Degradable, branched lipid compounds, methods of preparation thereof, lipid nanoparticles (LNPS) comprising the same, and methods of use thereof

Abstract

The present disclosure relates, in one aspect, to ionizable lipid compounds of formula (I). In another aspect, the present disclosure relates to the preparation of ionizable lipids of formula (I). In another aspect, the present disclosure relates to lipid nanoparticles (LNPs) comprising at least one ionizable lipid of formula (I), and methods of use thereof.

Description

[0001]Attorney Docket No.046483-7436WO1(03708) TITLE OF THE INVENTION Degradable, Branched Lipid Compounds, Methods of Preparation Thereof, Lipid Nanoparticles (LNPs) Comprising the Same, and Methods of Use Thereof 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. CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority under 35 U.S.C. § 119(e) to U.S. Provisional Patent Application No.63 / 581,832, filed September 11, 2023, which is incorporated herein by reference in its entirety. BACKGROUND Messenger RNA (mRNA) technology has enabled breakthroughs in the treatment and prevention of a variety of pathological conditions, including cancers, infectious diseases, metabolic disorders, and congenital diseases. Indeed, mRNA-based therapeutics have achieved tremendous clinical success in vaccines, protein replacement therapies, and gene editing therapies. However, the unfavorable physicochemical properties of mRNA (e.g., large size, negative charge, and structural instability) make it ineffective in its native form, necessitating a carrier for efficient intracellular delivery. Lipid nanoparticles (LNPs) represent the most clinically advanced, non-viral platform for mRNA delivery. LNPs typically comprise one or more ionizable lipids (or lipidoids), one or more phospholipids, cholesterol, or derivatives thereof, and one or more polymer- conjugated lipids (e.g., polyethylene glycol (PEG)-conjugated lipids). Lipidoids play a key role in protecting and transporting mRNA cargo. At acidic pH, lipidoids are positively charged, allowing them to both condense mRNA during LNP formulation and disrupt the endosomal membrane during cellular internalization. At physiological pH, lipidoids maintain a neutral charge, limiting toxicity and improving LNP pharmacokinetic properties. Lipidoids typically comprise an ionizable headgroup and two or more aliphatic substituents (e.g., alkyl tails) that are connected to the ionizable headgroup by linkers. Notably, degradable linkers, which may comprise ester linkages, are preferentially used in - 1 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) lipidoids to allow for hydrolysis in vivo with improved biocompatibility. In addition, branched alkyl tails are favored, since these tails can promote the lipidoid to adopt a cone- shape structure that is beneficial for endosomal escape and mRNA delivery. However, the synthesis of degradable lipidoids comprising extended, branched alkyl groups is challenging due to the lack of commercially available branched building blocks, which hinders systematic optimization and investigation thereof. Historically, branched lipidoids were exclusively constructed using two main steps: first, the preparation of a branched tail intermediate containing a functional group (e.g., bromide, aldehyde, acrylate, hydroxyl, and / or carboxyl group, inter alia); second, the connection of branched tail(s) to the headgroup. This construction method typically involves tedious synthesis and purification, making it difficult to systemically optimize these chemical structures and investigate structure-activity relationships (SARs). There is thus a need in the art for degradable lipids that can be prepared in a high- throughput manner, lipid nanoparticles comprising the same, and methods of use thereof for the treatment and / or prevention of a variety of pathological conditions. The present disclosure addresses this need. BRIEF SUMMARY In one aspect, the disclosure provides a compound formula (I), or a salt or solvate thereof: (I), herein. comprising at least one ionizable lipid compound of formula (I), at least one helper lipid, at least one cholesterol lipid, and at least one polymer-conjugated lipid and / or a modified derivative thereof. In another aspect, the disclosure provides a method of preparing the ionizable lipid compound of formula (I): (I), - 2 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) the method comprising: (a) contacting an amine compound of formula (A1) or (A2): (A1) or (A2), and at least , to form an alkoxide (b) contacting the an activated acid compound of formula (C): , wherein X is a and wherein A, L, R3b, R4a, R4b, R5a, R5b, and R6are defined In another aspect, the disclosure provides a pharmaceutical composition comprising the lipid nanoparticle of the disclosure and at least one pharmaceutically acceptable carrier. In another aspect, the disclosure provides a method of treating, preventing, and / or ameliorating a disease or disorder in a subject, the method comprising administering to the subject at least one lipid nanoparticle (LNP). 