Degradable ionizable lipid compounds, lipid nanoparticles (LNPS) comprising same, and methods of use thereof

Abstract

The present disclosure relates, in one aspect, to degradable ionizable lipid compounds of formula (I). In another aspect, the disclosure relates to lipid nanoparticles (LNPs) comprising at least one ionizable lipid of formula (I). In another aspect, the disclosure provides methods of the LNPs of the disclosure for splenic delivery of therapeutic cargo.

Description

[0001]Attorney Docket No.046483-7480WO1(04054) TITLE OF THE INVENTION Degradable Ionizable Lipid Compounds, Lipid Nanoparticles (LNPs) Comprising Same, and Methods of Use Thereof CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority under 35 U.S.C. § 119(e) to U.S. Provisional Patent Application No.63 / 708,052, filed October 16, 2024, which is incorporated herein by reference in its entirety. 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. BACKGROUND OF THE INVENTION 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. Recently, the U.S. Food and Drug Administration (FDA) approved two COVID-19 mRNA vaccines and one base editing therapy, all delivered by LNPs, to combat a global crisis and rare disease, respectively, representing a significant advance for mRNA therapeutics. Simultaneously, LNP-mediated CRISPR / Cas9-based gene therapies has showcased robust gene editing capabilities in clinical trials Within LNPs, ionizable lipids play critical roles in targeting specific tissues or cells, facilitating efficient translation, and providing on-demand adjuvant effects. Despite these advancements, 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. There is thus a need in the art for ionizable lipids and lipid nanoparticles comprising the same which permit targeting of extrahepatic tissues for enhanced mRNA therapeutic delivery. The present disclosure addresses this need. BRIEF SUMMARY OF THE INVENTION - 1 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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 and ; 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-C8 cycloalkylenyl)-, -(optionally substituted C2-C8 heterocyloalkylenyl)-, -(optionally substituted C6-C10 arylenyl)-, -(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-; R1a, R1b, R1c, R1d, and R1e, if present, is independently selected from the group consisting of and optionally substituted C1-C6 alkyl, wherein: more than two of R1a, R1b, R1c, R1d, and R1eare C1-C6alkyl, 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-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)-; - 2 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) each occurrence of R3aand R3bis independently selected from the group consisting of optionally substituted C1-C24alkyl and optionally substituted C1-C24heteroalkyl; each occurrence of R4is independently selected from the group consisting of H and optionally substituted C1-C6alkyl; 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 and optionally substituted C1-C6alkyl. In certain embodiments, the compound of formula (I) is: , In another aspect, the disclosure provides a lipid nanoparticle (LNP). In certain embodiments, the LNP comprises at least one ionizable lipid comprises at least one compound of the disclosure. In certain embodiments, the LNP comprises at least one 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 another aspect, the disclosure provides a pharmaceutical composition comprising the lipid nanoparticle (LNP) of the disclosure and at least one pharmaceutically acceptable carrier. In another aspect, the disclosure provide 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. In another aspect, the disclosure provides a method of delivering at least one therapeutic cargo molecule to a subject’s spleen. In certain embodiments, the method comprises administering to the subject at least one lipid nanoparticle (LNP) of the disclosure. - 3 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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-1C: Multiarm-assisted design of CODE LNP for mRNA delivery. FIG.1A: Comb-architecture polymer inspires design of comb-like degradable ionizable lipids. FIG. 1B: Chemical structures of 18 amine cores (left) and 15 multiarmed degradable alkyl tails (right) for generating 270 CODE ionizable lipids used in this study. FIG.1C: CODE LNPs were formulated via a microfluidic mixing device with CODE lipids, helper lipid, cholesterol, and PEG-lipid. The resulting CODE LNPs facilitate in vivo mRNA delivery to the spleen. FIGs.2A-2H: Structure-activity relationship (SAR) of CODE LNPs for FLuc mRNA delivery in vitro. FIG.2A: CODE LNP formulation parameters. CODE LNPs were formulated with one of 270 CODE lipids, DOPE, cholesterol, and C14PEG2K at a molar ratio of 35:16:46.5:2.5, respectively, for a total of 270 LNP formulations. FIG.2B: Representative cryogenic transmission electron microscopy (cryo-TEM) image of CODE LNP morphology. Scale bar: 50 nm. FIG.2C: Representative hydrodynamic size of CODE LNP revealed by DLS. FIG.2D: A heatmap of luciferase expression following treatment of HeLa cells with CODE LNPs (10 ng luciferase mRNA, n = 3 replicates). Relative luminescence unit (RLU) values of > 200 were calculated as hits for hit rate calculation. FIG. 2E: Relative hit rate of CODE LNPs with different secondary amine numbers. FIG.2F: Relative hit rate of CODE LNPs with different tail length. FIG.2G: Relative hit rate of CODE LNPs with different tail substitution number. FIG.2H: Relative hit rate of CODE LNPs with different tail architectures. FIGs.3A-3D: CODE LNPs-mediated in vivo delivery of mRNA to the spleen. FIG. 3A: In vivo evaluation of 14 CODE LNPs encapsulating FLuc mRNA (dose: 0.1 mg / kg). Representative bioluminescence IVIS images of main organs taken 12 h after systemic administration of CODE LNPs to C57BL / 6J mice. H: heart; Li: liver; S: spleen; Lu: lungs; K: kidneys. FIG.3B: Quantified luciferase mRNA expression in the spleen from FIG.3A. FIGs. 3C-3D: The spleen-targeting specificity was evaluated by calculating the relative luciferase expression of the Spleen / Liver (FIG.3C) and Spleen / Lungs (FIG.3D) (n = 3 mice). FIGs.4A-4D: 18-2-9b2 LNPs-mediated in vivo mRNA delivery of the splenic red pulp macrophage (RPM). FIG.4A: Ai14 mice were treated with 18-2-9b2 or MC3 / 18PA LNP encapsulating Cre mRNA 3 days prior to analysis (0.3 mg / kg). Spleen were digested and stained for quantifying cell populations for tdTomato+ expression. PBS was injected as negative control. FIG.4B: Physical characterizations of 18-2-9b2 and MC3 / 18PA LNPs. FIG. - 4 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 4C: Proportion of tdTomato+ cell types in the spleen assessed by flow cytometry. FIG.4D: Representative immunostaining demonstrating signal overlap between tdTomato+ cells and RPM marker CD163. DAPI was used for nuclear staining. F4 / 80 was used for macrophage staining. Statistical significance in FIG.4C was calculated using one-way analysis of variance (ANOVA), followed by Dunnett’s multiple comparison test. *P < 0.05; **P < 0.01. Data are presented as mean ± s.e.m. (n = 3 mice). FIGs.5A-5D: 18-2-9b2 LNPs-mediated in vivo BACH1 mRNA delivery of the spleen of Spic-GFP reporter mice. FIG.5A: 18-2-9b2 LNP-mediated BACH1 expression can repress the expression of Spic, leading to the downregulation of GFP signal in a Spic-GFP reporter mice model. LNP was systemically administered into the mice with a dosage of 1.0 mg / kg. FIG.5B: BACH1 level in sorted RPM was evaluated by quantitative PCR. FIG.5C: Proportion of GFP+ RPM assessed by flow cytometry. FIG.5D: Representative immunostaining demonstrating decreased GFP signal after BACH1 mRNA delivery. DAPI was used for nuclear staining. F4 / 80 was used for macrophage staining. CD163 was used for RPM staining. Statistical significance in FIGs.5B-5C was calculated using a Student’s t test with unpaired design. *P < 0.05. Data are presented as mean ± s.e.m. (n = 3 mice). FIG.6: Synthetic route of multiarmed acrylate-based tails. FIGs.7A-7O: Cell viability of HeLa cells after 24 h transfection of CODE LNPs derived from ionizable lipids prepared from amine heads 1-18 and A-D-5 (FIG.7A), A-D-6 (FIG.7B), A-D-6b (FIG.7C), A-D-7 (FIG.7D), A-D-7b (FIG.7E), A-D-7b2 (FIG.7F), A-D- 8 (FIG.7G), A-D-8b (FIG.7H), A-D-9 (FIG.7I), A-D-9b (FIG.7J), A-D-9b2 (FIG.7K), A- D-9b3 (FIG.7L), A-D-9v (FIG.7M), A-D-10 (FIG.7N), and A-D-11 (FIG.7O).5000 cells were plated per well and treated by 10 ng mRNA. Data are represented as mean ± s.e.m. (n = 3 replicates). FIG.8: Relative luminescence intensity of CODE LNPs in HeLa cells.5000 cells were plated per well and treated by 10 ng mRNA. MC3 LNP treated groups were used as a positive control. Medium treated groups were used as negative control. Data are represented as mean ± s.e.m. (n = 3 replicates). FIGs.9A-9D: In vivo Fluc luminescence intensity. FIG.9A: Physical characterization of 18-1-9b2 LNP. FIG.9B: Luminescence imaging of the body after LNP treatment. FIG.9C: Luminescence imaging of the organs from FIG.9B. H: heart; Li: Liver; S: spleen; Lu: lungs; K: kidneys. FIG.9D: Luminescence quantification of the organs from FIG.9C. Data are represented as mean ± s.e.m. (n = 2 mice). - 5 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) FIGs.10A-10B: TNS fluorescence curves for 18-1-9b2 (FIG.10A) and 18-2-9b2 (FIG.10B) LNPs. The apparent pKa of an LNP is computed as the pH at which 50% of the TNS fluorescence is measured. FIGs.11A-11D: Representative gating strategy of diverse tdTomato+cell types in the spleen. eFluor780 was used for distinguish live and dead cells. F4 / 80+was used to stain macrophages, then F4 / 80+ / CD163+was used for red pulp macrophages, F4 / 80- / CD3+was used for T cells, F4 / 80- / CD19+was used for B cells, and F4 / 80- / CD11c+was used for dendritic cells. Ai14 mice was administered with PBS or Cre mRNA LNP (MC3 / 18PA or 18- 2-9b2 LNP) at a total mRNA dosage of 0.3 mg / kg. The mice were necropsied 3 days post- injection for flow studies. (n = 3 mice). FIG.12: H&E staining of heart, liver, spleen, lungs, and kidney demonstrates minimal in vivo toxicity of 18-2-9b2 LNP compared with PBS treated groups. FIG.13: Representative gating strategy of GFP+red pulp macrophages in the spleen of Spic- GFP reporter mice. Draq7 was used for distinguish live and dead cells. F4 / 80+was used to stain macrophages, then F4 / 80+ / CD163+was used for red pulp macrophages. Spic-GFP reporter mice was administered with PBS or BACH1 mRNA 18-2-9b2 LNP at a total mRNA dosage of 1.0 mg / kg. The mice were necropsied 2 days post-injection for flow studies. (n = 3 mice). FIGs.14A-14H: Proteomics analysis of protein corona bounded on the LNP surface. FIG.14A: Schematic illustration of the experimental process of getting the protein corona adsorbed on the LNP for proteomics analysis. FIG.14B: Heatmap of different clusters in the protein corona between 18 and 2-9b2, MC3 / 18PA, and MC3-treated groups (n = 4). FIGs. 14C-14D: Pie chart (FIG.14C) and proportion (FIG.14D) of Cluster 2 in different LNP- treated groups. FIG.14E: Representative enhanced fold change of proteins in Clusters 2 between different LNP-treated groups. FIG.14F: Schematic illustration of endogenous protein adsorption on LNP-mediated intracellular processing of mRNA delivery.18-2-9b2 LNPs were pro-coated with Itga2b and used to treat RAW264.7 cell. FIGs.14G-14H: Luciferase expression was used to evaluate mRNA transfection (FIG.14G), while flow cytometry was used to evaluate intracellular uptake of Cy5-tagged mRNA-LNP (FIG.14H). Normalized luciferase expression is reported as the mean ± SEM (n = 3 biological replicates). Statistical significance in FIG.14D and FIGs.14G-14H was calculated using one-way analysis of variance (ANOVA), followed by Dunnett’s multiple comparison test. **P < 0.01; ***P < 0.001; ****P < 0.0001. - 6 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) FIGs.15A-15C: Counterpart LNP-mediated in vivo mRNA delivery. FIG.15A: In vivo evaluation of 14 counterpart LNPs encapsulating FLuc mRNA (dose: 0.1 mg / kg). Representative bioluminescence IVIS images of main organs taken 12 h after systemic administration of counterpart LNPs to C57BL / 6J mice. H: heart, Li: liver, S: spleen, Lu: lungs, K: kidneys. FIG.15B: Quantified luciferase mRNA expression in the liver from FIG. 15A. FIG.15C: The liver-targeting