81 nM), ionomycin (1.33 mM), and Brefeldin A (5 µg / ml) was added 1 h after stimulation. After a total of 6 h of stimulation, the cells were blocked using anti-mouse CD16 / 32 antibody for 20 min, followed by staining with PE- CF594-conugated anti-mouse CD3 antibody and BV421-conugated anti-mouse CD8 antibody for 30 min. For intracellular markers, cells were fixed and permeabilized using Cyto-Fast™ Fix / Perm Buffer Set and stained with PE-conjugated anti-mouse IFN-g antibody and APC- conjugated anti-mouse TNF-a antibody for 30 min. The cells were analyzed by flow cytometry using an LSR II Flow Cytometer. Memory immune response Vaccine-induced memory immune responses were evaluated with respect to memory cell induction, germinal center (GC) formation, and plasma cells in the bone marrow. For GC - 125 - 53224792.3 Attorney Docket No.0466483-7483WO1 B cells, and memory T cells, spleens were collected and processed as single cells for flow cytometry by filtering through a 70 µm cell strainer and performing red blood cell lysis. The antigen-specific B cells in the spleen were further analyzed by staining with BV650- conjugated anti-mouse CD80 antibody, AF700-conjugated anti-mouse CD38 antibody, PE- conjugated anti-mouse PD-L2 antibody, and PE-Cy7-conjugated anti-mouse GL7 antibody staining. The formation of GCs in the lymph nodes was evaluated using tissue immunofluorescence. Collected inguinal lymph nodes were fixed in 4% paraformaldehyde for 4 h and transferred to 30% sucrose for cryoprotection overnight. Afterwards, the tissues were embedded in the OCT compound, frozen and sectioned to a thickness of 4 mm. For immunostaining, the samples were blocked with 2.5% horse serum for 1 h, followed by staining with 1:200 dilutions of the AF488-conjugated anti-mouse GL7 antibody, AF594- conjugated anti-mouse CD21 and AF647-conjugated anti-mouse B220. After overnight incubation, slides were washed with PBST and mounted using VECTASHIELD Plus Antifade Mounting Medium with DAPI (Vector Laboratories). The samples were imaged by using Stellaris 5 confocal microscopy (Leica). Plasma cell induction in the bone marrow was measured by ELISpot assay. Bone marrows were isolated from the femurs and tibias of immunized mice and processed into a single-cell suspension by filtering through a 70 µm cell strainer and performing red blood cell lysis. Cells were seeded onto RBD-coated (10 µg / ml) MultiScreenHTS IP filter plates at a density of 1 × 106cells per well in complete RPMI. After 24 h of incubation at 37 °C, plates were washed with PBST, and biotinylated anti-mouse IgG antibody (3 µg / ml in 2% BSA / PBS) was added to each well and incubated at r.t. for 1 h. The plates were then washed with PBST, and streptavidin-alkaline phosphatase (1:20,000 dilution in 2% BSA / PBS) was added to each well and incubated at r.t. for 30 min. After thorough washing with PBST, the spots were developed by adding 50 µl of 5-bromo-4-chloro-3-indolyl-phosphate / nitro blue tetrazolium chloride solution to each well. The reaction was quenched by adding 100 µl of 1 M sodium phosphate monobasic to each well, and the plates were washed with deionized H2O and dried overnight. Spots of antibody-secreting cells were counted using a CTL ImmunoSpot S3 analyzer. In vivo efficacy of cancer vaccine model The therapeutic efficacy of the B16-OVA cancer vaccine model was evaluated with respect to tumor volume, survival rate, and tumor-infiltrating lymphocyte analysis. For the - 126 - 53224792.3 Attorney Docket No.0466483-7483WO1 establishment of the tumor model, C57BL / 6 mice were subcutaneously inoculated with B16- OVA cells (1 × 105cells for preventive cancer vaccine model and 5 × 105cells for therapeutic cancer vaccine model). Tumor volume was calculated as A × B2× 0.5, where A and B are the lengths of the largest and smallest dimensions, respectively. Tumor growth and mice survival were monitored for up to 44 days. For tumor-infiltrating lymphocyte analysis, tumors collected on day 15 from the therapeutic cancer vaccine model were processed into single-cell suspensions. Cells were treated with a cell activation cocktail containing phorbol-12-myristate 13-acetate (81 nM), ionomycin (1.33 mM), and Brefeldin A (5 µg / ml) for 6 h, followed by blocking with anti- mouse CD16 / 32 antibody for 20 min and staining with APC-conjugated anti-mouse CD3 antibody and BV421-conjugated anti-mouse CD8 antibody. For IFN-g staining, cells were fixed and permeabilized using Cyto-Fast™ Fix / Perm Buffer Set and stained with PE- conjugated anti-mouse IFN-g antibody for 30 min. The cells were analyzed by flow cytometry using a LSR II Flow Cytometer. For imaging of tumor tissue, collected tumors were processed and measured according to the lymph node sample preparation method mentioned above. Samples were stained with 1:200 dilutions of the AF488-conjugated anti- mouse IFN-g antibody and AF647-conjugated anti-mouse CD8a. Statistics All statistical analyses were performed using GraphPad Prism 8 software. All data are presented as mean ± s.d. To assess statistical differences between two groups, the Student's t- test was applied. To assess statistical differences among multiple groups, a one-way ANOVA followed by Tukey’s test was applied. In all statistical tests, p < 0.05 was considered statistically significant. Example 6: Compound Synthesis Synthesis of imidoester linker Dimethyl adipimidate was linker. The compound was characterized by1H NMR using a Bruker 400-MHz NMR spectrometer.1H NMR (400 MHz, Deuterium Oxide) δ 3.65 (d, J = 2.3 Hz, 6H), 2.38 (dt, J = 8.5, 3.9 Hz, 4H), 1.59 (dp, J = 7.0, 4.2, 3.5 Hz, 4H). - 127 - 53224792.3 Attorney Docket No.0466483-7483WO1 g, 20 mmol) (50 mL), and the mixture was cooled to 4 °C in an ice bath. Anhydrous HCl gas (2.62 g, 72 mmol) was slowly added to the solution for 4 h. The reaction was then kept at 4 °C for 48 h. The volatiles were removed under vacuum, and the mixture was dispersed in dichloromethane (50 ml) and washed with saturated sodium bicarbonate (2 × 50 ml) and brine (2 × 50 ml). The organic layer was collected, dried over Na2SO4 and concentrated in vacuo. The product was isolated as a white powder with a 48.5 % yield. The compound was characterized by1H NMR using a Bruker 400-MHz NMR spectrometer.1H NMR (400 MHz, Chloroform-d) δ 3.66 (s, 9H), 2.76 (t, J = 7.1 Hz, 6H), 2.44 (t, J = 7.1 Hz, 6H). diylbis(azanetriyl))tetra-propanenitrile (a). For the synthesis of a, butane-1,4-diamine (2.20 g, 25 mmol) was dissolved in 25 ml of water, and acrylonitrile (10.61 g, 200 mmol) was added dropwise to the solution. The mixture was heated at 80 °C for 1 h, and the reaction was kept at 40 °C for 12 h. The product was collected by filtration, and excess of acrylonitrile was removed under vacuum. The product was isolated as a white powder with a 89.8 % yield.1H NMR (400 MHz, Chloroform-d) δ 2.88 (t, J = 6.6 Hz, 8H), 2.67 – 2.56 (m, 4H), 2.50 (t, J = 6.6 Hz, 8H), 1.62 – 1.53 (m, 4H). For the 4-arm imidoester linker synthesis (b), a (3.00 g, 10 mmol) and methanol (1.53 g, 48 mmol) were dissolved in anhydrous dichloromethane (50 mL) and the mixture was cooled to 4 °C in an ice bath. Anhydrous HCl gas (1.75 g, 48 mmol) was slowly added to the solution for 4 h. The reaction was then kept at 4 °C for 48 h. The volatiles were removed under vacuum and the mixture was dispersed in dichloromethane (50 ml) and washed with saturated sodium bicarbonate (2 × 50 ml) and brine (2 × 50 ml). The organic layer was - 128 - 53224792.3 Attorney Docket No.0466483-7483WO1 collected, dried over Na2SO4 and concentrated in vacuo. The product was isolated as a white powder with a 32.7 % yield. The compound was characterized by1H NMR using a Bruker 400-MHz NMR spectrometer.1H NMR (400 MHz, Deuterium Oxide) δ 3.69 (s, 12H), 3.48 (t, J = 6.7 Hz, 8H), 3.25 (d, J = 7.5 Hz, 4H), 2.90 (t, J = 6.7 Hz, 8H), 1.84 – 1.76 (m, 4H). Synthesis of ionizable lipids The crosslinked ionizable lipids were prepared through a one-pot, two-step synthesis. For crosslinking the amine compounds, the linkers and amines were mixed at a molar ratio of 1:1.1 between the imidoester functional group and primary amine group. (0.25 mmol, 5 equiv.) were dissolved in ethanol and 3,3'-(piperazine-1,4-diyl)bis(propan-1- amine) (0.11 mmol, 2.2 equiv.) was added and stirred for 2 h at r.t. Next, the crosslinked amine cores were reacted with excess epoxide lipid tails to saturate the amines. Taking C12- 2aN as an example 1,2-Epoxydodecane (0.33 mol, 6.6 equiv.) was added to the mixture and heated at 80 °C for 48 h. The volatiles were removed under vacuum and the crude product was used for the initial library screening. C12-N and C12-2aN were further purified using the CombiFlash NextGen 300+ chromatography system (Teledyne ISCO) with gradient elution from dichloromethane to 80:20 dicholoromethane / methanol. The products were isolated as a clear oil with 67.2 % and 53.7 % yield, respectively. For lipid characterization, LC-MS was performed using a Waters Acquity LCMS system equipped with UV-Vis and MS detectors (calculated MS of C12-N, 937.62; found [M + 2H]2+, 469.76, calculated MS of C12-2aN, 1246.10; found [M + H]+, 1247.52). Example 7: Imidoester crosslinkers enables the high-throughput synthesis of ionizable lipids for mRNA delivery to BMDCs - 129 - 53224792.3 Attorney Docket No.0466483-7483WO1 Using rapid and straightforward conjugation chemistry, 2-arm (2a), 3-arm (3a), or 4- arm (4a) imidoester crosslinkers and amine groups in the amine compounds were reacted to create various multi-amine core structures. C12 lipid tails were then attached to the remaining amine groups via an epoxide ring-opening reaction (FIG.13A). This one-pot two-step reaction was used to generate a library of novel ionizable lipids in a high throughput manner. To prepare the library, a range of amine core structures were used, including 12 linear- structured amines and 8 cyclic-structured amines (FIG.13B). To synthesize control ionizable lipids, the corresponding amines were directly conjugated with C12 lipid tails under the same reaction conditions, but without the crosslinkers. The ionizable lipids were then combined with DOPE, cholesterol, and C14-PEG and formulated into LNPs containing luciferase mRNA (mLuc) for screening in bone marrow-derived dendritic cells (BMDCs) (FIG.13C). The reporter mRNA-based screening results revealed that the crosslinked ionizable lipids improved mRNA transfection in BMDCs (FIG.13D and FIG.14B). Notably, the groups treated with 2a linker-based LNPs showed a significant increase in mRNA expression across the amine compounds compared to the control LNP-treated groups. With the 2a linker, LNPs with 16 out of the 20 amine cores showed improved luminescence intensity (FIG. 14C). Notably, the 2a-based LNP-treated groups based on amine J and amine