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-1B: illustrative space shuttle-inspired design of SRB-lipidoids for mRNA delivery. In NASA’s space launch system, two detachable solid rocket boosters are appended to provide transient thrust to deliver the space shuttle into space. Inspired by this design, a structure- and function-mimetic strategy was adopted: two branch tails are appended to an inefficacious lipidoid in situ via degradable linkers, endowing transient potency of SRB- lipidoids to facilitate intracellular transport of mRNA. FIGs.2A-2I: construction of SRB-lipidoids and screening of Library 1. FIG.2A: synthetic scheme depicting the tandem, hierarchical, and in situ construction of certain exemplary SRB-lipidoids described in the present application, based on a one-pot, two-step, - 3 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) three-component reaction (3-CR). An amine reacts with excessive alkyl epoxide and the resulting aminoalcohol lipidoid further reacts with excessive acyl chloride to afford SRB- lipidoid. FIG.2B: depicts the workflow for the synthesis and evaluation of Library 1. FIG. 2C: in vitro mLuc expression shown in a heat map (n = 2). FIG.2D: in vivo mLuc expression shown in a heat map (n = 2). FIG.2E: correlation between in vitro and in vivo results. The black dashed line indicates 10,000 units in vitro. The blue dashed line indicates the performance of MC3 LNP in vivo. FIG.2F: body tail-activity relationship. FIG.2G: branch tail-activity relationship. FIG.2H: total carbon number-activity relationship. FIG.2I: total carbon number-symmetry-activity relationship. The grey shadows indicate background levels. FIG.3: cell viability of aminoalcohol lipidoid-formulated LNPs and SRB-LNPs in Library 1. HepG2 cells were treated with various LNPs at a dose of 15 ng mRNA / well for 24 h. No major cytotoxicity was induced by LNPs. The dashed line indicates 80% cell viability. Data are presented as mean ± SD (n = 2). FIG.4: Representative results of in vivo and ex vivo bioluminescence imaging (BLI) during LNP screening. Mice were i.v. injected with mLuc-loaded SRB-LNPs at an mRNA dose of 0.1 mg / kg. Images were taken at 4 h post-treatment. The signal mainly localized in the upper abdomen based on in vivo BLI, which was identified to come from the liver by ex vivo BLI. FIGs.5A-5D: Comparison of purified 1-10-8(1) and 1-10-8(2). FIG.5A: synthetic route for 1-10-8(1). FIG.5B: mRNA encapsulation efficiencies of 1-10-8(2) LNP and 1-10- 8(1) LNP. FIGs.5C-5D: in vitro mLuc expression (n = 2). The grey shadow indicates background level. Lipidoids were purified using a CombiFlash NextGen 300+ chromatography system, and the desired fractions were collected and confirmed by mass spectrum. LNPs were formulated by pipette mixing. FIGs.6A-6F: optimization of headgroup and screening of Library 2. FIG.6A: the workflow for the synthesis and evaluation of Library 2. FIG.6B: in vitro mLuc expression (n = 2). FIG.6C: in vivo mLuc expression (n = 2). Efficacious SRB-lipidoids and their amines are highlighted in red. The grey shadow indicates background level. FIG.6D: correlation between in vitro and in vivo results. The black dashed line indicates 10,000 units in vitro. The blue dashed line indicates the performance of MC3 LNP in vivo. FIG.6E: structures of efficacious amines and SRB-lipidoids. The structure of lead SRB-lipidoid 11-10-8 is shown. FIG.6F: a space-filling model of the 11-10-8 SRB-lipidoid. It possesses a space shuttle- resembling structure with a total carbon number of 18, a symmetry of 1, and a packing - 4 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) parameter (P) of 4.1 FIG.7: Cell viability of SRB-LNPs in Library 2. HepG2 cells were treated with various LNPs at a dose of 15 ng mRNA / well for 24 h. No major cytotoxicity was induced by SRB-LNPs. The blue dot indicates