specificity was evaluated by calculating the average luciferase expression of the liver / spleen. FIG.16: Correlation between the particle size of LNPs with and without dendron-like design and their spleen-tropism. FIG.17: Principal components analysis (PCA) indicates protein adsorption difference of protein corona-LNP pellet. LNP1: MC3 / 18PA LNP; LNP2: MC3 LNP; LNP3: 18-2-9b2 LNP. FIG.18: Proteins that have been detected in all of the replicates from protein corona- LNP pellets. LNP1: MC3 / 18PA LNP; LNP2: MC3 LNP; LNP3: 18-2-9b2 LNP. FIG.19: Correlation analysis after data filtering and normalization between technical replicates. LNP1: MC3 / 18PA LNP; LNP2: MC3 LNP; LNP3: 18-2-9b2 LNP. FIG.20: The GO (Gene Ontology) gene functional enrichment analysis revealed the potential biological functions in which the upregulated markers in Cluster 2 from protein- LNP pellets may be involved. FIGs.21A-21B: Representative enhanced fold change of Apoe (FIG.21A) and Vtn (FIG.21B) in Clusters 1 between different LNP treated groups. FIG.22: Proportion of tdTomato+T cell, B cell, and DCs in the spleen assessed by flow cytometry. Data are presented as mean ± s.e.m. (n=3 mice). DETAILED DESCRIPTION OF THE INVENTION 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, - 7 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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 Lipid nanoparticle (LNP) have emerged as pivotal vehicles for messenger RNA (mRNA) delivery to hepatocytes upon systemic administration and to antigen-presenting cells following intramuscular injection. However, achieving systemic mRNA delivery to non- hepatocytes (e.g., the spleen) remains challenging without the incorporation of targeting ligands such as antibodies, peptides, or small molecules. The spleen is an immune cell-enriched tissue with a pivotal role in maintaining the integrity of the blood, supporting the immune system, and contributing to hematopoiesis. Various strategies have been explored to facilitate mRNA-LNP delivery to the spleen. One approach involves the incorporation of active targeting ligand, including proteins and peptides. However, a potential limitation is that active targeting often necessitates multi-step - 8 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) manufacturing for LNP formulation, leading to weak reproducibility and translatability. Another approach includes the incorporation of anionic molecules as a fifth component into standard four-component LNPs, such as 1,2-dioleoyl-sn-glycero-3-phosphate (18PA) and fatty acids, or as post-modification moieties within cationic polymer design. Nevertheless, the complex synthesis process of certain anionic lipids and the highly negatively charged potential of the resulting LNPs limits the clinical application of spleen selective mRNA- LNPs. Therefore, a simple and effective ionizable lipid with intrinsic spleen-tropism has utility to fully realize the potential of mRNA drug delivery to the spleen. In one aspect, the disclosure describes the design and synthesis of a series of comb- like degradable (CODE) ionizable lipids and the formulation of a library of LNPs, comprising CODE ionizable lipids, for systemic mRNA delivery to the spleen (FIG.1A). Initially, various acrylate-based tails were synthesized through a multiarm-assisted design and subsequently reacted with diverse amine cores to generate a library of 270 CODE ionizable lipids (FIG.1B). Following high-throughput in vitro screening in HeLa cells with firefly luciferase (FLuc)-encoding mRNA, several CODE LNPs exhibited superior protein expression compared to a gold standard LNP formulation. These top-performing CODE LNPs were further tested in vivo, demonstrating robust mRNA delivery to the spleen. Moreover, the lead CODE LNP, 18-2-9b2, encapsulating Cre mRNA demonstrated selective and effective genome modification of splenic red pulp macrophages (RPM), surpassing the performance of spleen-tropic MC3 / 18PA formulation. Additionally, 18-2-9b2 encapsulating BACH-1 mRNA exhibited robust BACH-1 expression in the RPM, resulting in the downregulation of Spic-GFP expression in a Spic- GFP transgene model. Thus, in one aspect, the disclosure demonstrates selective mRNA delivery to the spleen using comb-like architectural design of ionizable lipids. These results underscore the significance of structural evolution in developing novel ionizable lipids for organ-specific delivery of therapeutic cargoes for next-generation protein replacement therapy and gene editing applications. 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 - 9 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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, 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 - 10 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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 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)3 wherein 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. - 11 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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 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, C18alkyl chains, ether linkages between the head group and alkyl chains, and 0 to 3 double - 12 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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 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 - 13 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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, 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 - 14 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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, 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, - 15 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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- 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, - 16 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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. 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. - 17 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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 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, - 18 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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 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 - 19 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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 (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. - 20 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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 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 - 21 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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 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 - 22 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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 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. - 23 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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 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 - 24 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) “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. 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 - 25 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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. Ionizable Lipid Compounds In one aspect, the disclosure provides an ionizable lipid compound of formula (I), or a salt, stereoisomer, or isotopologue thereof: (I), wherein: A is selected from the group consisting and ; each occurrence of L1, if present, is independently selected from the group consisting of -(optionally substituted C1-C12 alkylenyl)-, -(optionally substituted 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-; R1a, R1b, R1c, R1d, and R1e, if present, is independently selected from the group - 26 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) consisting of and optionally substituted C1-C6alkyl, wherein: no more than one of R1aand R1bis C1-C6 alkyl, no more than one of R1c, R1d, and R1e, if present, is C1-C6 alkyl, 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 R3aand R3bis independently selected from the group consisting of optionally substituted C1-C24 alkyl and optionally substituted C1-C24 heteroalkyl; each occurrence of R4is independently selected from the group consisting of H and optionally substituted C1-C6 alkyl; 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 RAand RBare each independently selected from the group consisting of H and optionally substituted C1-C6 alkyl. 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(R1e)-. In certain embodiments, L1is . In certain embodiments, -(L1)m- is -(CH2)2-. In certain embodiments, -(L1)m- is - (CH2)3-. In certain embodiments, -(L1)m- is -(CH2)2N(CH3)(CH2)2-. In certain embodiments, - (L1)m- is -(CH2)3N(CH3)(CH2)3-. In certain embodiments, -(L1)m- is certain embodiments, -(L1)m- is - 27 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) is R2is C1-C6alkyl one group of ORA, N(RA)(RB), pyrrolidinyl, piperidinyl, and piperazinyl. In certain embodiments, R2is -(CH2)2N(CH3)2. In certain embodiments, R2is - (CH2)2N(CH2CH3)2. In certain embodiments, R2is -(CH2)3N(CH2CH3)2. In certain embodiments, R2is -(CH2)4OH. In certain embodiments, R2is . In certain . In certain embodiments, R2. In certain embodiments, R2. In certain . In certain of formula (I) (Ia): (Ia). In certain embodiments, the compound of formula (I) is a compound of formula (Ib). In certain embodiments, the . a : (Ie). In certain embodiments, the compound of formula (I) is a compound - 28 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) of formula (If) (If). In certain embodiments, the compound of formula (I) is a compound of formula (Ig): (Ig). In certain embodiments, the compound of formula (I) is a compound of formula (Ih): (Ih). In certain embodiments, the compound of formula (I) is a compound of formula (Ii): RARB1aN N R R1b(Ii). In certain embodiments, the compound of formula (I) is a of the compound of formula (I) is a compound of formula (Il). In certain embodiments, the compound of formula (I) is a : (Im). In certain embodiments, the compound of formula (I) is a compound of formula In certain embodiments, the : In certain embodiments, the compound of - 29 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) . RAis ethyl. In certain ethyl. In certain embodiments, L2is -(CH2)1-2-. In certain embodiments, L2is -O-. In certain embodiments, L2is -C(=O)-. In certain embodiments, -(L2)n- is -(CH2)2C(=O)O(CH2)-. In certain embodiments, R3ais -(CH2)OC(=O)(C5-C20alkyl). In certain embodiments, R3bis -(CH2)OC(=O)(C5-C20 alkyl). In certain embodiments, the C5-C20 alkyl is optionally substituted with at least one C1-C3alkyl. In certain embodiments, R1a, R1b, R1c, R1d, and R1e, if present, are each independently , wherein: each independently -C(=O)(optionally substituted C5-C20 alkyl); and each occurrence of o is independently 1, 2, 3, 4, or 5; and p, q, and r are each independently 1, 2, or 3. In certain embodiments, o is 2. In certain embodiments, p is 1. In certain embodiments, q is 1. In certain embodiments, r is 1. In certain embodiments, R5a. In certain embodiments, R5ais embodiments, R5a. In certain embodiments, . In certain embodiments, R5a. In certain embodiments, R5a. In certain embodiments, R5aIn certain embodiments, R5a. In certain embodiments, R5a. - 30 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) In certain embodiments, R5ais . In certain embodiments, R5ais embodiments, R5ais . In certain embodiments, . In certain embodiments, R5a. In certain O embodiments, R5ais . In certain embodiments, R5a. In certain embodiments, R5a. In certain embodiments, R5ais . In certain embodiments, R5a. In certain embodiments, R5a. In certain embodiments, R5a. In certain embodiments, R5a. In certain embodiments, R5ais . In certain embodiments, R5ais . In certain O embodiments, R5a. In certain embodiments, R5ais O . In certain embodiments, R5a. In certain embodiments, R5a. In certain embodiments, R5ais - 31 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) O . In certain embodiments, R5a. In certain O embodiments, R5a. In certain