N, the respective top-performing amine cores for linear and circular amine structures, facilitated 7.31- and 21.7-fold higher mRNA expression level compared to the groups treated with corresponding control LNPs (FIG.14D). On the other hand, there were four amine cores (E, I, L and M) which demonstrated decreased luminescence intensity in the 2a linker-based LNP-treated groups compared to the corresponding control LNP-treated groups (FIGs.14E-14F). These amine cores share the common characteristic of not having any primary amine groups remaining after the conjugation reaction with imidoester. The attachment of only one lipid tail to the remaining secondary amine group of each core was found to be insufficient, indicating that the conjugation of multiple lipid tails to the primary amine groups of amine cores is an important factor for effective mRNA transfection. Through the in vtiro screening, C12-2aN, an ionizable lipid with crosslinked amine N and 2a linker, was selected as the top-performer due to its highest mRNA transfection efficiency. Example 8: Incorporation of crosslinkers affects the structural and electrostatic properties of ionizable lipids for mRNA delivery Focusing on amine N, the structure of ionizable lipids according to linker type was - 130 - 53224792.3 Attorney Docket No.0466483-7483WO1 examined to uncover what mechanistically enables the high transfection efficiency of C12- 2aN. Analysis of 3D structures of the ionizable lipid revealed that the incorporation of crosslinkers affects the arrangement of amine cores and has a significant impact on the conformation of lipids (FIGs.15A-15B). While C12-N has a chair conformation with an angle of 151.34° between the lipid attached amines, showing that ionizable amines are arranged horizontally, C12-2aN has a bent structure with a 48.12° angle, indicating that ionizable amines are biased to one side and protrude. C12-3aN and C12-4aN have out-of- plane bending structures with the angles of 120.74 ° and 111.06 °, respectively, showing relatively moderate bending. The linker not only affected the structural conformation of the ionizable lipid but also influenced the electrostatic properties. Since the crosslinkers introduce basic amidine functional groups, they exhibit relatively lower electrostatic potential compared to other parts of the ionizable lipid. In the molecular electrostatic potential map of C12-2aN, the bent and protruding core region showed relatively low electrostatic potential, leading to an asymmetric distribution compared to the evenly distributed electrostatic potential of C12-N. These ionizable lipid properties also affected the lipid arrangement in LNPs. Structural simulation of the complete LNPs revealed that the amine core of C12-2aN protrudes on the surface while interacting with other excipient lipids, which may strengthen the interaction between LNPs and negatively charged cell membranes (FIG.15C and FIGs. 16A-16B). As a result of the changes in structural and electrostatic properties of the ionizable lipid and its arrangement on the LNP, LNPs with different crosslinkers showed varying levels of mRNA endosomal escaped to the cytosol. Consistent with the in vitro screening results, C12-2aN showed the most efficient endosomal escape as indicated by lesser colocalization of lysotracker and Cy5-mRNA, resulting in the highest mRNA expression in BMDCs (FIG.15D and FIGs.16C-16D). Example 9: C12-2aN determined characteristics of LNP for mRNA vaccine Further investigation was conducted to determine if C12-2aNP, the lead LNP formulation based on the C12-2aN, is suitable for vaccine applications. As shown in the chemical structure, the 2a linker created two amidine groups between the ionizable piperazines in C12-2aN (FIG.17A and FIG.19A). The insertion of amidines resulted in a positive surface charge of the nanoparticle. While the C12-N-based LNP (C12-NP) showed a negative zeta potential of -11.64 ± 0.47 mV, C12-2aNP exhibited a positive zeta potential of +7.50 ± 0.57 mV (FIG.17B). The incorporation of the 2a linker also affected the pKa of the - 131 - 53224792.3 Attorney Docket No.0466483-7483WO1 lipid. A TNS assay revealed that C12-2aN has a pKa of 7.12, while C12-N has a pKa of 6.19, indicating that the lipids are ionizable at different physiological pH levels (FIG.17C). On the other hand, both C12-N and C12-2aN exhibited similar characteristics in terms of particle formation. The spherical morphology filled with lipid layers at the core for both C12-NP and C12-2aNP was observed by cryo-TEM (FIG.17D). The mean particle sizes of C12-NP and C12-2aNP were 110.2 ± 2.3 nm and 92.0 ± 2.5 nm, respectively (FIG.17E). Polydispersity index (PDI) of C12-N and C12-2aNP were 0.155 ± 0.032 mV and 0.204 ± 0.038 mV, indicating that both particle size distributions are monodisperse (FIG.17F). The mRNA encapsulation efficiencies in C12-NP and C12-2aNP were 83.85 ± 1.28 % and 90.11 ± 1.13 %, respectively, indicating that mRNA is efficiently loaded during formulation of both LNPs (FIG.17G). No significant difference in in vitro toxicity was observed between C12- NP and C12-2aNP in BMDCs, where the cell viabilities were 86.66 ± 1.40 % and 88.17 ± 2.66 %, respectively (FIG.17H). After characterization, the potential for C12-2aNP to serve as an effective mRNA vaccine was evaluated in vivo by intramuscular injection. mLuc-loaded LNPs were injected into mouse thigh muscles at an mRNA dose of 0.1 mg / kg, and whole-body bioluminescence imaging was conducted at various timepoints. At 6 h, bioluminescence intensity in C12-2aNP treated mice was 3.24-fold higher than that of C12-NP (FIG.17I). Additionally, monitoring the bioluminescence over 72 h revealed that the area under curve (AUC) of mRNA expression in C12-2aNP-treated group was 3.15-fold higher than that in the C12-NP-treated group (FIGs.17J-17K) Example 10: C12-2aNP induces a metabolic switch to glycolysis in BMDCs Next, the ability of C12-2aNP to stimulate glycolysis in dendritic cells was evaluated, a switch which could lead to more potent vaccination. When BMDCs were treated with mLuc-loaded C12-2aNP, a significant decrease in the pH of the cell media was observed. After 24 h of LNP incubation, the pH of the media in the C12-2aNP-treated group dropped from 7.56 ± 0.00 to 6.52 ± 0.04, while in the untreated and C12-NP-treated groups, the pH only decreased to 7.23 ± 0.04 and 7.16 ± 0.02, respectively (FIGs.18A-18B). As the acidification of the medium is indicative of proton and lactate secretion resulting from an elevated cellular metabolic rate, the activity of central metabolic regulator mammalian target of rapamycin (mTOR) was further examined to investigate the underlying mechanism. Since mTOR is a protein kinase that forms two distinct protein complexes, mTORC1 and mTORC2, which are involved in two major signaling pathways for metabolic regulation, the - 132 - 53224792.3 Attorney Docket No.0466483-7483WO1 phosphorylation of each downstream target protein was analyzed. Western blot analysis revealed a significant increase in the phosphorylation of Akt, a target of mTORC2, whereas the phosphorylation of S6K, a target of mTORC1, was not detected (FIGs.18C-18D and FIG. 19B). Thus, C12-2aNP likely stimulates cell metabolism through the mTORC2 signaling pathway rather than mTORC1. Next, the transcriptome of C12-2aNP-treated BMDCs was evaluated using RNA-Seq, and it was found that the expression of genes involved in glycolysis was significantly up- regulated (FIG.18E). For example, compared to the C12-NP-treated group, the C12-2aNP- treated group showed a 2.90-fold higher mRNA expression of glucose transporter-1 (Slc2a1). In addition, the expressions of hexokinase (Hk2), phosphofructokinase (Pfkl), and pyruvate kinase (Pkm), which are the key enzymes of glycolysis, were 3.03-, 2.09-, and 1.62-fold increased in the C12-2aNP-treated group compared to the C12-NP treated group. Gene set enrichment analysis of the C12-2aNP-treated group supported this finding that glycolysis genes are over-represented compared to the untreated group, showing a normalized enrichment score (NES) of 1.66 with a false discovery rate (FDR) q value of 0.042 (FIG. 18F). In contrast, the C12-2NP-treated group showed an NES of 1.21 with an FDR q value of 0.339 (FIG.19C). Additionally, glucose metabolism-related genes were found to be up- regulated by C12-2aNP through pathway enrichment analysis. In this analysis, the fold enrichment of the glycolysis pathway gene set was 3.1 with an enrichment FDR value of 6.0E-6, indicating a significant increase in glycolysis level. The pathway enrichment analysis also revealed that C12-2aNP enhances glucose metabolism-related gene expression broadly by affecting glycan, glycosaminoglycan, and other sugar-related metabolic pathways (FIG. 18G). As a result of the upregulation of glycolysis related genes, C12-2aNP-treated BMDCs exhibited elevated glycolysis rates. The C12-2aNP-treated group showed higher extracellular acidification rate (ECAR) upon glucose and oligomycin treatment than ECAR of the untreated and C12-NP (FIG.18H). Basal glycolysis of C12-2aNP was maintained significantly higher, showing 3.10-fold higher normalized ECAR than that of C12-NP-treated group (FIG.18J). Consistently, maximal glycolytic capacity of C12-2aNP-treated group was significantly enhanced, showing 1.73-fold higher normalized ECAR than that of C12-NP- treated group. On the other hand, aerobic respiration, another major energy-producing pathway related to oxidative phosphorylation, was not altered by C12-2aNP (FIG.18I). When mitochondrial respiration was assessed, the basal oxygen consumption rate (OCR) of the - 133 - 53224792.3 Attorney Docket No.0466483-7483WO1 C12-2aNP-treated group remained significantly lower than that of the untreated and C12-NP- treated groups, indicating that cellular energy metabolism is directed toward glycolysis (FIG. 19D). However, there was no significant difference between the untreated and LNP-treated groups in maximal respiration upon oligomycin and FCCP treatment, indicating no change in mitochondrial function for oxidative phosphorylation. As a result, stimulation of glycolysis by C12-2aNP treatment placed BMDCs in a metabolically glycolytic state while C12-NP- treated cells remained in a metabolically aerobic state similar to untreated BMDCs (FIG. 18K). Consequently, C12-2aNP-treated BMDCs exhibited higher ATP production compared to the untreated and C12-NP-treated groups (FIG.18L). While the C12-2aNP-treated group showed a glycolytic ATP production rate of 22.5 pmol / min / 1000 cells and a mitochondrial ATP production rate of 3.8 pmol / min / 1000 cells, the untreated and C12-NP-treated group showed glycolytic ATP production rates of 14.6 and 17.2 pmol / min / 1000 cells and mitochondrial ATP production rates of 8.6 and 