embodiments, R5ais O . In certain embodiments, R5a. In certain . In is . In certain embodiments, R5a. In certain embodiments, R5a. In certain embodiments, R5ais . In certain embodiments, R5a. In certain embodiments, R5ais . In certain embodiments, R5ais O O . In certain embodiments, R5a. In certain embodiments, R5a. In certain embodiments, R5ais O . In certain embodiments, R5a. In certain embodiments, R5a. In certain embodiments, R5ais In - 32 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) In In certain embodiments, R5a. In certain embodiments, R5bis . In certain embodiments, R5b. In certain embodiments, . In certain embodiments, R5b. In certain embodiments, R5bis embodiments, R5b. In certain embodiments, . In certain embodiments, R5b. In certain - 33 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) embodiments, R5b. In certain embodiments, R5bis . In certain embodiments, R5bis . In certain embodiments, R5bis . In certain embodiments, R5bis . In certain embodiments, R5bcertain embodiments, R5b. In certain embodiments, . In certain embodiments, R5bis . In certain embodiments, R5b. In certain embodiments, R5bis . In certain embodiments, R5bis O . In certain embodiments, R5bis . In certain O embodiments, R5b. In certain embodiments, R5bis Attorney Docket No.046483-7480WO1(04054) O embodiments, R5b. In certain embodiments, R5bis . R5bis . In certain embodiments, R5b. In certain embodiments, R5bis O . In certain embodiments, R5bis . In certain embodiments, R5b. In certain embodiments, R5bis O . In certain embodiments, R5b. In certain O embodiments, R5b. In certain embodiments, R5bis O . In certain embodiments, R5b. In certain embodiments, R5bis . In certain embodiments, R5bis In certain embodiments, R5b. In certain embodiments, R5bis In certain embodiments, R5b. In certain embodiments, R5bis - 35 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) In . . R4is ethyl. In certain In certain certain embodiments, R1ais is is is is Attorney Docket No.046483-7480WO1(04054) is is . In certain In certain embodiments, R1ais is - 37 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) is is is is is O O . Attorney Docket No.046483-7480WO1(04054) In certain In certain embodiments, R1bis is is is - 39 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) is R1cis is is . In certain In certain embodiments, R1cis is R1dis - 40 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) is is is is - 41 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) O O O O . In certain In certain embodiments, R1dis is is is - 42 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) is is is . In certain In certain embodiments, R1eis is R1eis - 43 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) . is selected from the group consisting of 1-2-5, 2-5, 9-2-5, 10-2-5, 11-2-5, 12-2-5, 13-2-5, 14-2-5, 15-2-5, 16-2-5, 17-2-5, 18-2-5, 1-2-6, 2-2-6, 3-2-6, 4-2-6, 5-2-6, 6-2-6, 7-2-6, 8-2-6, 9-2-6, 10-2-6, 11-2-6, 12-2-6, 13-2-6, 14-2-6, 15-2-6, 16-2-6, 17-2-6, 18-2-6, 1-2-6b, 2-2-6b, 3-2- 6b, 4-2-6b, 5-2-6b, 6-2-6b, 7-2-6b, 8-2-6b, 9-2-6b, 10-2-6b, 11-2-6b, 12-2-6b, 13-2-6b, 14-2- 6b, 15-2-6b, 16-2-6b, 17-2-6b, 18-2-6b, 1-2-7, 2-2-7, 3-2-7, 4-2-7, 5-2-7, 6-2-7, 7-2-7, 8-2-7, 9-2-7, 10-2-7, 11-2-7, 12-2-7, 13-2-7, 14-2-7, 15-2-7, 16-2-7, 17-2-7, 18-2-7, 1-2-7b, 2-2-7b, 3-2-7b, 4-2-7b, 5-2-7b, 6-2-7b, 7-2-7b, 8-2-7b, 9-2-7b, 10-2-7b, 11-2-7b, 12-2-7b, 13-2-7b, 14-2-7b, 15-2-7b, 16-2-7b, 17-2-7b, 18-2-7b, 1-2-7b2, 2-2-7b2, 3-2-7b2, 4-2-7b2, 5-2-7b2, 6- 2-7b2, 7-2-7b2, 8-2-7b2, 9-2-7b2, 10-2-7b2, 11-2-7b2, 12-2-7b2, 13-2-7b2, 14-2-7b2, 15-2- 7b2, 16-2-7b2, 17-2-7b2, 18-2-7b2, 1-2-8, 2-2-8, 3-2-8, 4-2-8, 5-2-8, 6-2-8, 7-2-8, 8-2-8, 9-2- 8, 10-2-8, 11-2-8, 12-2-8, 13-2-8, 14-2-8, 15-2-8, 16-2-8, 17-2-8, 18-2-8, 1-2-8b, 2-2-8b, 3-2- 8b, 4-2-8b, 5-2-8b, 6-2-8b, 7-2-8b, 8-2-8b, 9-2-8b, 10-2-8b, 11-2-8b, 12-2-8b, 13-2-8b, 14-2- 8b, 15-2-8b, 16-2-8b, 17-2-8b, 18-2-8b, 1-2-9, 2-2-9, 3-2-9, 4-2-9, 5-2-9, 6-2-9, 7-2-9, 8-2-9, 9-2-9, 10-2-9, 11-2-9, 12-2-9, 13-2-9, 14-2-9, 15-2-9, 16-2-9, 17-2-9, 18-2-9, 1-2-9b, 2-2-9b, 3-2-9b, 4-2-9b, 5-2-9b, 6-2-9b, 7-2-9b, 8-2-9b, 9-2-9b, 10-2-9b, 11-2-9b, 12-2-9b, 13-2-9b, 14-2-9b, 15-2-9b, 16-2-9b, 17-2-9b, 18-2-9b, 1-2-9b2, 2-2-9b2, 3-2-9b2, 4-2-9b2, 5-2-9b2, 6- 2-9b2, 7-2-9b2, 8-2-9b2, 9-2-9b2, 10-2-9b2, 11-2-9b2, 12-2-9b2, 13-2-9b2, 14-2-9b2, 15-2- 9b2, 16-2-9b2, 17-2-9b2, 18-2-9b2, 1-2-9b3, 2-2-9b3, 3-2-9b3, 4-2-9b3, 5-2-9b3, 6-2-9b3, 7- 2-9b3, 8-2-9b3, 9-2-9b3, 10-2-9b3, 11-2-9b3, 12-2-9b3, 13-2-9b3, 14-2-9b3, 15-2-9b3, 16-2- 9b3, 17-2-9b3, 18-2-9b3, 1-2-9v, 2-2-9v, 3-2-9v, 4-2-9v, 5-2-9v, 6-2-9v, 7-2-9v, 8-2-9v, 9-2- 9v, 10-2-9v, 11-2-9v, 12-2-9v, 13-2-9v, 14-2-9v, 15-2-9v, 16-2-9v, 17-2-9v, 18-2-9v, 1-2-10, 2-2-10, 3-2-10, 4-2-10, 5-2-10, 6-2-10, 7-2-10, 8-2-10, 9-2-10, 10-2-10, 11-2-10, 12-2-10, 13-2-10, 14-2-10, 15-2-10, 16-2-10, 17-2-10, 18-2-10, 1-2-11, 2-2-11, 3-2-11, 4-2-11, 5-2-11, 6-2-11, 7-2-11, 8-2-11, 9-2-11, 10-2-11, 11-2-11, 12-2-11, 13-2-11, 14-2-11, 15-2-11, 16-2- 11, 17-2-11, and 18-2-11. Exemplary nomenclature includes “X-#-Y, wherein “X” is an integer between 1 and 18 which indicates the identity of the amine core, “#” represents the number of multiarmed degradable tails (e.g., when “#” is 2, there are chains in each tail), and “Y” is an integer which represents the carbon on each tail. Additionally, “b” used in conjunction with - 44 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) “Y” herein indicates branching of the alkyl chain thereof (e.g., “b,” “b2,” and “b3,” indicating substitution with a C1, C2, or C3alkyl, respectively, which may be substituted at the 1- or 3- position of the acyl moiety). Further, “v” used in conjunction with the “Y” term refers to a point of unsaturation (i.e., presence of a double bond); in non-limiting embodiments, the point of unsaturation may be 2,3-unsaturation of the acyl moiety. Ionizable lipids exemplified herein represent # is 2. In certain embodiments, the compound of formula (I) is: , 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-C8heterocycloalkyl (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 - 45 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) limited to ionizable lipidoids of formula (I). 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- 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- - 46 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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, 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, - 47 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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 can also be used. The acyl groups in these lipids can be, for example, acyl groups derived from fatty acids having C10-C24 carbon 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 - 48 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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. 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 - 49 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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% 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) 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 - 50 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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 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 ionizable lipid compound comprises: . of: - 51 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) . 1, 2, 3, 4, 5, 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, 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 - 52 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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, 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, - 53 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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 comprises BACH1 mRNA. In certain embodiments, the mRNA encodes an enzyme. 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. - 54 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) Methods 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. In certain embodiments, the disease is at least one selected from the group consisting of cancer, cardiovascular disease, and a metabolic disease. In certain embodiments, the disease relates to aberrant red blood cell differentiation. In certain embodiments, the disease is associated with BACH1 overexpression. In another aspect, the disclosure provides a method of delivering at least one therapeutic cargo molecule to a subject’s spleen. In certain embodiments, the method comprises administering to the subject at least one lipid nanoparticle (LNP) of the disclosure. 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 - 55 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) mRNA, cDNA, pDNA, microRNA, sgRNA, siRNA, modified RNA, antagomir, antisense molecule, and any combinations thereof. In certain embodiments, the mRNA comprises BACH1 mRNA. In certain embodiments, the mRNA encodes an enzyme. In certain embodiments, the mRNA encodes a clustered regularly interspaced short palindrome repeats (CRISPR) associated protein, optionally wherein the CRISPR associated protein is Cas9. In certain embodiments, the nucleic acid cargo further comprises sgRNA. 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. 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 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 - 56 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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. 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 - 57 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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., 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, - 58 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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, 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, - 59 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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 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 - 60 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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 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 - 61 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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 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 - 62 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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 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 - 63 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) (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 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 - 64 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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. 