7.3 pmol / min / 1000, respectively. Example 11: C12-2aNP-mediated vaccination elicits a robust immune response in SARS-CoV-2 model Encouraged by the enhanced mRNA transfection and metabolic-modulating properties of C12-2aNP, its efficacy as an mRNA vaccine was evaluated in a SARS-CoV-2 model. For immunization, the mRNA encoding the receptor binding domain (RBD) of the SARS-CoV-2 spike protein (mRBD) was loaded into LNPs and intramuscularly injected to mice. An LNP based on the ALC-0315 ionizable lipid, which was used in an FDA-approved COVID-19 mRNA vaccine, was included as a positive control. After the prime and boost injection of mRBD-loaded C12-2aNP, significant induction of humoral immune responses was detected. In blood serum, the antibody titer of anti-RBD IgG in the C12-2aNP-treated group was comparable to the titer in the ALC-0315-treated group but significantly higher than the titer in the C12-NP-treated group (FIG.20B). Consistently, the induction of antigen-specific B cells was also detected in secondary lymphoid organs. In the spleen, the population of RBD- specific B cells was significantly increased in both the ALC-0315-treated group and the C12- 2aNP-treated group. In particular, the C12-2aNP-treated group showed a 5.68-fold increase in RBD-specific B cells (FIGs.20C-20D). As both the C12-2aNP and ALC-0315-treated groups induced high levels of RBD- specific antibody production, the antibody provided a neutralization effect against SARS- CoV-2 pseudovirus infection. Incubation of the serum from the C12-2aNP or of ALC-0315- treated groups with the pseudovirus significantly interfered with the infection of hACE- - 134 - 53224792.3 Attorney Docket No.0466483-7483WO1 expressing cells, whereas the serum from the C12-NP-treated group exerted a moderate level of interference (FIG.20E). The average 50 % neutralization titers of the C12-2aNP-treated group was 1.66 × 103, which was comparable to that of the ALC-0315-treated group and significantly higher than that of the C12-NP-treated group, which showed an average titer of 0.11 × 103(FIG.20F). Along with the humoral immune responses, C12-2aNP vaccination also induced robust cellular immune responses. When RBD-specific T cells in the spleen were analyzed by IFN-g ELISpot, significantly higher spot numbers were counted in the C12-2aNP- and ALC- 0315-treated groups (FIG.20G and FIG.21E). Specifically, the number of IFN-g secreting cells in the spleen of the C12-2aNP-treated group was 2.39-fold higher than that of the C12- NP-treated group, indicating a stronger cellular immune response against the RBD antigen. Likewise, intracellular cytokine analysis also revealed that higher levels of proinflammatory cytokines, IFN-g and TNF-a, were induced in CD8 T cells in the C12-2aNP-treated group compared to those in the C12-NP-treated group (FIG.20H and FIG.21A). Establishment of immunity against the RBD antigen was also visualized by immunofluorescence staining (FIG.20I). It was confirmed that C12-2aNP immunization robustly induced germinal center (GC) formation, characterized by clustered GL-7+GC B cells and CD21+follicular dendritic cells. While the spleen tissue of the C12-2aNP-treated group showed clusters of these two types of cells, indicating GC formation, the cells did not appear to be clustered together in the spleen of the C12-NP-treated group, indicating inefficient GC formation. In addition, when GC formation was quantitatively evaluated by flow cytometry, a notable induction of antigen-specific GC B cells was detected in the spleen of the C12-2aNP-treated group, significantly higher than the group treated with ALC-0315. Specifically, the population of RBD-specific GC B cells in the C12-2aN-treated group was 6.77 ± 2.27 %, which were 4.23-fold higher than that of the ALC-0315, which had a population of 1.59 ± 0.57 % (FIG.20J). Since the GC plays an important role in the establishment of immunological memory, subsequent memory immunity formation was further evaluated by flow cytometry. In the spleen of immunized mice, robust induction of RBD-specific memory B cells was confirmed in the C12-2aNP-treated group, showing a 1.45-fold higher population than that in the group treated with ALC-0315 (FIG.20K). Regarding follicular help T cells (TFH), which are specialized cells that support B cells in forming GC memory B cells, C12-2aNP- mediated vaccination induced an increase in the cell population. While C12-2aNP-and ALC-0315 showed similar levels, the population in the C12-2aNP group was 1.75-fold higher than that - 135 - 53224792.3 Attorney Docket No.0466483-7483WO1 in the group treated with C12-NP (FIG.20L and FIG.21B). Furthermore, vaccination induced not only memory B cells but also memory T cells in the spleen. C12-2aNP induced the highest level of CD8+effector memory T cells (TEM) at 86.64 ± 4.62 %, which was higher than that of the ALC-0315-treated group which had 82.24 ± 2.78 % and significantly higher than that of the C12-NP treated group which had 32.88 ± 6.30 % (FIG.20M and FIG.21C). For long term memory evaluation, long-lived plasma cells in the bone marrow were analyzed 3 months after the boost injection. When the number of RBD-specific antibody- secreting cells in the bone marrow was visualized by ELISpot, a significantly higher number of spots were detected in the C12-2aNP-treated group, with 4.68- and 1.46-fold higher antibody secreting cells compared to the groups treated with C12-NP and ALC-0315, respectively (FIG.20N and FIG.21E). Altogether, C12-2aNP-mediated vaccination elicits a robust humoral and cellular immune response, including long-term memory, in the SARS- CoV-2 vaccine model. Example 12: C12-2aNP provides protection against tumors in a cancer vaccine model The efficacy of C12-2aNP as an mRNA vaccine was also evaluated in an ovalbumin (OVA) antigen-based cancer vaccine model. For the preventive vaccine model, OVA encoding mRNA (mOVA) was loaded into LNPs and intramuscularly injected to mice, followed by subcutaneous inoculation of OVA-expressing B16 melanoma (B16-OVA) cells (FIG.22A). Consistent with the results of the SARS-CoV-2 vaccine model, the prime and boost injection of mOVA-loaded C12-2aNP induced robust humoral and cellular immune responses. A significantly higher level of OVA-specific antibody was induced in both ALC- 0315 and C12-2aNP groups, with the titer of C12-2aNP-treated group being significantly higher than that in group treated with C12-NP (FIG.22B). Likewise, a higher number of IFN- g-secreting cells upon OVA antigen stimulation was found in the spleen of the C12-2aNP- treated group compared to the group treated with C12-NP (FIG.22C and FIG.23A). When the population of OVA-specific CD8+T cells was analyzed by flow cytometry, a significantly higher level of the T cells was detected in the spleen of the C12-2aNP-treated group, showing 4.47- and 1.31-fold higher populations than that in C12-NP and ALC-0315-treated groups, respectively (FIG.22D-22E). The robust humoral and cellular immune responses provided the immunized mice with protection against a tumor challenge. When B16-OVA was inoculated to immunized mice, three of six mice vaccinated with C12-2aNP or ACL-0315 showed a complete response, while all of the C12-N-treated mice showed tumor growth (FIG.23B). Tumor - 136 - 53224792.3 Attorney Docket No.0466483-7483WO1 volume monitoring revealed significant tumor growth inhibition in the C12-2aNP or ALC- 0315 groups. In particular, the tumor volume of the C12-2aNP-treated group measured on day 23 was 30.3 ± 44.2 mm3, whereas for the C12-NP-treated group it was 311.9 ± 147.7 mm3(FIG.22F). The potency of C12-2aNP as a cancer vaccine was also demonstrated in a therapeutic vaccine model, where prime and boost immunizations were administered after tumor inoculation (FIG.22G). Similar to the preventive vaccine model, both the C12-2aNP and ALC-0315-treated groups induced comparable anti-tumor efficacy, which was significantly more potent than the C12-NP-treated group. Specifically, the tumor volume in the C12-2aNP- treated group was 402.66 ± 165.8 mm3, which was significantly lower than that in the C12- NP-treated group, which was 718.6 ± 333.9 mm3(FIG.22H and FIG.23C). The tumor growth inhibition led to longer survival rates. While the C12-NP-treated group showed a maximum survival time increase by up to 6 days compared to the untreated group, C12-2aNP vaccination resulted in significant extension of the maximum survival time by up to 18 days (FIG.22I). Stronger immune responses following the C12-2aNP vaccination resulted in a higher level of tumor-infiltrating lymphocytes. Immunofluorescence staining revealed active infiltration of IFN-g expressing CD8+T cells in the tumor of the C12-2aNP-treated group. In contrast, the C12-NP-treated group not only had a lower density of infiltrating cells but also exhibited a lower expression ratio of IFN-γ (FIG.22J). Flow cytometry analysis showed that the number of CD8+T cells infiltrating the tumor in the C12-2aNP-treated group was 3.09- fold higher than that in the C12-NP-treated group (FIG.22K and FIG.23D). Additionally, the proportion of infiltrating cells expressing IFN-γ was also higher in the C12-2aNP-treated group compared to the C12-NP-treated group (FIG.22L and FIG.23E). Sequence Listing SEQ ID NO:1 OVASIINFEKL Enumerated Embodiments The following exemplary embodiments are provided, the numbering of which is not to be construed as designating levels of importance: Embodiment 1 provides a compound of formula (I), or a salt, stereoisomer, or isotopologue thereof: - 137 - 53224792.3 Attorney Docket No.0466483-7483WO1 (I), wherein: A is selected from the group consisting of ; each occurrence of L1, if from the group consisting of -(optionally substituted C1- , C2-C12alkenylenyl)-, - (optionally substituted C1-C12 alkynylenyl)-, -(optionally substituted C1-C12 heteroalkylenyl)- , -(optionally substituted C3-C8cycloalkylenyl)-, -(optionally substituted C2-C8heterocyloalkylenyl)-, -(optionally substituted C6-C10 arylenyl)-, -(optionally substituted C2- C8heteroarylenyl)-, -(optionally substituted C1-C12alkylenyl)-X-, -(optionally substituted C2- C12 alkenylenyl)-X-, -(optionally substituted C1-C12 alkynylenyl)-X-, -(optionally substituted C1-C12heteroalkylenyl)-X-, -(optionally substituted C3-C8cycloalkylenyl)-X-, -(optionally substituted C2-C8 heterocyloalkylenyl)-X-, -(optionally substituted C6-C10 arylenyl)-X-, and - (optionally substituted C2-C8heteroarylenyl)-X-; each occurrence of X, if present, is independently selected from the group consisting of -N(R1e)-, -N(RA)-, and -O-; each occurrence of R1a, R1b, R1c, R1d, and R1e, if present, is independently , wherein: R1cand R1dcan combine with the atoms to which they are bound to form an optionally substituted C2-C8 heterocycloalkyl, or one of R1cand R1dcan combine with one occurrence of L1to form an optionally substituted C2-C8 heterocycloalkyl; R2is selected from the group consisting of optionally substituted C1-C6alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C6heteroalkyl, optionally substituted C3-C6heteroalkenyl, optionally substituted C2-C8 heterocycloalkyl, and N(RA)(RB); each occurrence of L2is independently selected from the group consisting of - (optionally substituted C1-C6 alkylenyl)-, -C(=O)-, -O-, and -N(RA)-; each occurrence of R3is independently selected from the group consisting of optionally substituted C1-C24alkyl and optionally substituted C1-C24heteroalkyl; each occurrence of Ar is independently selected from the group consisting of - - 138 - 53224792.3 Attorney Docket No.0466483-7483WO1 (optionally substituted C6-C10 heteroarylenyl)- and -(optionally substituted C2-C8 heteroarylenyl)-; m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; and each occurrence of RAand RBis independently selected from the group consisting of H, optionally substituted C1-C6alkyl, optionally substituted C2-C6alkenyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C6 heteroalkyl, optionally substituted C3-C6heteroalkenyl, optionally substituted C2-C8heterocycloalkyl, optionally substituted C6- C10 aryl, and optionally substituted C2-C8 heteroaryl. Embodiment 2 provides the compound of Embodiment 1, wherein each occurrence of L1is independently selected from the group consisting of -(CH2)1-5-, -(CH2)1-5N(R1e)-, and . 