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 - 65 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 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. Materials and Methods Formulation of CODE ionizable lipids into lipid nanoparticles (LNPs) All LNPs used in this study were prepared as follows. An ethanol phase containing all lipids and an aqueous phase containing mRNA (FLuc mRNA, Cre mRNA or BACH1 mRNA) were mixed using a microfluidic device to formulate LNPs. The ethanol phase contained CODE 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. Aqueous phase was composed of 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. It was noted that MC3 LNP formulation was formulated with ethanol phase containing MC3 lipid, DSPC, cholesterol, and C14PEG2K with a fixed molar ratio of 50%, 10%, 38.5%, and 1.5%, respectively. MC3 / 18PA LNP was formulated with ethanol phase containing MC3 lipid, 18PA, DSPC, cholesterol, and C14PEG2K with a fixed molar ratio of 35%, 30%, 7%, 27%, and 1.1%, respectively. Zetasizer Nano was used to measure the Z-average diameters, polydispersity index (PDI) and Zeta potential. mRNA concentration and encapsulation efficiency in each LNP formulation were measured using a modified Quant-iT RiboGreen (ThermoFisher) assay on a plate reader. In vitro FLuc mRNA LNP library screening In a white transparent 96-well plate, HeLa cells were seeded at a density of 5 x 103cells per well in 100 μL growth medium (DMEM, 10% FBS, 1% P / S), and were incubated at 37oC in 5% CO2. The medium was exchanged for fresh growth medium, and then LNPs were treated at a dose of 10 ng Luc mRNA per well. Firefly luciferase expression was measured 24 h after LNP transfection using a Luciferase Assay System (Promega) according to the manufacturer’s protocol. The luminescent signal was normalized to PBS treated cells. Cell viability was measured using a CellTiter-Glo Luminescent Cell Viability Assay (Promega), in which the luminescence was normalized to PBS treated cells according to the manufacturer’s protocol. - 66 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) In vivo FLuc mRNA LNP delivery All animal procedures were performed on female C57BL / 6J mice aged 6-8 weeks. Mice were administered a single intravenous FLuc mRNA at a dosage of 0.1 mg / kg via tail vein injection. The luciferase expression was evaluated using an IVIS Spectrum imaging system (Caliper Life Sciences) 12 h post-injection. Mice were then injected D-luciferin (PerkinElmer) at a dose of 150 mg / kg by intraperitoneal injection (IP). After 10 min incubation under anesthesia, bioluminescence intensity was quantified by measuring photon flux in the region of interest where signal emanated using Living IMAGE Software provided by Caliper. Ex vivo imaging was performed on heart, liver, spleen, lung, and kidney after resection. LNP-mediated Cre mRNA delivery to Ai14 mice B6.Cg-Gt(ROSA)26Sortm14(CAG-tdTomato)Hze / J (Ai14) Mice were administered a single intravenous Cre mRNA LNPs at a dosage of 0.3 mg / kg via tail vein injection. After 3 days, mice were euthanized and the splenic single cell suspensions were collected for flow cytometry measurement. The spleen was grinded into small pieces to release spleen cells. The obtained cell suspension was then centrifuged (5 min, 500 g) and lysed by ACK lysis buffer (ThermoFisher) (1 mL) for 10 min. Afterwards, single-cell suspensions were obtained by centrifugation (5 min, 500 g) and resuspended in 1x PBS (400 μL). The antibodies used were: anti-mouse BV421 F4 / 80 antibody (1:200, Biolegend, Cat#123137), APC CD163 antibody (1:200, Biolegend, Cat#155306), AF488 CD19 antibody (1:200, Biolegend, Cat#115521), BV711 CD11c antibody (1:200, Biolegend, Cat#117349). The obtained single-cell suspensions were stained at 4oC for 30 min by each of the above antibodies (2 μL), and afterwards were centrifuged, washed, centrifuged and resuspended in Live / dead staining eFluor7801x PBS (0.1%, 500 μL) for flow cytometry analysis. LNP-mediated BACH1 mRNA delivery to Spic-GFP reporter mice Spic-GFP report mice were administered a single intravenous injection of 18-2-9b2 LNPs encapsulating mRNA encoding for BACH1 at dosages ranging from 1.0 mg / kg via tail vein injection. After 48 h, mice were sacrificed and the spleen was collected. GFP expression of splenic red pulp macrophages were evaluated through flow cytometry. Splenic cell suspensions were obtained by a similar protocol described above. Anti-mouse BV421 F4 / 80 - 67 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) antibody (1:200, Biolegend, Cat#123137) and APC CD163 antibody (1:200, Biolegend, Cat#155306) were used for staining. Drap7 (0.1% in 1x PBS) was used for live / dead staining. Splenic red pulp macrophages were gated as F4 / 80+ / CD163+, and sorted for qPCR measurement. qPCR was conducted according to a previous study.3Total RNA was extracted by ReliaPrep RNA Cell Miniprep kit according to the manufacturer’s recommendation (Promega, Cat#Z6011) and then reverse-transcribed into complementary DNA (cDNA) using the High- Capacity RNA-to-cDNA kit (ThermoFisher Scientific, Cat#4387406). Afterwards, cDNA was dilute 4 times through adding 80 uL of water into 20uL of cDNA. Mastermix was made by a TaqMan Universal MasterMix and TaqMan probes. Then, 3.75 uL of mastermix and 1.25 uL of cDNA was added per well into a 384-well plate, and qPCR was performed on an Applied Biosystems QuantStudio 6 Real-Time PCR System (ThermoFisher Scientific). Immunofluorescence Tissue sections were obtained with the help from the Wistar Core (UPenn). Afterward, tissue sections were blocked in PBS + 3% BSA for 5 min at room temperature. For the samples from Spic-GFP reporter mice, slides were washed twice by 1x PBS and incubated with primary antibodies (F4 / 80, 1:200, Cell Signaling Technology, Cat#30325; CD163, 1:100, ThermoFisher Scientific, Cat#14-1631-82; GFP antibody, 1:1000, Rockland, Cat#600- 101-215) overnight at 4 °C. After, slides were washed and incubated with fluorophore- conjugated secondary antibodies (AF488-conjugated donkey anti-goat antibody, 1:1000, ThermoFisher Scientific, Cat#A-11055; AF568-conjugated donkey anti-rat antibody, 1:1000, ThermoFisher Scientific, Cat#A78946; AF647-conjugated donkey anti-rabbit antibody, 1:1000, ThermoFisher Scientific, Cat#A-31573) for 2 h. At last, slides were washed and incubated with 1 uM DAPI for 5 min, and mounted using ProLong Gold (Life Sciences, Cat#P36930). Standard multiplex immunofluorescent images were taken with a Leica Dmi8 microscope and analyzed with LAS X software (Leica). Samples from Ai14 mice were processed by a similar protocol described above. Primary antibodies include F4 / 80 (1:200, Cell Signaling Technology, Cat#30325) and CD163 (1:100, ThermoFisher Scientific, Cat#14-1631-82). Secondary antibodies include AF488- conjugated donkey anti-rat antibody, 1:1000, ThermoFisher Scientific, Cat#A-21208; AF647- conjugated donkey anti-rabbit antibody, 1:1000, ThermoFisher Scientific, Cat#A-31573). Example 1: Chemical Synthesis Synthesis of 2,2-bis(hydroxymethyl)butyl acrylate - 68 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 2-Ethyl-2- g, 0.15 mol, 5.0 equiv) and triethylamine (TEA, 4.18 mL, dissolved in anhydrous acetone (50 mL) and the mixture was cooled to 0 °C on an ice bath. Acryloyl chloride (2.71 g, 0.03 mol, 1.0 equiv) was added dropwise under vigorous stirring. The reaction was then allowed to warm to room temperature overnight. Afterwards, solvent was removed to obtain the crude product. The final pure compound was further purified by flash chromatographic (silica gel, gradient eluant from hexane / EA = 10 / 1 to hexane / EA = 2 / 1) as a colorless oil.1H NMR (400 MHz, CDCl3), δ 6.50-6.41 (d, 1H), 6.22-6.12 (m, 1H), 5.94-5.87 (d, 1H), 4.32 (s, 2H), 3.61 (s, 4H), 1.41-1.30 (m, 2H), 0.97-0.88 (m, 3H). LC-MS (m / z): Calcd for [M+H]+: 189.2, Found: 189.2. Exemplary synthesis of multiarmed acrylate-based tail A-D-5 An exemplary (acryloyloxy)methyl)-2- ethylpropane-1,3-diyl dihexanoate) is provided herein to demonstrate the general procedure utilized to prepare the multiarmed acrylate-based tail compounds described herein. Briefly, a flame-dried 50 mL round-bottom flask was charged with 2,2- bis(hydroxymethyl)butyl acrylate (188.22 mg, 1 mmol, 1.0 equiv), EDC^HCl (575.1 mg, 3 mmol, 3.0 equiv), DMAP (36.65 mg, 0.3 mmol, 0.3 equiv), hexanoic acid (348.48 mg, 3 mmol, 3.0 equiv) and DCM. The reaction was conducted overnight. Then, the solution was concentrated in vacuo and rinsed with saturated NaCl. The final product was afforded by flash chromatographic (silica gel, gradient eluant from hexane to hexane / DCM = 1 / 2) as a colorless oil.1H NMR (400 MHz, CDCl3), δ 6.45-6.38 (d, 1H), 6.19-6.07 (m, 1H), 5.90-5.85 (d, 1H), 4.15 (s, 2H), 4.05 (s, 4H), 2.41-2.28 (m, 4H), 1.74-1.49 (m, 6H), 1.41-1.24 (m, 8H), 0.99-0.87 (m, 9H). LC- MS (m / z): Calcd for [M+NH4]+: 402.5, Found: 402.4. Table 1. Exemplary multiarmed acrylate-based tails and characterization data thereof - 69 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) Name Compound Characterization Data O O1H NMR (400 MHz,CDCl3), δ 6.46-6.37d 1H 617608 1H 590584 d , - 70 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 486.7. O O1H NMR (400 MHz, CDCl3), δ 6.46- 639 (d 1H) 619-608 (m 1H) 591- - 71 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) O O1H NMR (400 MHz, CDCl3), δ 6.47- O O 6.39 (d, 1H), 6.19-6.08 (m, 1H), 5.91- 584 (d 1H) 415 (s 2H) 407 (s 4H) the general procedure utilized to prepare the multiarmed degradable ionizable lipids described herein. Core No.18 (50.7 mg, 0.167 mmol, 1 equiv) and A-D-9b2 (498 mg, 1 mmol, 6 equiv) were added in a glass vial equipped with a stir bar dissolved in ethanol (100 μL). The reaction was processed at 80 °C for 2 days. Then the solvent was removed and the resulting crude product was subjected for CombiFlash chromatography (DCM to DCM / Methanol = 20 / 1) to obtain a purified 18-2-9b2 ionizable lipids as a yellowish oil. CODE ionizable lipids can be used for LNP preparation without further purification.1H NMR (400 MHz, CDCl3), δ 4.14-3.99 (m, 30H), 3.61-3.42 (m, 10H), 2.95-2.78 (m, 16H), 2.71-2.62 (m, 12H), 2.52-2.42 (m, 10H), 2.36-2.25 (m, 20H), 1.64-1.54 (m, 20H), 1.53-1.48 (m, 10H), 1.37-1.20 (m, 90H), 0.96-0.82 (m, 75H).13C NMR (101 MHz, CDCl3), δ 173.7, 172.1, 63.6, 60.3, 53.0, 49.6, 40.6, 38.3, 32.4, 31.6, 28.8, 28.2, 25.5, 23.1, 22.9, 14.2, 14.1, 10.7, 7.4. LC-MS (m / z): Calcd for [M+2H]+: 1394.1, Found: 1394.2. In certain embodiments, alternative multiarmed degradable ionizable lipid were prepared according to the methods described for 18-2-9b2 utilizing alternative amine core species (Table 2). Table 2. Exemplary amine core species - 72 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) No No Structure Structure . . Synthesis of comparator (18-1-9b2) described herein for the preparation of 18-2-9b2, except that A-D-9b2 was substituted for 2- (acryloyloxy)ethyl 4-ethyloctanoate. The resultant product was purified by silica gel - 73 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) chromatograph to provide.18-1-9b2.1H NMR (400 MHz, CDCl3), δ 4.30-4.25 (m, 20H), 3.52-3.46 (m, 8H), 2.94-2.81 (m, 12H), 2.72-2.63 (m, 8H), 2.52-2.46 (m, 10H), 2.37-2.28 (m, 10H), 1.64-1.57 (m, 10H), 1.34-1.21 (m, 52H), 0.93-0.82 (m, 30H). LC-MS (m / z): Calcd for [M+2H]+: 828.2, Found: 828.2. Example 2: Multiarm-assisted design of comb-like degradable lipids Comb-like polymers comprising densely grafted chains and tunable attributes, such as composition, shape, stiffness, and surface properties, have emerged as a promisingplatform for biomedical research, drug delivery, and diagnostic applications. Notably,comb-architecture polymers have been increasingly utilized as effective vectors for RNA interference therapy, vaccination, and cancer therapy. Initially, a series of multiarmed acrylate-based tails were synthesized through a two-step nucleophilic acyl substitution. The reaction can occur in mild conditions with high yields. Then, these multiarmed acrylate- based tails were reacted with various amine cores through a Michael addition reaction, yielding a library of 270 comb-like degradable ionizable lipids. The resulting CODE lipid library with varying amine core structures, tail architectures, and tail substitution numbers, wherein CODE lipids are descriptively named, as described elsewhere herein. Exemplary nomenclature includes “X-#-Y, wherein “X” indicates the identity of the amine core, “#” represents the number of multiarmed degradable tails (e.g., when “#” is 2, there are 2 chains in each tail), and “Y” represents the carbon number on each tail. Additionally, “b” used in conjunction with “Y” herein indicates branching of the alkyl chain thereof (e.g., “b,” “b2,” and “b3,” indicating substitution with a C1, C2, or C3alkyl, respectively, which may be substituted at the 1- or 3-position of the acyl moiety). Further, “v” used in conjunction with the “Y” term refers to a point of unsaturation (i.e., presence of a double bond); in non-limiting embodiments, the point of unsaturation may be 2,3-unsaturation of the acyl moiety. Ionizable lipids exemplified herein represent # is 2. For example, ionizable lipid 18-2-9b2 refers to an ionizable lipid prepared by Michael addition (i.e., [1,4]-conjugate addition) of amine core 18 and α,β-unsaturated ester (i.e., “multiarmed degradable tail”) A-D-9b2, wherein both terminal primary amines thereof undergo two iterative Michael addition reaction, and the internal secondary thereof undergoes a single Michael addition reaction (see FIG.1B and Example 1). Additionally, the exemplary α,β-unsaturated esters referenced in the context of the full ionizable lipid as “2-9b2” and “1-9b2” differ not only with respect to the number of degradable tails, but also the linking moiety between the α,β-unsaturated ester moiety itself and the degradable tail. For - 74 