3 provides the compound of Embodiment 1 or 2, wherein -(L1)m- is selected from the group consisting of: . 1-3, wherein the compound of formula (I) is selected from the group consisting of: each occurrence of L2is independently selected from the group consisting of -(CH2)1-5-, -CH(OH)- , and -O-. Embodiment 6 provides the compound of any one of Embodiments 1-5, wherein each occurrence of R1a, R1b, R1c, R1d, and R1e, if , - 139 - 53224792.3 Attorney Docket No.0466483-7483WO1 wherein: each occurrence of R4is independently selected from the group consisting of optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C3-C8cycloalkyl, optionally substituted C2-C6heteroalkyl, optionally substituted C3-C6heteroalkenyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C6-C10 aryl, optionally substituted C2-C8heteroaryl, ORA, N(RA)(RB), halogen, CN, NO2, C(=O)RA, C(=O)ORA, C(=O)N(RA)(RB), S(=O)2RA, S(=O)2ORA, and S(=O)2N(RA)(RB), wherein two vicinal R4moieties can combine with the carbon atoms to which they are bound to form an optionally substituted C3-C8 cycloalkyl, C2- C8heterocycloalkyl, C6-C10aryl, or C2-C8heteroaryl; and o is an integer selected from the group consisting of 0, 1, 2, 3, and 4. Embodiment 7 provides the compound of any one of Embodiments 1-6, wherein each occurrence of -(L2)n- is independently selected from the group consisting of: . Embodiment 1-7, wherein each occurrence of R1a, and if present, is independently selected from the group consisting of: OH SS R3, . occurrence of R3is independently optionally substituted C4-C12alkyl. Embodiment 10 provides the compound of any one of Embodiments 1-9, wherein each occurrence of R3is independently selected from the group consisting of n-butyl, n- pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, n-dodecyl. Embodiment 11 provides the compound of any one of Embodiments 1-10, wherein - 140 - 53224792.3 Attorney Docket No.0466483-7483WO1 each occurrence of R1a, R1b, R1c, R1d, and R1e, if present, is independently selected from the group consisting of: , , - 141 - 53224792.3 Attorney Docket No.0466483-7483WO1 . the compound is selected from the group consisting of: 3,3'-((2-(4-(2-((2-(bis(3-(2-(hexyldisulfaneyl)phenoxy)-2- h l i thl 2 h li lf l h 2 - 142 - 53224792.3 Attorney Docket No.0466483-7483WO1 hydroxypropyl)amino)ethyl)piperazin-1-yl)ethyl)azanediyl)bis(1-(2- (decyldisulfaneyl)phenoxy)propan-2-ol); 33'-((2-(4-(2-((2-(bi(3-(2-(dd ldi lf n l)hnx)-2- 1- - 143 - 53224792.3 Attorney Docket No.0466483-7483WO1 C8mPhE- 3,3'-((2-(4-(2-hydroxy-3-(3-(octyldisulfaneyl)phenoxy)propyl)piperazin-1- 318 yl)ethyl)azanediyl)bis(1-(3-(octyldisulfaneyl)phenoxy)propan-2-ol); C10mPhE 33'-((2-(4-(3-(3-(d ldi lf n l)hnx)-2-hdrx r l)i rzin- 1- ); ); ); - ); ); ); - ); ); ); - (a) at least one ionizable lipid comprising the compound of any one of Embodiments 1-13; (b) at least one neutral lipid; - 144 - 53224792.3 Attorney Docket No.0466483-7483WO1 (c) at least one cholesterol lipid and / or a modified derivative thereof; and (d) at least one polymer-conjugated lipid and / or a modified derivative thereof. Embodiment 14 provides the LNP of Embodiment 13, wherein the at least one ionizable lipid compound comprises about 10 mol% to about 90 mol% of the LNP, optionally wherein the at least one ionizable lipid compound comprises about 35 mol% of the LNP. Embodiment 15 provides the LNP of any one of Embodiments 13 or 14, wherein the at least one neutral lipid comprises about 1 mol% to about 40 mol% of the LNP, optionally wherein the at least one neutral lipid comprises about 16 mol% of the LNP. Embodiment 16 provides the LNP of any one of Embodiments 13-15, wherein the neutral lipid comprises or consists essentially of at least one neutral lipid selected from the group consisting of dioleoylphosphatidylethanolamine (DOPE), distearoylphosphatidylcholine (DSPC), and dioleoylphosphatidylcholine (DOPC), optionally wherein the neutral lipid comprises or consists essentially of dioleoylphosphatidylethanolamine (DOPE). Embodiment 17 provides the LNP of any one of Embodiments 13-16, wherein the at least one cholesterol lipid and / or modified derivative thereof comprises about 20 mol% to about 75 mol% of the LNP, optionally wherein the at least one cholesterol lipid and / or modified derivative thereof comprises about 46.5 mol% of the LNP. Embodiment 18 provides the LNP of any one of Embodiments 13-17, wherein the at least one cholesterol lipid and / or modified derivative thereof comprises or consists essentially of cholesterol. Embodiment 19 provides the LNP of any one of Embodiments 13-18, wherein the at least one polymer-conjugated lipid comprises about 0.1 mol% to about 15 mol% of the LNP, optionally wherein the at least one polymer-conjugated lipid comprises about 2.5 mol%. Embodiment 20 provides the LNP of any one of Embodiments 13-19, wherein the at least one polymer-conjugated lipid comprises or consists essentially of 1,2-dimyristoyl-sn- glycero-3-phosphoethanolamine-N-[methoxy(polyethyleneglycol)-2000] (C14PEG2K). Embodiment 21 provides the LNP of any one of Embodiments 13-20, wherein the LNP has a molar ratio of (a) : (b) : (c) : (d) of about 35:16:46.5:2.5. Embodiment 22 provides the LNP of any one of Embodiments 13-21, wherein the LNP further comprises at least one cargo molecule, optionally wherein the at least one cargo molecule comprises or consists of a therapeutic cargo molecule. Embodiment 23 provides the LNP of Embodiment 22, wherein the cargo molecule is at least one selected from the group consisting of a nucleic acid, small molecule, protein, - 145 - 53224792.3 Attorney Docket No.0466483-7483WO1 therapeutic agent, antibody, and any combinations thereof. Embodiment 24 provides the LNP of Embodiment 22 or 23, wherein the cargo molecule comprises a nucleic acid. Embodiment 25 provides the LNP of Embodiment 24, wherein the nucleic acid is DNA or RNA. Embodiment 26 provides the LNP of Embodiment 24 or 25, wherein the nucleic acid is selected from the group consisting of mRNA, cDNA, pDNA, microRNA, sgRNA, siRNA, modified RNA, antagomir, antisense molecule, and any combinations thereof. Embodiment 27 provides the LNP of Embodiment 26, wherein the mRNA encodes an enzyme, receptor, or antigen binding domain. Embodiment 28 provides the LNP of Embodiment 26, wherein the mRNA encodes a clustered regularly interspaced short palindrome repeats (CRISPR) associated protein, optionally wherein the CRISPR associated protein is Cas9. Embodiment 29 provides the LNP of Embodiment 28, wherein the nucleic acid cargo further comprises sgRNA. Embodiment 30 provides a pharmaceutical composition comprising the lipid nanoparticle (LNP) of any one of Embodiments 13-28 and at least one pharmaceutically acceptable carrier. Embodiment 31 provides the pharmaceutical composition of Embodiment 30, wherein the composition is formulated for use as a vaccine. Embodiment 32 provides the pharmaceutical composition of Embodiment 31, wherein the composition further comprises at least one adjuvant. Embodiment 33 provides a method of treating, preventing, and / or ameliorating a disease or infection in a subject, the method comprising administering to the subject at least one lipid nanoparticle (LNP) of any one of Embodiments 23-29 or the pharmaceutical composition of any one of Embodiments 30-32. Embodiment 34 provides the method of Embodiment 33, wherein the disease or infection is at least one selected from the group consisting of cancer, a viral infection, an immunomodulatory disease, and a lymphatic disease. Embodiment 35 provides a method of delivering at least one therapeutic cargo molecule to a lymph node of a subject, the method comprising administering to the subject at least one lipid nanoparticle (LNP) of any one of Embodiments 23-29 or the pharmaceutical composition of any one of Embodiments 30-32. Embodiment 36 provides the method of Embodiment 35, wherein the lymph node is a - 146 - 53224792.3 Attorney Docket No.0466483-7483WO1 pelvic lymph node, optionally wherein the pelvic lymph node is an inguinal lymph node or an iliac lymph node. Embodiment 37 provides the method of any one of Embodiments 33-36, wherein the subject is a mammal. Embodiment 38 provides the method of Embodiment 37, wherein the mammal is a human. Embodiment 39 provides a compound of formula (II), or a salt, stereoisomer, or isotopologue thereof: , wherein: R1a, R1b, and independently a polyamine moiety, wherein is substituted with at least one optionally substituted C1-C20 alkyl, optionally substituted C1-C20 heteroalkyl, optionally substituted C2-C20 alkenyl, optionally substituted C2-C20 heteroalkyenyl, optionally substituted C2-C20alkynyl, and optionally substituted C2-C20heteroalkynyl; R2a, R2b, and each occurrence of R2c, if present, are each independently selected from the and optionally substituted C1-C6alkyl; R3a, R3b, and each occurrence of R3c, if present, are each independently selected from the group consisting of H and optionally substituted C1-C6alkyl; each occurrence of R4aand R4bis independently selected from the group consisting of H, optionally substituted C1-C6alkyl, optionally substituted C1-C6heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C6-C10aryl, and optionally substituted C2-C8heteroaryl; each occurrence of R5aand R5bis independently selected from the