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) example, the exemplary degradable tail referred to as “A-D-9b2”—or “2-9b2” in conjunction with the ionizable lipid—comprises an ethyl-substituted C4trivalent linker covalently linking the two degradable tails to the α,β-unsaturated ester moiety: . In contrast, the comprises an unsubstituted C2 divalent linker covalently unsaturated ester moiety. . Likewise, degradable the unsubstituted C2 divalent linker, whereas degradable tails referred to as “2-” comprise the ethyl-substituted C4trivalent linker. These CODE lipids can be mixed with phospholipid, cholesterol, lipid-anchored polyethylene glycol, and mRNA using a microfluidic mixing device to generate LNPs with distinct physiochemical properties (FIG.1C). This combinatorial CODE ionizable lipid library extends the chemical diversity of ionizable lipids for nucleic acid delivery applications. Example 3: Structure-activity relationship (SAR) of CODE LNP for FLuc mRNA delivery in vitro The structure-activity relationship (SAR) of CODE ionizable lipids for mRNA delivery was subsequently explored in vitro. CODE LNPs encapsulating FLuc mRNA were used to transfect HeLa cells, a cell line shown to be conducive to high-throughput in vitro screening of LNP formulations in previous studies. CODE LNPs were formulated by mixing an ethanol phase containing CODE lipids, DOPE, cholesterol, and C14PEG2K (35:16:46.5:2.5, molar ratio) and an aqueous phase containing FLuc mRNA via a microfluidic mixing approach (FIG.2A). CODE LNPs were characterized by particle size, polydispersity index (PDI), zeta potential, and mRNA encapsulation efficiency. The hydrodynamic diameter for all CODE LNP formulations ranged from 70 to 150 nm, as determined by intensity measurements using dynamic light scattering (DLS) (Table 3). - 75 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) Cryo-transmission electron microscopy (cryo-TEM) revealed a uniform solid core morphology in a representative CODE LNP (FIGs.2B-2C). The majority of CODE LNP formulations (>80%) exhibited high monodispersity, with a PDI value less than 0.2 (Table 3). Additionally, CODE LNPs displayed a relatively negative surface zeta potential and high mRNA encapsulation efficiency (Table 3). Table 3. Selected characterization data for exemplary CODE LNPs of the disclosure LNP name Diameter (nm) PDI Zeta potential (mV) EE (%) 1-2-5 85.9 ± 2.6 0.07 ± 0.05 -8.7 ± 1.5 75.6 - 76 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 2-2-6b 80.5 ± 3.7 0.15 ± 0.06 -3.6 ± 4.6 73.8 3-2-6b 88.9 ± 5.3 0.20 ± 0.08 -2.3 ± 2.2 82.4 426b 911 ± 45 015 ± 002 72 ± 45 801 - 77 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 14-2-7b 123.8 ± 7.4 0.21 ± 0.02 -9.4 ± 5.8 74.5 15-2-7b 117.0 ± 7.6 0.07 ± 0.01 -7.1 ± 4.3 78.8 1627b 1324 ± 37 010 ± 002 98 ± 64 809 - 78 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 8-2-8b 123.6 ± 9.8 0.22 ± 0.04 -6.1 ± 5.4 81.6 9-2-8b 133.0 ± 4.5 0.14 ± 0.02 -5.7 ± 5.1 84.5 1028b 1342 ± 47 006 ± 003 43 ± 25 783 - 79 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 2-2-9b2 75.6 ± 3.1 0.17 ± 0.05 4.2 ± 1.5 77.8 3-2-9b2 82.2 ± 4.3 0.12 ± 0.01 6.2 ± 0.5 72.6 429b2 1039 ± 64 005 ± 003 71 ± 09 757 - 80 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 14-2-9v 131.8 ± 5.6 0.21 ± 0.05 3.8 ± 2.2 70.6 15-2-9v 114.9 ± 7.6 0.22 ± 0.04 -7.5 ± 3.5 75.4 1629 1198 ± 18 016 ± 008 82 ± 67 768 Table 4. Selected characterization data for exemplary LNPs comprising ionizable lipids with only one alkyl chain in each tail group ) - 81 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 17-1-6 106.6 ± 5.3 0.17 ± 0.03 -22.6 ± 1.4 85.5 17-1-7b2 186.6 ± 2.7 0.11 ± 0.01 -17.0 ± 5.4 67.8 1719 1359 ± 51 014 ± 002 236 ± 18 739 CODE lipids-mediated mRNA delivery efficacy was generated (FIG.2D). To elucidate how structural parameters influenced mRNA delivery, the relative hit rate, defined as the value of relative luminescence unit (RLU) greater than 200, was assessed. The amine number of each CODE lipid that influenced mRNA delivery efficacy was first investigated. The data indicate that CODE lipids with five secondary amines per lipid exhibited the highest mRNA transfection with a hit rate of 12.6% across the entire library (FIG.2E). Notably, a higher number of secondary amine heads (>2) have been shown previously to facilitate endosomal escape of mRNA, leading to enhanced delivery efficacy. Additionally, tail length and tail substitution numbers have been shown to significantly influence mRNA delivery. Among CODE lipids, a tail length of 9 and tail substitution number of 10 resulted in the highest relative hit rate (FIGs.2F-2G). Moreover, CODE lipids with branched architectures demonstrated superior mRNA delivery compared to linear and unsaturated counterparts (FIG.2H), aligning with prior studies indicating that branched tails may enhance stability and fusogenicity for mRNA delivery. All CODE LNPs exhibited minimal toxicity in vitro (FIGs.7A-7O). Interestingly, 14 CODE LNP formulations in this library outperformed the gold standard MC3 LNP in vitro (FIG.2D and FIG.8). Thus, these 14 CODE LNP formulations were selected for subsequent investigation in vivo. Example 4: CODE LNPs facilitate in vivo mRNA delivery to the spleen 14 CODE LNPs were assessed for in vivo mRNA biodistribution in C57BL / 6J mice following systemic administration of 0.1 mg / kg FLuc mRNA. After 12 h, mice were scarified, and organs (i.e., heart, liver, spleen, lungs, and kidneys) were isolated to quantify luciferase expression using an in vivo imaging system (IVIS) (FIGs.3A-3B). Notably, these CODE LNPs exhibited variable mRNA delivery to the spleen. To assess spleen - 82 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) targeting specificity, the relative luciferase expression of Spleen / Liver and Spleen / Lungs of the above CODE LNPs was calculated based on the IVIS data (FIGs.3C-3D). Among these, 18-2-9b2, composed of five secondary amines and ten multiarmed degradable tails with a branched architecture, demonstrated the most efficient spleen-specific mRNA delivery. Although 15-2-9b1 and 18-2-11 exhibited greater luminescence intensity in the spleen than 18-2-9b2 LNP, they also showed higher transfection in the liver and lungs, indicating lower specificity for the spleen. Following the analysis of both spleen delivery efficacy and specificity, 18-2-9b2 LNP was selected for further consideration. To elucidate whether the comb-like design facilitates mRNA delivery to the spleen, a similar ionizable lipid without the comb-like structure, 18-1-9b2, was designed and synthesized. In vivo, 18-1-9b2 LNP mediated robust mRNA delivery to the liver, rather thanthe spleen (FIGs. 9A-9D). The pKavalues of 18-1-9b2 and 18-2-9b2 LNPs were theninvestigated (FIGs. 10A-10B). The pKavalue of 18-1-9b2was found to be 6.20,consistent with previous studies indicating pKa values between 6 and 7 for potent nucleic acid delivery to the liver (FIG.11A). Conversely, the pKa of 18-2-9b2 was 5.65, deviating from the established pKa characteristics associated with 18PA-based SORT spleen LNP (pka ranging from 4 to 5) for mRNA delivery to the spleen (FIG.11B). Therefore, the mechanism of spleen targeting of 18-2-9b2 LNPs was explored by assessing the protein corona ex vivo via mass spectrometry-based proteomics. Albumin, the most abundant serum protein, predominantly adsorbed onto 18-2-9b2 with an abundance above 15%. Interestingly, apolipoprotein E (ApoE, <0.2%) and vitronectin (Vtn, <0.1%) were not among the most abundant proteins within the corona, both of which have already been shown in previous studies to facilitate nucleic acid delivery to the liver and lungs, respectively, through specific receptor-mediated endocytosis. Furthermore, hemoglobin alpha 2 (HBA2) was significantly enriched (~10%) on the surface of 18-2-9b2 LNPs, which may facilitate red blood cell trafficking within the spleen, potentially directing LNP spleen-tropism. Thus, it was hypothesized that a shift away from LNP binding of ApoE and Vtn towards hemoglobin- related proteins facilitates spleen tropism of 18-2-9b2 LNP. The endogenous targeting potential of 18-2-9b2 LNPs was explored by assessing the protein corona ex vivo via mass spectrometry-based proteomics (FIG.14A and FIGs.17-19). There is a significant enhancement of Cluster 2 in the corona bound to the 18-2-9b2 LNP (57.3%), which is 3.23-fold and 2.11-fold enhancement compared to MC3 and MC3 / 18PA groups, respectively (FIGs.14B-14D). A further Gene Ontology (GO) enrichment analysis of - 83 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) Cluster 2 revealed that the 18-2-9b2 LNP-treated group exhibited an increase in proteins responsible for various cellular processes, such as hematopoietic progenitor cell differentiation, trans- porter activity, and transmembrane transporter activity (FIG.20). These processes influence the efficacy, targeting, and intracellular processing for LNP-mediated delivery to the spleen. Representative proteins in Cluster 2, which are highly abundant in the corona, were selected and analyzed. It was shown that the amount of Abcb6, Itga2, Itga2b, and Itgb2 in the corona of the 18-2-9b2 LNP treatment group increased 2-5 times compared to these in MC3 and MC3 / 18PA LNP-treated groups (FIG.14E). Additionally, well-identified proteins like Apoe (liver-directed nanoparticle delivery) and Vtn (lung-directed nanoparticle delivery) were significantly decreased in the corona of the 18-2-9b2 LNP treated group, consistent with previous studies (FIGs.21A-21B). Specifically, Itga2b is predominantly expressed on the surface of platelets and megakaryocytes; consequently, nanoparticles that interact with Itga2b may be preferentially directed to the spleen due to its rich platelet content. Moreover, the 18-2-9b2 LNP was preincubated in Itga2b to evaluate the effect of protein adsorption on luciferase expression and intracellular uptake in RAW264.7 cells, which share many characteristics with splenic macrophages and serve as a useful model for studying their response to nanoparticles (FIG.14F). A 1.5-4.6 fold improvement in luciferase expression was observed across all amounts of protein tested (FIG.14G) and significantly higher intracellular uptake of Cy5-tagged mRNA compared to uncoated LNPs (FIG.14H). Collectively, these results demonstrated that endogenous targeting may behave as a potential mechanism for dendron-like design in facilitating mRNA delivery to the spleen. Example 5: 18-2-9b2 LNP-mediated targeted mRNA delivery to the splenic red pulp macrophage Having demonstrated that 18-2-9b2 LNP facilitates targeted mRNA delivery to the spleen, its ability to transfect specific cell types within the spleen was next evaluated. An Ai14 (constitutive loxP-STOP-loxP-tdTomato) reporter mouse model was used, which has been widely used for organ-specific gene editing studies. LNP-mediated intracellular delivery of Cre recombinase mRNA in this model deletes a flanking stop cassette, leading to tdTomato fluorescence expression within the transfected cell (FIG.4A). To benchmark the performance of lead LNP 18-2-9b2, a gold standard spleentropic MC3 / 18PA LNP was formulated as a positive control, in which 18PA shifts mRNA delivery of the FDA- approved MC3 LNP to the spleen (FIG.4B). Following a single systemic administration of - 84 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) 0.3 mg / kg Cre mRNA using 18-2-9b2 LNP, effective spleen gene modulation was observed 3 days post-administration (FIG.4C and FIGs.11A-11D). In particular, a transfection of 17.7% of red pulp macrophages (RPM) was observed, representing a 2.6-fold higher editing efficacy compared to MC3 / 18PA LNP. Immunostaining of tdTomato+ cells further validated the enhanced performance of 18-2-9b2 LNPs relative to MC3 / 18PA LNPs (FIG. 4D). Importantly, 18-2-9b2 LNP exhibited minimal in vivo toxicity, as evidenced by tissue section histology (FIG.12), showcasing its translation potential Red pulp macrophages (RPM), localizing in the splenic red pulp, play a crucial role in maintenance of blood homeostasis by actively phagocytosing injured and senescent erythrocytes and blood-borne particles. RPM accumulate large amounts of heme within their cytoplasm, and the transcription factor Spic has been reported to specifically regulate RPM development. Heme promotes degradation of the transcriptional repressor BTB and CNC homology 1 (BACH1), thereby reversing the repression of Spic by BACH1 (FIG.5A). Utilizing a Spic-GFP transgene mouse model, where the expression of Spic is inversely correlated with BACH1, and is easily quantifiable by GFP intensity in the RPM, it was demonstrated the translation potential of 18-2-9b2 LNP for therapeutic mRNA delivery to the spleen.18-2-9b2 LNP encapsulating BACH1 mRNA was systemic injected to Spic-GFP mice, and after 48 h, mice were sacrificed and the spleen was isolated for further evaluation. Quantitative PCR (qPCR) was employed to assess the delivery of BACH1 within the RPM (FIG.5B). In comparison to PBS-treated mice, the expression of BACH1 in 18-2-9b2 LNP-treated mice repressed the expression of Spic, leading to the downregulation of GFP signal observed through flow cytometry and immunostaining (FIGs.5C-5D and FIG.13). Collectively, these data demonstrated the development of 18-2-9b2 LNP with comb-like structure can facilitate targeted mRNA delivery to splenic RPM, and the delivery of therapeutic BACH1 mRNA using this platform holds potential to regulate transcriptional factors within splenic RPM for immunomodulatory applications. 