group consisting of H, optionally substituted C1-C6alkyl, optionally substituted C1-C6heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C6-C10aryl, and optionally substituted C2-C8heteroaryl; each occurrence of R6is ; each occurrence of R7aand R7b, if selected from the group - 147 - 53224792.3 Attorney Docket No.0466483-7483WO1 consisting of H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C8cycloalkyl, optionally substituted C2-C8heterocycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted C2-C8 heteroaryl; m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each occurrence of n is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; o is 0, 1, 2, 3, 4, or 5; and each occurrence of p is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. Embodiment 40 provides the compound of Embodiment 39, wherein at least one of the following applies: (a) R2a, R2b, and each occurrence of R2c, if present, are each independently H; (b) R3a, R3b, and each occurrence of R3c, if present, are each independently H; (c) each occurrence of R4aand R4bis independently H; (d) each occurrence of R5aand R5bis independently H; and (e) each occurrence of R7aand R7bis independently H. Embodiment 41 provides the compound of Embodiment 39 or 40, wherein the compound of formula (II) is selected from the group consisting of: wherein R1a, R1b, and each occurrence of R1c, if present, are each independently , wherein: each occurrence of L1, if present, is independently selected from the group consisting of -(optionally substituted C1-C12alkylenyl)-, -(optionally substituted C2-C12alkenylenyl)-, - - 148 - 53224792.3 Attorney Docket No.0466483-7483WO1 (optionally substituted C1-C12 alkynylenyl)-, -(optionally substituted C1-C12 heteroalkylenyl)- , -(optionally substituted C3-C8cycloalkylenyl)-, -(optionally substituted C2-C8heterocyloalkylenyl)-, -(optionally substituted C6-C10 arylenyl)-, -(optionally substituted C2- C8heteroarylenyl)-, -(optionally substituted C1-C12alkylenyl)-X-, -(optionally substituted C2- C12 alkenylenyl)-X-, -(optionally substituted C1-C12 alkynylenyl)-X-, -(optionally substituted C1-C12heteroalkylenyl)-X-, -(optionally substituted C3-C8cycloalkylenyl)-X-, -(optionally substituted C2-C8 heterocyloalkylenyl)-X-, -(optionally substituted C6-C10 arylenyl)-X-, - (optionally substituted C2-C8heteroarylenyl)-X-, and -X-; each occurrence of X, if present, is independently selected from the group consisting of -N(R8c)-, -N(RA)-, -O-, and -S-; each occurrence of R8a, R8b, and R8c, if present, is independently selected from the group consisting of optionally substituted C1-C20alkyl, optionally substituted C1-C20heteroalkyl, optionally substituted C2-C20 alkenyl, optionally substituted C2-C20 heteroalkyenyl, optionally substituted C2-C20alkynyl, and optionally substituted C2-C20heteroalkynyl, wherein one of R8aand R8bcan combine with one occurrence of L1to form an optionally substituted C2-C8 heterocycloalkyl; each occurrence of RAand RBis independently selected from the group consisting of H and optionally substituted C1-C6alkyl; and each occurrence of q is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. Embodiment 43 provides the compound of Embodiment 42, wherein each occurrence of L1is independently selected from the group consisting of -(CH2)1-5-, -(CH2)1-5CH(ORA)-, - -(L1)p- is selected from the group consisting of: , - 149 - 53224792.3 Attorney Docket No.0466483-7483WO1 , , R1a, R1b, and each occurrence of R1c, if present, is independently selected from the group consisting of: , , - 150 - 53224792.3 Attorney Docket No.0466483-7483WO1 , each occurrence of R8a, R8b, and R8cis independently selected from the group consisting of - (CH2)CH(OH)(optionally substituted C3-C20 alkyl). Embodiment 47 provides the compound of any one of Embodiments 42-46, wherein each R8a, R8b, and R8cis independently selected from the group consisting of - (CH2)CH(OH)(CH2)3CH3, -(CH2)CH(OH)(CH2)3CH3, -(CH2)CH(OH)(CH2)4CH3, - (CH2)CH(OH)(CH2)5CH3, -(CH2)CH(OH)(CH2)6CH3, -(CH2)CH(OH)(CH2)7CH3, - (CH2)CH(OH)(CH2)8CH3, -(CH2)CH(OH)(CH2)9CH3, -(CH2)CH(OH)(CH2)10CH3, - (CH2)CH(OH)(CH2)11CH3, -(CH2)CH(OH)(CH2)12CH3, -(CH2)CH(OH)(CH2)13CH3, - (CH2)CH(OH)(CH2)14CH3, -(CH2)CH(OH)(CH2)15CH3, -(CH2)CH(OH)(CH2)16CH3, - - 151 - 53224792.3 Attorney Docket No.0466483-7483WO1 (CH2)CH(OH)(CH2)17CH3, -(CH2)CH(OH)(CH2)18CH3, -(CH2)CH(OH)(CH2)19CH3, and - (CH2)CH(OH)(CH2)20CH3. Embodiment 48 provides the compound of any one of Embodiments 39-47, wherein R1a, R1b, and each occurrence of R1c, if present, is independently selected from the group consisting of: , , - 152 - 53224792.3 Attorney Docket No.0466483-7483WO1 5 , - 153 - 53224792.3 Attorney Docket No.0466483-7483WO1 39-48, wherein the compound of formula (II) is: yl)propyl)adipimidamide, (C12-2aN). Embodiment 50 provides a lipid nanoparticle (LNP) comprising: (a) at least one ionizable lipid comprising the compound of any one of Embodiments 1-11; (b) at least one neutral lipid; (c) at least one cholesterol lipid and / or a modified derivative thereof; and (d) at least one polymer-conjugated lipid and / or a modified derivative thereof. Embodiment 51 provides the LNP of Embodiment 50, wherein the at least one ionizable lipid compound comprises about 10 mol% to about 90 mol% of the LNP, optionally wherein the at least one ionizable lipid compound comprises about 57.1 mol% or about 50 mol% of the LNP. Embodiment 52 provides the LNP of Embodiment 50 or 51, wherein the at least ionizable lipid compound comprises or consists essentially of: - 154 - 53224792.3 Attorney Docket No.0466483-7483WO1 (C12-2aN). Embodiment 53 provides the LNP of any one of Embodiments 50-52, wherein the at least one neutral lipid comprises about 1 mol% to about 40 mol% of the LNP, optionally wherein the at least one neutral lipid comprises about 10.2 mol% or about 10.0 mol% of the LNP. Embodiment 54 provides the LNP of any one of Embodiments 50-53, wherein the neutral lipid comprises or consists essentially of at least one neutral lipid selected from the group consisting of dioleoylphosphatidylethanolamine (DOPE), distearoylphosphatidylcholine (DSPC), and dioleoylphosphatidylcholine (DOPC), optionally wherein the neutral lipid comprises or consists essentially of dioleoylphosphatidylethanolamine (DOPE). Embodiment 55 provides the LNP of any one of Embodiments 50-54, wherein the at least one cholesterol lipid and / or modified derivative thereof comprises about 20 mol% to about 75 mol% of the LNP, optionally wherein the at least one cholesterol lipid and / or modified derivative thereof comprises about 31.1 mol% or about 38.5 mol% of the LNP. Embodiment 56 provides the LNP of any one of Embodiments 50-55, wherein the at least one cholesterol lipid and / or modified derivative thereof comprises or consists essentially of cholesterol. Embodiment 57 provides the LNP of any one of Embodiments 50-56, wherein the at least one polymer-conjugated lipid comprises about 0.1 mol% to about 15 mol% of the LNP, optionally wherein the at least one polymer-conjugated lipid comprises about 1.7 mol% or about 1.5 mol% of the LNP. Embodiment 58 provides the LNP of any one of Embodiments 50-57, wherein the at least one polymer-conjugated lipid comprises or consists essentially of 1,2-dimyristoyl-sn- glycero-3-phosphoethanolamine-N-[methoxy(polyethyleneglycol)-2000] (C14PEG2K). Embodiment 59 provides the LNP of any one of Embodiments 50-58, wherein the LNP has a molar ratio of (a) : (b) : (c) : (d) of about 57.1 : 10.2 : 31.1 : 1.7 or about 50 : 10 : - 155 - 53224792.3 Attorney Docket No.0466483-7483WO1 38.5 : 1.5. Embodiment 60 provides the LNP of any one of Embodiments 50-59, wherein the LNP further comprises at least one cargo molecule, optionally wherein the at least one cargo molecule comprises or consists of a therapeutic cargo molecule. Embodiment 61 provides the LNP of Embodiment 60, wherein the cargo molecule is at least one selected from the group consisting of a nucleic acid, small molecule, protein, therapeutic agent, antibody, and any combinations thereof. Embodiment 62 provides the LNP of Embodiment 60 or 61, wherein the cargo molecule comprises a nucleic acid. Embodiment 63 provides the LNP of Embodiment 62, wherein the nucleic acid is DNA or RNA. Embodiment 64 provides the LNP of Embodiment 62 or 63, wherein the nucleic acid is selected from the group consisting of mRNA, cDNA, pDNA, microRNA, sgRNA, siRNA, modified RNA, antagomir, antisense molecule, and any combinations thereof. Embodiment 65 provides the LNP of Embodiment 64, wherein the mRNA encodes an enzyme, receptor, or antigen binding domain. Embodiment 66 provides the LNP of Embodiment 64, wherein the mRNA encodes a clustered regularly interspaced short palindrome repeats (CRISPR) associated protein, optionally wherein the CRISPR associated protein is Cas9. Embodiment 67 provides the LNP of Embodiment 66, wherein the nucleic acid cargo further comprises sgRNA. Embodiment 68 provides a pharmaceutical composition comprising the lipid nanoparticle (LNP) of any one of Embodiments 50-67 and at least one pharmaceutically acceptable carrier. Embodiment 69 provides a method of treating, preventing, and / or ameliorating a disease in a subject, the method comprising administering to the subject at least one lipid nanoparticle (LNP) of any one of Embodiments 50-67 or the pharmaceutical composition of Embodiment 68. Embodiment 70 provides the method of Embodiment 69, wherein the disease is at least one selected from the group consisting of cancer, immune-mediated diseases, cardiovascular disease, and a metabolic disease. Embodiment 71 provides a method of delivering at least one therapeutic cargo molecule to a dendritic cell of a subject, the method comprising administering to the subject at least one lipid nanoparticle (LNP) of any one of Embodiments 50-67 or the pharmaceutical - 156 - 53224792.3 Attorney Docket No.0466483-7483WO1 composition of Embodiment 68. Embodiment 72 provides the method of Embodiment 71, wherein the therapeutic cargo molecule is mRNA. Embodiment 73 provides the method of Embodiment 72, wherein the LNP stimulates glycolysis in the dendritic cell. Embodiment 74 provides the method of Embodiment 72 or 73, wherein the LNP facilitates endosomal escape. The terms and expressions employed herein are used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the embodiments of the present application. Thus, it should be understood that although the present application describes specific embodiments and optional features, modification and variation of the compositions, methods, and concepts herein disclosed may be resorted to by those of ordinary skill in the art, and that such modifications and variations are considered to be within the scope of embodiments of the present application. - 157 - 53224792.3