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: - 85 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) (I), wherein: A is selected from the group consisting of and ; each occurrence of L1, if present, is consisting of -(optionally substituted C1-C alkylenyl)-, - 12-, - (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- C8 heteroarylenyl)-, -(optionally substituted C1-C12 alkylenyl)-X-, -(optionally substituted C2- C12 alkenylenyl)-X-, -(optionally substituted C1-C12 alkynylenyl)-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-; R1a, R1b, R1c, R1d, and R1e, if present, is independently selected from the group consisting of and optionally substituted C1-C6 alkyl, wherein: more than two of R1a, R1b, R1c, R1d, and R1eare C1-C6alkyl, 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-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 R3aand R3bis independently selected from the group consisting of optionally substituted C1-C24alkyl and optionally substituted C1-C24heteroalkyl; - 86 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) each occurrence of R4is independently selected from the group consisting of H and optionally substituted C1-C6alkyl; 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 and optionally substituted C1-C6alkyl. Embodiment 2 provides the compound of Embodiment 1, wherein each occurrence of L1is independently selected from the group consisting of -(CH2)1-5-, -(CH2)1-5CH(ORA)-, - (CH2)1-5O-, -(CH2)1-5N(RA)-, -(CH2)1-5N(R1e)-, and . 1 or 2, wherein -(L1)m- is -2-, -3-, -2N(CH3)(CH2)2-, - . of Embodiment 1, wherein R2is piperazinyl or C1-C6 alkyl substituted with at least one substituent selected from the group consisting of ORA, N(RA)(RB), pyrrolidinyl, piperidinyl, and piperazinyl. Embodiment 5 provides the compound of Embodiment 1 or 4, wherein R2is selected from the group consisting of -(CH2)2N(CH3)2, -(CH2)2N(CH2CH3)2, -(CH2)3N(CH2CH3)2, - the compound of formula (I) is selected from the group consisting of: , - 87 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) , , , each occurrence of RAand RBis independently selected from the group consisting of methyl and ethyl. Embodiment 8 provides the compound of any one of Embodiments 1-7, wherein each occurrence of L2is independently selected from the group consisting of -(CH2)1-2-, -O-, and - C(=O)-. Embodiment 9 provides the compound of any one of Embodiments 1-8, wherein - (L2)n- is -(CH2)2C(=O)O(CH2)-. Embodiment 10 provides the compound of any one of Embodiments 1-9, wherein R3aand R3bare each independently -(CH2)OC(=O)(C5-C20 alkyl), wherein the C5-C20 alkyl is optionally substituted with at least one C1-C3alkyl. Embodiment 11 provides the compound of any one of Embodiments 1-10, wherein - 88 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) R1a, R1b, R1c, R1d, and R1e, if present, are each , wherein: R5aand R5bare each independently -C(=O) ; and each occurrence of o is independently 1, 2, 3, 4, or 5; and p, q, and r are each independently 1, 2, or 3. Embodiment 12 provides the compound of Embodiment 11, wherein each occurrence of o is 2. Embodiment 13 provides the compound of Embodiment 11 or 12, wherein each occurrence of p, q, and r are is independently 1. Embodiment 14 provides the compound of any one of Embodiments 1-13, wherein each occurrence of R5aand R5bis independently selected from the group consisting of , , - 89 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) O O , 4 R is ethyl. Embodiment 16 provides the compound of any one of Embodiments 1-15, wherein R1a, R1b, R1c, R1d, and R1e, if present, are each independently selected from the group consisting of , - 90 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) , 5 , , - 91 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) . any one of Embodiments 1-16, which is 3-2-5, 4-2-5, 5-2-5, 6-2-5, 7-2-5, 8-2-5, 9- 2-5, 10-2-5, 11-2-5, 12-2-5, 13-2-5, 14-2-5, 15-2-5, 16-2-5, 17-2-5, 18-2-5, 1-2-6, 2-2-6, 3-2- 6, 4-2-6, 5-2-6, 6-2-6, 7-2-6, 8-2-6, 9-2-6, 10-2-6, 11-2-6, 12-2-6, 13-2-6, 14-2-6, 15-2-6, 16- 2-6, 17-2-6, 18-2-6, 1-2-6b, 2-2-6b, 3-2-6b, 4-2-6b, 5-2-6b, 6-2-6b, 7-2-6b, 8-2-6b, 9-2-6b, 10-2-6b, 11-2-6b, 12-2-6b, 13-2-6b, 14-2-6b, 15-2-6b, 16-2-6b, 17-2-6b, 18-2-6b, 1-2-7, 2-2- 7, 3-2-7, 4-2-7, 5-2-7, 6-2-7, 7-2-7, 8-2-7, 9-2-7, 10-2-7, 11-2-7, 12-2-7, 13-2-7, 14-2-7, 15- 2-7, 16-2-7, 17-2-7, 18-2-7, 1-2-7b, 2-2-7b, 3-2-7b, 4-2-7b, 5-2-7b, 6-2-7b, 7-2-7b, 8-2-7b, 9- 2-7b, 10-2-7b, 11-2-7b, 12-2-7b, 13-2-7b, 14-2-7b, 15-2-7b, 16-2-7b, 17-2-7b, 18-2-7b, 1-2- 7b2, 2-2-7b2, 3-2-7b2, 4-2-7b2, 5-2-7b2, 6-2-7b2, 7-2-7b2, 8-2-7b2, 9-2-7b2, 10-2-7b2, 11- 2-7b2, 12-2-7b2, 13-2-7b2, 14-2-7b2, 15-2-7b2, 16-2-7b2, 17-2-7b2, 18-2-7b2, 1-2-8, 2-2-8, 3-2-8, 4-2-8, 5-2-8, 6-2-8, 7-2-8, 8-2-8, 9-2-8, 10-2-8, 11-2-8, 12-2-8, 13-2-8, 14-2-8, 15-2-8, 16-2-8, 17-2-8, 18-2-8, 1-2-8b, 2-2-8b, 3-2-8b, 4-2-8b, 5-2-8b, 6-2-8b, 7-2-8b, 8-2-8b, 9-2- 8b, 10-2-8b, 11-2-8b, 12-2-8b, 13-2-8b, 14-2-8b, 15-2-8b, 16-2-8b, 17-2-8b, 18-2-8b, 1-2-9, 2-2-9, 3-2-9, 4-2-9, 5-2-9, 6-2-9, 7-2-9, 8-2-9, 9-2-9, 10-2-9, 11-2-9, 12-2-9, 13-2-9, 14-2-9, 15-2-9, 16-2-9, 17-2-9, 18-2-9, 1-2-9b, 2-2-9b, 3-2-9b, 4-2-9b, 5-2-9b, 6-2-9b, 7-2-9b, 8-2- 9b, 9-2-9b, 10-2-9b, 11-2-9b, 12-2-9b, 13-2-9b, 14-2-9b, 15-2-9b, 16-2-9b, 17-2-9b, 18-2-9b, 1-2-9b2, 2-2-9b2, 3-2-9b2, 4-2-9b2, 5-2-9b2, 6-2-9b2, 7-2-9b2, 8-2-9b2, 9-2-9b2, 10-2-9b2, 11-2-9b2, 12-2-9b2, 13-2-9b2, 14-2-9b2, 15-2-9b2, 16-2-9b2, 17-2-9b2, 18-2-9b2, 1-2-9b3, 2-2-9b3, 3-2-9b3, 4-2-9b3, 5-2-9b3, 6-2-9b3, 7-2-9b3, 8-2-9b3, 9-2-9b3, 10-2-9b3, 11-2-9b3, 12-2-9b3, 13-2-9b3, 14-2-9b3, 15-2-9b3, 16-2-9b3, 17-2-9b3, 18-2-9b3, 1-2-9v, 2-2-9v, 3-2- 9v, 4-2-9v, 5-2-9v, 6-2-9v, 7-2-9v, 8-2-9v, 9-2-9v, 10-2-9v, 11-2-9v, 12-2-9v, 13-2-9v, 14-2- 9v, 15-2-9v, 16-2-9v, 17-2-9v, 18-2-9v, 1-2-10, 2-2-10, 3-2-10, 4-2-10, 5-2-10, 6-2-10, 7-2- 10, 8-2-10, 9-2-10, 10-2-10, 11-2-10, 12-2-10, 13-2-10, 14-2-10, 15-2-10, 16-2-10, 17-2-10, 18-2-10, 1-2-11, 2-2-11, 3-2-11, 4-2-11, 5-2-11, 6-2-11, 7-2-11, 8-2-11, 9-2-11, 10-2-11, 11- 2-11, 12-2-11, 13-2-11, 14-2-11, 15-2-11, 16-2-11, 17-2-11, and 18-2-11. Embodiment 18 provides the compound of any one of Embodiments 1-17, wherein the compound of formula (I) is: - 92 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) , a (a) at least one ionizable lipid comprising the compound of any one of Embodiments 1-18; (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 20 provides the LNP of Embodiment 19, 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 21 provides the LNP of Embodiment 19 or 20, wherein the at least ionizable lipid compound comprises or consists essentially of: , 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 23 provides the LNP of any one of Embodiments 19-22, wherein the neutral lipid comprises or consists essentially of at least one neutral lipid selected from the - 93 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) group consisting of dioleoylphosphatidylethanolamine (DOPE), distearoylphosphatidylcholine (DSPC), and dioleoylphosphatidylcholine (DOPC), optionally wherein the neutral lipid comprises or consists essentially of dioleoylphosphatidylethanolamine (DOPE). Embodiment 24 provides the LNP of any one of Embodiments 19-23, 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 25 provides the LNP of any one of Embodiments 19-24, wherein the at least one cholesterol lipid and / or modified derivative thereof comprises or consists essentially of cholesterol. Embodiment 26 provides the LNP of any one of Embodiments 19-25, 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 27 provides the LNP of any one of Embodiments 19-26, 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 28 provides the LNP of any one of Embodiments 19-27, wherein the LNP has a molar ratio of (a) : (b) : (c) : (d) of about 35:16:46.5:2.5. Embodiment 29 provides the LNP of any one of Embodiments 19-28, 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 30 provides the LNP of Embodiment 29, 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 31 provides the LNP of Embodiment 29 or 30, wherein the cargo molecule comprises a nucleic acid. Embodiment 32 provides the LNP of Embodiment 31, wherein the nucleic acid is DNA or RNA. Embodiment 33 provides the LNP of Embodiment 31 or 32, 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 34 provides the LNP of Embodiment 33, wherein the mRNA comprises BACH1 mRNA. - 94 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) Embodiment 35 provides the LNP of Embodiment 33, wherein the mRNA encodes an enzyme. Embodiment 36 provides the LNP of Embodiment 33, wherein the mRNA encodes a clustered regularly interspaced short palindrome repeats (CRISPR) associated protein, optionally wherein the CRISPR associated protein is Cas9. Embodiment 37 provides the LNP of Embodiment 36, wherein the nucleic acid cargo further comprises sgRNA. Embodiment 38 provides a pharmaceutical composition comprising the lipid nanoparticle (LNP) of any one of Embodiments 19-37 and at least one pharmaceutically acceptable carrier. Embodiment 39 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 29-37. Embodiment 40 provides the method of Embodiment 39, wherein the disease is at least one selected from the group consisting of cancer, cardiovascular disease, and a metabolic disease, optionally wherein the disease relates to aberrant red blood cell differentiation. Embodiment 41 provides the method of Embodiment 39 or 40, wherein the disease is associated with BACH1 overexpression. Embodiment 42 provides a method of delivering at least one therapeutic cargo molecule to a subject’s spleen, the method comprising administering to the subject at least one lipid nanoparticle (LNP) of any one of Embodiments 29-37. Embodiment 43 provides the method of Embodiment 42, wherein delivery of the at least one therapeutic cargo molecule is selective for the spleen, optionally wherein the at least one therapeutic cargo is not substantially delivered to the liver, heart, lungs, and / or kidney. Embodiment 44 provides the method of Embodiment 42 or 43, wherein the at least one therapeutic cargo molecule is delivered to a splenic red pulp macrophage. Embodiment 45 provides the method of any one of Embodiments 42-44, wherein 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. Embodiment 46 provides the method of any one of Embodiments 42-45, wherein the therapeutic cargo molecule comprises a nucleic acid. Embodiment 47 provides the method of Embodiment 46, wherein the nucleic acid is DNA or RNA. - 95 - 56556621.1 Attorney Docket No.046483-7480WO1(04054) Embodiment 48 provides the method of Embodiment 46 or 47, 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 49 provides the method of Embodiment 48, wherein the mRNA comprises BACH1 mRNA. Embodiment 50 provides the method of Embodiment 48, wherein the mRNA encodes an enzyme. Embodiment 51 provides the method of Embodiment 48, wherein the mRNA encodes a clustered regularly interspaced short palindrome repeats (CRISPR) associated protein, optionally wherein the CRISPR associated protein is Cas9. Embodiment 52 provides the method of Embodiment 51, wherein the nucleic acid cargo further comprises sgRNA. 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. - 96 - 56556621.1