Claims

Attorney Docket No.0466483-7483WO1 CLAIMS What is claimed is:

1. A compound of formula (I), or a salt, stereoisomer, or isotopologue thereof: (I), wherein: A is selected from the group consisting of ; each occurrence of L1, if from the group consisting of -(optionally substituted C -1 -, - C2-C12 alkenylenyl)-, - (optionally substituted C1-C12alkynylenyl)-, -(optionally substituted C1-C12heteroalkylenyl)- , -(optionally substituted C3-C8 cycloalkylenyl)-, -(optionally substituted C2-C8 heterocyloalkylenyl)-, -(optionally substituted C6-C10arylenyl)-, -(optionally substituted C2- C8 heteroarylenyl)-, -(optionally substituted C1-C12 alkylenyl)-X-, -(optionally substituted C2- C12alkenylenyl)-X-, -(optionally substituted C1-C12alkynylenyl)-X-, -(optionally substituted C1-C12 heteroalkylenyl)-X-, -(optionally substituted C3-C8 cycloalkylenyl)-X-, -(optionally substituted C2-C8heterocyloalkylenyl)-X-, -(optionally substituted C6-C10arylenyl)-X-, and - (optionally substituted C2-C8 heteroarylenyl)-X-; each occurrence of X, if present, is independently selected from the group consisting of -N(R1e)-, -N(RA)-, and -O-; each occurrence of R1a, R1b, R1c, R1d, and R1e, if present, is independently , wherein: R1cand R1dcan combine with the atoms to which they are bound to form an optionally substituted C2-C8heterocycloalkyl, or one of R1cand R1dcan combine with one occurrence of L1to form an optionally substituted C2-C8heterocycloalkyl; R2is selected from the group consisting of optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C6 heteroalkyl, optionally substituted C3-C6 heteroalkenyl, optionally substituted C2-C8 heterocycloalkyl, and N(RA)(RB); each occurrence of L2is independently selected from the group consisting of - (optionally substituted C1-C6 alkylenyl)-, -C(=O)-, -O-, and -N(RA)-; - 158 - 53224792.3Attorney Docket No.0466483-7483WO1 each occurrence of R3is independently selected from the group consisting of optionally substituted C1-C24alkyl and optionally substituted C1-C24heteroalkyl; each occurrence of Ar is independently selected from the group consisting of - (optionally substituted C6-C10heteroarylenyl)- and -(optionally substituted C2-C8heteroarylenyl)-; m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; and each occurrence of RAand RBis independently selected from the group consisting of H, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C3-C8cycloalkyl, optionally substituted C2-C6heteroalkyl, optionally substituted C3-C6 heteroalkenyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C6- C10aryl, and optionally substituted C2-C8heteroaryl.

2. The compound of claim 1, wherein each occurrence of L1is independently selected from the group consisting of -(CH2)1-5-, -(CH2)1-5N(R1e)-, and .

3. The compound of claim 1 or 2, wherein -(L1)m- is selected from the group consisting of:.

4. The compound of any one of claims 1-3, wherein the compound of formula (I) is selected from the group consisting of:

5. The compound of any one of claims 1-4, wherein each occurrence of L2is - 159 - 53224792.3Attorney Docket No.0466483-7483WO1 independently selected from the group consisting of -(CH2)1-5-, -CH(OH)-, and -O-.

6. The compound of any one of claims 1-5, wherein each occurrence of R1a, R1b, R1c, R1d, and R1e, if present, is , wherein: each occurrence of R4is group consisting ofoptionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C3-C8cycloalkyl, optionally substituted C2-C6heteroalkyl, optionally substituted C3-C6heteroalkenyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C6-C10 aryl, optionally substituted C2-C8heteroaryl, ORA, N(RA)(RB), halogen, CN, NO2, C(=O)RA, C(=O)ORA, C(=O)N(RA)(RB), S(=O)2RA, S(=O)2ORA, and S(=O)2N(RA)(RB), wherein two vicinal R4moieties can combine with the carbon atoms to which they are bound to form an optionally substituted C3-C8 cycloalkyl, C2- C8heterocycloalkyl, C6-C10aryl, or C2-C8heteroaryl; and o is an integer selected from the group consisting of 0, 1, 2, 3, and 4.

7. The compound of any one of claims 1-6, wherein each occurrence of -(L2)n- is independently selected from the group consisting of: .

8. The any one occurrence of R1a, R1b, R1c, R1d, and R1e, if present, is independently selected from the group consisting of: , .- 160 - 53224792.3Attorney Docket No.0466483-7483WO1 9. The compound of any one of claims 1-8, wherein each occurrence of R3is independently optionally substituted C4-C12 alkyl.

10. The compound of any one of claims 1-9, wherein each occurrence of R3is independently selected from the group consisting of n-butyl, n-pentyl, n-hexyl, n-heptyl, n- octyl, n-nonyl, n-decyl, n-undecyl, n-dodecyl.

11. The compound of any one of claims 1-10, wherein each occurrence of R1a, R1b, R1c, R1d, and R1e, if present, is independently selected from the group consisting of: , , , , , , ,- 161 - 53224792.3Attorney Docket No.0466483-7483WO1 .Attorney Docket No.0466483-7483WO1 12. The compound of any one of claims 1-11, wherein the compound is selected from the group consisting of: 3,3'-((2-(4-(2-((2-(bis(3-(2-(hexyldisulfaneyl)phenoxy)-2- C6oPhE-200 hydroxypropyl)amino)ethyl)(3-(2-(hexyldisulfaneyl)phenoxy)-2- h l i thl i i 1 l thl il i 12- 163 - 53224792.3Attorney Docket No.0466483-7483WO1 200 hydroxypropyl)amino)ethyl)(3-(4-(dodecyldisulfaneyl)phenoxy)-2- hydroxypropyl)amino)ethyl)piperazin-1-yl)ethyl)azanediyl)bis(1-(4- (dodecldisulfanel)henox)roan-2-ol) 1- 1- 1- ); ); ); - ); ); ); -- 164 - 53224792.3Attorney Docket No.0466483-7483WO1 diyl))bis(azanetriyl))tetrakis(1-(4-(hexyldisulfaneyl)phenoxy)propan-2-ol); C8pPhE-383 3,3',3'',3'''-((piperazine-1,4-diylbis(propane-3,1- di l))bi ( z n tri l))t tr ki (1-(4-( t ldi lf n l) h n x ) r n-2- l); ); -(a) at least one ionizable lipid comprising the compound of any one of claims 1- 13; (b) at least one neutral lipid; (c) at least one cholesterol lipid and / or a modified derivative thereof; and (d) at least one polymer-conjugated lipid and / or a modified derivative thereof.

14. The LNP of claim 13, wherein the at least one ionizable lipid compound comprises about 10 mol% to about 90 mol% of the LNP, optionally wherein the at least one ionizable lipid compound comprises about 35 mol% of the LNP.

15. The LNP of any one of claims 13 or 14, wherein the at least one neutral lipid comprises about 1 mol% to about 40 mol% of the LNP, optionally wherein the at least one neutral lipid comprises about 16 mol% of the LNP.

16. The LNP of any one of claims 13-15, wherein the neutral lipid comprises or consists essentially of at least one neutral lipid selected from the group consisting of dioleoylphosphatidylethanolamine (DOPE), distearoylphosphatidylcholine (DSPC), and dioleoylphosphatidylcholine (DOPC), optionally wherein the neutral lipid comprises or consists essentially of dioleoylphosphatidylethanolamine (DOPE).

17. The LNP of any one of claims 13-16, wherein the at least one cholesterol lipid and / or modified derivative thereof comprises about 20 mol% to about 75 mol% of the LNP, optionally wherein the at least one cholesterol lipid and / or modified derivative thereof comprises about 46.5 mol% of the LNP. - 165 - 53224792.3Attorney Docket No.0466483-7483WO1 18. The LNP of any one of claims 13-17, wherein the at least one cholesterol lipid and / or modified derivative thereof comprises or consists essentially of cholesterol.

19. The LNP of any one of claims 13-18, wherein the at least one polymer-conjugated lipid comprises about 0.1 mol% to about 15 mol% of the LNP, optionally wherein the at least one polymer-conjugated lipid comprises about 2.5 mol%.

20. The LNP of any one of claims 13-19, wherein the at least one polymer-conjugated lipid comprises or consists essentially of 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine- N-[methoxy(polyethyleneglycol)-2000] (C14PEG2K).

21. The LNP of any one of claims 13-20, wherein the LNP has a molar ratio of (a) : (b) : (c) : (d) of about 35:16:46.5:2.

5.

22. The LNP of any one of claims 13-21, wherein the LNP further comprises at least one cargo molecule, optionally wherein the at least one cargo molecule comprises or consists of a therapeutic cargo molecule.

23. The LNP of claim 22, wherein the cargo molecule is at least one selected from the group consisting of a nucleic acid, small molecule, protein, therapeutic agent, antibody, and any combinations thereof.

24. The LNP of claim 22 or 23, wherein the cargo molecule comprises a nucleic acid.

25. The LNP of claim 24, wherein the nucleic acid is DNA or RNA.

26. The LNP of claim 24 or 25, wherein the nucleic acid is selected from the group consisting of mRNA, cDNA, pDNA, microRNA, sgRNA, siRNA, modified RNA, antagomir, antisense molecule, and any combinations thereof.

27. The LNP of claim 26, wherein the mRNA encodes an enzyme, receptor, or antigen binding domain.

28. The LNP of claim 26, wherein the mRNA encodes a clustered regularly interspaced - 166 - 53224792.3Attorney Docket No.0466483-7483WO1 short palindrome repeats (CRISPR) associated protein, optionally wherein the CRISPR associated protein is Cas9.

29. The LNP of claim 28, wherein the nucleic acid cargo further comprises sgRNA.

30. A pharmaceutical composition comprising the lipid nanoparticle (LNP) of any one of claims 13-28 and at least one pharmaceutically acceptable carrier.

31. The pharmaceutical composition of claim 30, wherein the composition is formulated for use as a vaccine.

32. The pharmaceutical composition of claim 31, wherein the composition further comprises at least one adjuvant.

33. A method of treating, preventing, and / or ameliorating a disease or infection in a subject, the method comprising administering to the subject at least one lipid nanoparticle (LNP) of any one of claims 23-29 or the pharmaceutical composition of any one of claims 30-32.

34. The method of claim 33, wherein the disease or infection is at least one selected from the group consisting of cancer, a viral infection, an immunomodulatory disease, and a lymphatic disease.

35. A method of delivering at least one therapeutic cargo molecule to a lymph node of a subject, the method comprising administering to the subject at least one lipid nanoparticle (LNP) of any one of claims 23-29 or the pharmaceutical composition of any one of claims 30-32.

36. The method of claim 35, wherein the lymph node is a pelvic lymph node, optionally wherein the pelvic lymph node is an inguinal lymph node or an iliac lymph node.

37. The method of any one of claims 33-36, wherein the subject is a mammal.

38. The method of claim 37, wherein the mammal is a human. - 167 - 53224792.3Attorney Docket No.0466483-7483WO1 39. A compound of formula (II), or a salt, stereoisomer, or isotopologue thereof: , wherein: R1a, R1b, anda polyamine moiety, wherein each amine substituent of the polyamine moiety is substituted with at least one optionally substituted C1-C20alkyl, optionally substituted C1-C20heteroalkyl, optionally substituted C2-C20 alkenyl, optionally substituted C2-C20 heteroalkyenyl, optionally substituted C2-C20alkynyl, and optionally substituted C2-C20heteroalkynyl; R2a, R2b, and each occurrence of R2c, if present, are each independently selected from the group consisting of H and optionally substituted C1-C6alkyl; R3a, R3b, and each occurrence of R3c, if present, are each independently selected from the and optionally substituted C1-C6 alkyl;each occurrence of R4aand R4bis independently selected from the group consisting of H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C8cycloalkyl, optionally substituted C2-C8heterocycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted C2-C8 heteroaryl; each occurrence of R5aand R5bis independently selected from the group consisting of H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C8cycloalkyl, optionally substituted C2-C8heterocycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted C2-C8 heteroaryl; each occurrence of R6is ; each occurrence of R7aand R7b, ifselected from the group consisting of H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted C3-C8cycloalkyl, optionally substituted C2-C8heterocycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted C2-C8 heteroaryl; m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; each occurrence of n is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; - 168 - 53224792.3Attorney Docket No.0466483-7483WO1 o is 0, 1, 2, 3, 4, or 5; and each occurrence of p is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

40. The compound of claim 39, wherein at least one of the following applies: (a) R2a, R2b, and each occurrence of R2c, if present, are each independently H; (b) R3a, R3b, and each occurrence of R3c, if present, are each independently H; (c) each occurrence of R4aand R4bis independently H; (d) each occurrence of R5aand R5bis independently H; and (e) each occurrence of R7aand R7bis independently H.