Claims

Attorney Docket No.046483-7480WO1(04054) 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 ; each occurrence of L1, if present, is consistingof -(optionally substituted C1-C12 alkylenyl)-, - -, - (optionally substituted C1-C12 alkynylenyl)-, -(optionally substituted C1-C12 heteroalkylenyl)- , -(optionally substituted C3-C8 cycloalkylenyl)-, -(optionally substituted C2-C8 heterocyloalkylenyl)-, -(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-; R1a, R1b, R1c, R1d, and R1e, if present, is independently selected from the group consisting of and optionally substituted C1-C6alkyl, wherein: no more than two of R1a, R1b, R1c, R1d, and R1eare C1-C6 alkyl, 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 - - 97 - 56556621.1Attorney Docket No.046483-7480WO1(04054) (optionally substituted C1-C6 alkylenyl)-, -C(=O)-, -O-, and -N(RA)-; each occurrence of R3aand R3bis independently selected from the group consisting of optionally substituted C1-C24 alkyl and optionally substituted C1-C24 heteroalkyl; each occurrence of R4is independently selected from the group consisting of H and optionally substituted C1-C6 alkyl; 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 and optionally substituted C1-C6 alkyl.

2. The compound of claim 1, 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)-, - (CH2)1-5N(R1e)-, and .

3. The compound of claim 1 or 2, wherein -(L1)m- is selected from the group consisting of -(CH2)2-, -(CH2)3-, -(CH2)2N(CH3)(CH2)2-, -(CH2)3N(CH3)(CH2)3-,4. The compound of claim 1, wherein R2is piperazinyl or C1-C6alkyl substituted with at least one substituent selected from the group consisting of ORA, N(RA)(RB), pyrrolidinyl, piperidinyl, and piperazinyl.