41. The compound of claim 39 or 40, wherein the compound of formula (II) is selected from the group consisting of:

42. The compound of any one of claims 39-41, wherein R1a, R1b, and each occurrence of R1c, if present, are each independently , wherein: each occurrence of L1, ifselected from the group consisting of -(optionally substituted C1-C12alkylenyl)-, -(optionally substituted C2-C12alkenylenyl)-, - (optionally substituted C1-C12 alkynylenyl)-, -(optionally substituted C1-C12 heteroalkylenyl)- , -(optionally substituted C3-C8cycloalkylenyl)-, -(optionally substituted C2-C8heterocyloalkylenyl)-, -(optionally substituted C6-C10 arylenyl)-, -(optionally substituted C2- C8heteroarylenyl)-, -(optionally substituted C1-C12alkylenyl)-X-, -(optionally substituted C2- - 169 - 53224792.3Attorney Docket No.0466483-7483WO1 C12 alkenylenyl)-X-, -(optionally substituted C1-C12 alkynylenyl)-X-, -(optionally substituted C1-C12heteroalkylenyl)-X-, -(optionally substituted C3-C8cycloalkylenyl)-X-, -(optionally substituted C2-C8 heterocyloalkylenyl)-X-, -(optionally substituted C6-C10 arylenyl)-X-, - (optionally substituted C2-C8heteroarylenyl)-X-, and -X-; each occurrence of X, if present, is independently selected from the group consisting of -N(R8c)-, -N(RA)-, -O-, and -S-; each occurrence of R8a, R8b, and R8c, if present, is independently selected from the group consisting of optionally substituted C1-C20alkyl, optionally substituted C1-C20heteroalkyl, optionally substituted C2-C20 alkenyl, optionally substituted C2-C20 heteroalkyenyl, optionally substituted C2-C20alkynyl, and optionally substituted C2-C20heteroalkynyl, wherein one of R8aand R8bcan combine with one occurrence of L1to form an optionally substituted C2-C8 heterocycloalkyl; each occurrence of RAand RBis independently selected from the group consisting of H and optionally substituted C1-C6 alkyl; and each occurrence of q is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

43. The compound of claim 42, wherein each occurrence of L1is independently selected from the group consisting of -(CH2)1-5-, -(CH2)1-5CH(ORA)-, -(CH2)1-5O-, -(CH2)1-5N(RA)-, - .

44. The compound of claim 42 or 43, wherein -(L1)p- is selected from the group consisting of: ,- 170 - 53224792.3Attorney Docket No.0466483-7483WO1 ,45. The compound of any one of claims 39-44, wherein R1a, and each occurrence of R1c, if present, is independently selected from the group consisting of: , ,- 171 - 53224792.3Attorney Docket No.0466483-7483WO1 ,46. The compound of any one of claims 42-45, wherein each occurrence of R8a, R8b, and R8cis independently selected from the group consisting of -(CH2)CH(OH)(optionally substituted C3-C20alkyl).

47. The compound of any one of claims 42-46, wherein each R8a, R8b, and R8cis independently selected from the group consisting of -(CH2)CH(OH)(CH2)3CH3, - (CH2)CH(OH)(CH2)3CH3, -(CH2)CH(OH)(CH2)4CH3, -(CH2)CH(OH)(CH2)5CH3, - (CH2)CH(OH)(CH2)6CH3, -(CH2)CH(OH)(CH2)7CH3, -(CH2)CH(OH)(CH2)8CH3, - (CH2)CH(OH)(CH2)9CH3, -(CH2)CH(OH)(CH2)10CH3, -(CH2)CH(OH)(CH2)11CH3, - - 172 - 53224792.3Attorney Docket No.0466483-7483WO1 (CH2)CH(OH)(CH2)12CH3, -(CH2)CH(OH)(CH2)13CH3, -(CH2)CH(OH)(CH2)14CH3, - (CH2)CH(OH)(CH2)15CH3, -(CH2)CH(OH)(CH2)16CH3, -(CH2)CH(OH)(CH2)17CH3, - (CH2)CH(OH)(CH2)18CH3, -(CH2)CH(OH)(CH2)19CH3, and -(CH2)CH(OH)(CH2)20CH3.

48. The compound of any one of claims 39-47, wherein R1a, R1b, and each occurrence of R1c, if present, is independently selected from the group consisting of: , ,- 173 - 53224792.3Attorney Docket No.0466483-7483WO1 ,- 174 - 53224792.3Attorney Docket No.0466483-7483WO149. The compound of any one of claims 39-48, wherein the compound of formula (II) is:yl)propyl)adipimidamide, (C12-2aN).

50. A lipid nanoparticle (LNP) comprising: (a) at least one ionizable lipid comprising the compound of any one of claims 39- 49; (b) at least one neutral lipid; (c) at least one cholesterol lipid and / or a modified derivative thereof; and (d) at least one polymer-conjugated lipid and / or a modified derivative thereof.

51. The LNP of claim 50, wherein the at least one ionizable lipid compound comprises about 10 mol% to about 90 mol% of the LNP, optionally wherein the at least one ionizable lipid compound comprises about 57.1 mol% or about 50 mol% of the LNP.

52. The LNP of claim 50 or 51, wherein the at least ionizable lipid compound comprises or consists essentially of: - 175 - 53224792.3Attorney Docket No.0466483-7483WO1(C12-2aN).

53. The LNP of any one of claims 50-52, wherein the at least one neutral lipid comprises about 1 mol% to about 40 mol% of the LNP, optionally wherein the at least one neutral lipid comprises about 10.2 mol% or about 10.0 mol% of the LNP.

54. The LNP of any one of claims 50-53, wherein the neutral lipid comprises or consists essentially of at least one neutral lipid selected from the group consisting of dioleoylphosphatidylethanolamine (DOPE), distearoylphosphatidylcholine (DSPC), and dioleoylphosphatidylcholine (DOPC), optionally wherein the neutral lipid comprises or consists essentially of dioleoylphosphatidylethanolamine (DOPE).

55. The LNP of any one of claims 50-54, wherein the at least one cholesterol lipid and / or modified derivative thereof comprises about 20 mol% to about 75 mol% of the LNP, optionally wherein the at least one cholesterol lipid and / or modified derivative thereof comprises about 31.1 mol% or about 38.5 mol% of the LNP.

56. The LNP of any one of claims 50-55, wherein the at least one cholesterol lipid and / or modified derivative thereof comprises or consists essentially of cholesterol.

57. The LNP of any one of claims 50-56, wherein the at least one polymer-conjugated lipid comprises about 0.1 mol% to about 15 mol% of the LNP, optionally wherein the at least one polymer-conjugated lipid comprises about 1.7 mol% or about 1.5 mol% of the LNP.

58. The LNP of any one of claims 50-57, wherein the at least one polymer-conjugated lipid comprises or consists essentially of 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine- N-[methoxy(polyethyleneglycol)-2000] (C14PEG2K). - 176 - 53224792.3Attorney Docket No.0466483-7483WO1 59. The LNP of any one of claims 50-58, wherein the LNP has a molar ratio of (a) : (b) : (c) : (d) of about 57.1 : 10.2 : 31.1 : 1.7 or about 50 : 10 : 38.5 : 1.

5.

60. The LNP of any one of claims 50-59, wherein the LNP further comprises at least one cargo molecule, optionally wherein the at least one cargo molecule comprises or consists of a therapeutic cargo molecule.

61. The LNP of claim 60, wherein the cargo molecule is at least one selected from the group consisting of a nucleic acid, small molecule, protein, therapeutic agent, antibody, and any combinations thereof.

62. The LNP of claim 60 or 61, wherein the cargo molecule comprises a nucleic acid.

63. The LNP of claim 62, wherein the nucleic acid is DNA or RNA.

64. The LNP of claim 62 or 63, wherein the nucleic acid is selected from the group consisting of mRNA, cDNA, pDNA, microRNA, sgRNA, siRNA, modified RNA, antagomir, antisense molecule, and any combinations thereof.

65. The LNP of claim 64, wherein the mRNA encodes an enzyme, receptor, or antigen binding domain.

66. The LNP of claim 64, wherein the mRNA encodes a clustered regularly interspaced short palindrome repeats (CRISPR) associated protein, optionally wherein the CRISPR associated protein is Cas9.

67. The LNP of claim 66, wherein the nucleic acid cargo further comprises sgRNA.

68. A pharmaceutical composition comprising the lipid nanoparticle (LNP) of any one of claims 50-67 and at least one pharmaceutically acceptable carrier.

69. A method of treating, preventing, and / or ameliorating a disease in a subject, the method comprising administering to the subject at least one lipid nanoparticle (LNP) of any - 177 - 53224792.3Attorney Docket No.0466483-7483WO1 one of claims 50-67 or the pharmaceutical composition of claim 68.

70. The method of claim 69, wherein the disease is at least one selected from the group consisting of cancer, immune-mediated diseases, cardiovascular disease, and a metabolic disease.

71. A method of delivering at least one therapeutic cargo molecule to a dendritic cell of a subject, the method comprising administering to the subject at least one lipid nanoparticle (LNP) of any one of claims 50-67 or the pharmaceutical composition of claim 68.

72. The method of claim 71, wherein the therapeutic cargo molecule is mRNA.

73. The method of claim 72, wherein the LNP stimulates glycolysis in the dendritic cell.

74. The method of claim 72 or 73, wherein the LNP facilitates endosomal escape. - 178 - 53224792.3

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