5. The compound of claim 1 or 4, wherein R2is selected from the group consisting of - ,- 98 - 56556621.1Attorney Docket No.046483-7480WO1(04054) .of formula (I) is selected from the group consisting of: RAN R1a, ,7. The compound of any one of claims 1-6, wherein each occurrence of RAand RBis independently selected from the group consisting of methyl and ethyl.

8. The compound of any one of claims 1-7, wherein each occurrence of L2is - 99 - 56556621.1Attorney Docket No.046483-7480WO1(04054) independently selected from the group consisting of -(CH2)1-2-, -O-, and -C(=O)-.

9. The compound of any one of claims 1-8, wherein -(L2)n- is -(CH2)2C(=O)O(CH2)-.

10. The compound of any one of claims 1-9, wherein R3aand R3bare each independently - (CH2)OC(=O)(C5-C20alkyl), wherein the C5-C20alkyl is optionally substituted with at least one C1-C3 alkyl.

11. The compound of any one of claims 1-10, wherein R1a, R1b, R1c, R1d, and R1e, if present, are each , wherein: R5aand R5bare each substituted C5-C20alkyl); andeach occurrence of o is independently 1, 2, 3, 4, or 5; and p, q, and r are each independently 1, 2, or 3.

12. The compound of claim 11, wherein each occurrence of o is 2.

13. The compound of claim 11 or 12, wherein each occurrence of p, q, and r are is independently 1.

14. The compound of any one of claims 1-13, wherein each occurrence of R5aand R5bis independently selected from the group consisting of , ,- 100 - 56556621.1Attorney Docket No.046483-7480WO1(04054) , , ,- 101 - 56556621.1Attorney Docket No.046483-7480WO1(04054) .

16. The compound of any one of claims 1-15, wherein R1a, R1b, R1c, R1d, and R1e, if present, are each independently selected from the group consisting of ,- 102 - 56556621.1Attorney Docket No.046483-7480WO1(04054) , ,17. The compound of any one of claims 1-16, which is selected from the group consisting of 1-2-5, 2-2-5, 3-2-5, 4-2-5, 5-2-5, 6-2-5, 7-2-5, 8-2-5, 9-2-5, 10-2-5, 11-2-5, 12-2-5, 13-2-5, 14-2-5, 15-2-5, 16-2-5, 17-2-5, 18-2-5, 1-2-6, 2-2-6, 3-2-6, 4-2-6, 5-2-6, 6-2-6, 7-2-6, 8-2-6, 9-2-6, 10-2-6, 11-2-6, 12-2-6, 13-2-6, 14-2-6, 15-2-6, 16-2-6, 17-2-6, 18-2-6, 1-2-6b, 2-2-6b, 3-2-6b, 4-2-6b, 5-2-6b, 6-2-6b, 7-2-6b, 8-2-6b, 9-2-6b, 10-2-6b, 11-2-6b, 12-2-6b, 13-2-6b, 14-2-6b, 15-2-6b, 16-2-6b, 17-2-6b, 18-2-6b, 1-2-7, 2-2-7, 3-2-7, 4-2-7, 5-2-7, 6-2-7, 7-2-7, 8-2-7, 9-2-7, 10-2-7, 11-2-7, 12-2-7, 13-2-7, 14-2-7, 15-2-7, 16-2-7, 17-2-7, 18-2-7, 1-2-7b, 2-2-7b, 3-2-7b, 4-2-7b, 5-2-7b, 6-2-7b, 7-2-7b, 8-2-7b, 9-2-7b, 10-2-7b, 11-2-7b, 12-2-7b, 13-2-7b, 14-2-7b, 15-2-7b, 16-2-7b, 17-2-7b, 18-2-7b, 1-2-7b2, 2-2-7b2, 3-2-7b2, 4-2-7b2, 5- 2-7b2, 6-2-7b2, 7-2-7b2, 8-2-7b2, 9-2-7b2, 10-2-7b2, 11-2-7b2, 12-2-7b2, 13-2-7b2, 14-2- 7b2, 15-2-7b2, 16-2-7b2, 17-2-7b2, 18-2-7b2, 1-2-8, 2-2-8, 3-2-8, 4-2-8, 5-2-8, 6-2-8, 7-2-8, 8-2-8, 9-2-8, 10-2-8, 11-2-8, 12-2-8, 13-2-8, 14-2-8, 15-2-8, 16-2-8, 17-2-8, 18-2-8, 1-2-8b, 2-2-8b, 3-2-8b, 4-2-8b, 5-2-8b, 6-2-8b, 7-2-8b, 8-2-8b, 9-2-8b, 10-2-8b, 11-2-8b, 12-2-8b, 13-2-8b, 14-2-8b, 15-2-8b, 16-2-8b, 17-2-8b, 18-2-8b, 1-2-9, 2-2-9, 3-2-9, 4-2-9, 5-2-9, 6-2- - 103 - 56556621.1Attorney Docket No.046483-7480WO1(04054) 9, 7-2-9, 8-2-9, 9-2-9, 10-2-9, 11-2-9, 12-2-9, 13-2-9, 14-2-9, 15-2-9, 16-2-9, 17-2-9, 18-2-9, 1-2-9b, 2-2-9b, 3-2-9b, 4-2-9b, 5-2-9b, 6-2-9b, 7-2-9b, 8-2-9b, 9-2-9b, 10-2-9b, 11-2-9b, 12- 2-9b, 13-2-9b, 14-2-9b, 15-2-9b, 16-2-9b, 17-2-9b, 18-2-9b, 1-2-9b2, 2-2-9b2, 3-2-9b2, 4-2- 9b2, 5-2-9b2, 6-2-9b2, 7-2-9b2, 8-2-9b2, 9-2-9b2, 10-2-9b2, 11-2-9b2, 12-2-9b2, 13-2-9b2, 14-2-9b2, 15-2-9b2, 16-2-9b2, 17-2-9b2, 18-2-9b2, 1-2-9b3, 2-2-9b3, 3-2-9b3, 4-2-9b3, 5-2- 9b3, 6-2-9b3, 7-2-9b3, 8-2-9b3, 9-2-9b3, 10-2-9b3, 11-2-9b3, 12-2-9b3, 13-2-9b3, 14-2-9b3, 15-2-9b3, 16-2-9b3, 17-2-9b3, 18-2-9b3, 1-2-9v, 2-2-9v, 3-2-9v, 4-2-9v, 5-2-9v, 6-2-9v, 7-2- 9v, 8-2-9v, 9-2-9v, 10-2-9v, 11-2-9v, 12-2-9v, 13-2-9v, 14-2-9v, 15-2-9v, 16-2-9v, 17-2-9v, 18-2-9v, 1-2-10, 2-2-10, 3-2-10, 4-2-10, 5-2-10, 6-2-10, 7-2-10, 8-2-10, 9-2-10, 10-2-10, 11- 2-10, 12-2-10, 13-2-10, 14-2-10, 15-2-10, 16-2-10, 17-2-10, 18-2-10, 1-2-11, 2-2-11, 3-2-11, 4-2-11, 5-2-11, 6-2-11, 7-2-11, 8-2-11, 9-2-11, 10-2-11, 11-2-11, 12-2-11, 13-2-11, 14-2-11, 15-2-11, 16-2-11, 17-2-11, and 18-2-11.

18. The compound of any one of claims 1-17, wherein the compound of formula (I) is: ,19. A lipid nanoparticle (LNP) comprising: (a) at least one ionizable lipid comprising the compound of any one of claims 1- 18; (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.

20. The LNP of claim 19, 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. - 104 - 56556621.1Attorney Docket No.046483-7480WO1(04054) 21. The LNP of claim 19 or 20, wherein the at least ionizable lipid compound comprises or consists essentially of: ,22. The LNP of any one of claims 19-21, 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.

23. The LNP of any one of claims 19-22, 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).

24. The LNP of any one of claims 19-23, 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.

25. The LNP of any one of claims 19-24, wherein the at least one cholesterol lipid and / or modified derivative thereof comprises or consists essentially of cholesterol.

26. The LNP of any one of claims 19-25, 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%. - 105 - 56556621.1Attorney Docket No.046483-7480WO1(04054) 27. The LNP of any one of claims 19-26, 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).

28. The LNP of any one of claims 19-27, wherein the LNP has a molar ratio of (a) : (b) : (c) : (d) of about 35:16:46.5:2.

5.

29. The LNP of any one of claims 19-28, 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.

30. The LNP of claim 29, 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.

31. The LNP of claim 29 or 30, wherein the cargo molecule comprises a nucleic acid.

32. The LNP of claim 31, wherein the nucleic acid is DNA or RNA.

33. The LNP of claim 31 or 32, 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.

34. The LNP of claim 33, wherein the mRNA comprises BACH1 mRNA.

35. The LNP of claim 33, wherein the mRNA encodes an enzyme.

36. The LNP of claim 33, wherein the mRNA encodes a clustered regularly interspaced short palindrome repeats (CRISPR) associated protein, optionally wherein the CRISPR associated protein is Cas9.

37. The LNP of claim 36, wherein the nucleic acid cargo further comprises sgRNA. - 106 - 56556621.1Attorney Docket No.046483-7480WO1(04054) 38. A pharmaceutical composition comprising the lipid nanoparticle (LNP) of any one of claims 19-37 and at least one pharmaceutically acceptable carrier.

39. 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 claims 29-37.

40. The method of claim 39, wherein the disease is at least one selected from the group consisting of cancer, cardiovascular disease, and a metabolic disease, optionally wherein the disease relates to aberrant red blood cell differentiation.

41. The method of claim 39 or 40, wherein the disease is associated with BACH1 overexpression.

42. A method of delivering at least one therapeutic cargo molecule to a subject’s spleen, the method comprising administering to the subject at least one lipid nanoparticle (LNP) of any one of claims 29-37.

43. The method of claim 42, wherein delivery of the at least one therapeutic cargo molecule is selective for the spleen, optionally wherein the at least one therapeutic cargo is not substantially delivered to the liver, heart, lungs, and / or kidney.

44. The method of claim 42 or 43, wherein the at least one therapeutic cargo molecule is delivered to a splenic red pulp macrophage.

45. The method of any one of claims 42-44, wherein 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.

46. The method of any one of claims 42-45, wherein the therapeutic cargo molecule comprises a nucleic acid.

47. The method of claim 46, wherein the nucleic acid is DNA or RNA. - 107 - 56556621.1Attorney Docket No.046483-7480WO1(04054) 48. The method of claim 46 or 47, 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.

49. The method of claim 48, wherein the mRNA comprises BACH1 mRNA.

50. The method of claim 48, wherein the mRNA encodes an enzyme.

51. The method of claim 48, wherein the mRNA encodes a clustered regularly interspaced short palindrome repeats (CRISPR) associated protein, optionally wherein the CRISPR associated protein is Cas9.

52. The method of claim 51, wherein the nucleic acid cargo further comprises sgRNA. - 108 - 56556621.1

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