MC3 LNP. The dashed line indicates 80% cell viability. Data are presented as mean ± SD (n = 2). FIG.8: Structural illustration of 11-10-8 and MC3. Critical packing parameters were calculated based on molecular dynamics simulations. The dimensionless packing parameter P of a lipid molecule was calculated as P = V / (AL) based on its Van der Waals molecule volume (V), cross section area of polar head (A) and average tail length (L). FIGs.9A-9E: Prediction, optimization, and characterization of SRB-LNPs. FIG.9A: prediction and verification of in vivo performance for unidentified SRB-lipidoids based on structural criteria (n = 2). In vivo performance of unidentified SRB-lipidoids was verified at 4 h post-treatment of mLuc-loaded SRB-LNPs at an mRNA dose of 0.1 mg / kg. The red dashed line indicates the performance of 11-10-8 LNP, while the blue dashed line indicates the performance of MC3 LNP. FIG.9B: optimization of 11-10-8 LNP formulation (n = 2). Mice were injected i.v. with mLuc-loaded LNPs at an mRNA dose of 0.1 mg / kg. Images were taken at 4 h post-treatment. FIG.9C: cryo-EM image of 11-10-8 LNP. Scale bar = 50 nm. FIG.9D: physiochemical parameters of 11-10-8 LNP (n = 3). FIG.9E: inhibition of 11-10-8 LNP uptake by various endocytic inhibitors (n = 3). Amiloride is an inhibitor of macropinocytosis; Chlorpromazine is an inhibitor of clathrin-mediated endocytosis; Genistein is an inhibitor of caveolae-mediated endocytosis; Methyl-β-cyclodextrin (Mβ-CD) is an inhibitor of lipid rafter-mediated endocytosis. Data are presented as mean ± SD. FIG.10: TNS assay was used to determine the apparent pKaof 11-10-8 LNP. TNS fluorescence signal corresponds to ionization. pKa is calculated as the pH corresponding to half of the maximum TNS fluorescence value. FIG.11: Hemolysis of 11-10-8 LNP at pH 7.4 or 6.0. RBCs were incubated with 11- 10-8 LNP at an mRNA concentration of 3 μg / mL at 37 °C for 1 h. Positive and negative controls were carried out with 0.1% Triton-X and 1× PBS, respectively. Data are presented as mean ± SD (n = 3). FIGs.12A-12B: 11-10-8 LNP-mediated in vitro mLuc delivery and cytotoxicity. FIG. 12A: in vitro mLuc expression. FIG.12B: cell viability; HepG2 cells were treated with 11- 10-8 LNP at indicated doses for 24 h. Data are presented as mean ± SD (n = 3). FIGs.13A-13D: Flow cytometry analysis of GFP mRNA expression in HepG2 cells. FIG.13A: gating strategy. FIG.13B: representative results of GFP expression. FIG.13C: - 5 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) mean fluorescence intensity (MFI) of cells. FIG.13D: percentage of GFP-positive cells. HepG2 cells were treated with GFP mRNA-loaded 11-10-8 LNP at the indicated doses for 24 h. Data are presented as mean ± SD (n = 3). FIGs.14A-14E: SRB-LNP-mediated hepatic delivery of mRNA-based therapeutics and gene editors. FIG.14A: ex vivo BLI of major organs from treated mice and their quantification (n = 3). Mice were i.v. injected with mLuc-loaded LNPs at an mRNA dose of 0.1 mg / kg. Images were taken at 4 h post-treatment. FIG.14B: ex vivo fluorescence imaging of major organs from treated mice and their quantification (n = 3). Mice were i.v. injected with GFP mRNA-loaded LNPs at an mRNA dose of 0.25 mg / kg. Images were taken at 4 h post-treatment. FIG.14C: LNP-mediated FGF21 mRNA delivery (n = 4). Mice were i.v. injected with FGF21 mRNA-loaded LNPs at an mRNA dose of 0.25 mg / kg. Serum was collected at the indicated time points for ELISA analysis of serum FGF21. FIGs.14D-14E: LNP-mediated Cas9 mRNA / TTR sgRNA co-delivery and gene editing. Mice were i.v. injected with LNPs co-delivering Cas9 mRNA / TTR sgRNA (4:1, wt:wt) at a total RNA dose of 1 mg / kg. Mice were euthanized on day 7, and DNA was extracted from the liver to determine on-target indel frequency by next-generation sequencing (FIG.14D, n = 3). Serum was collected at the indicated time points for ELISA analysis of serum TTR (FIG.14E, n = 3). Data are presented as mean ± SD. FIGs.15A-15B: ALT and AST analysis. FIG.15A: serum ALT (n = 3). FIG.15B: serum AST (n = 3). Mice were i.v. injected with LNPs co-delivering Cas9 mRNA / TTR sgRNA (4:1, wt:wt) at a total RNA dose of 1 mg / kg. Serum was collected for ALT and AST analysis at 24 h post-treatment. FIGs.16A-16B: in vivo LNP-mediated TTR siRNA delivery. FIG.16A: dose- dependent TTR silencing (n = 3). Mice were i.v. injected with TTR siRNA-loaded LNPs at different doses. Serum was collected on day 3 for ELISA analysis of serum TTR. FIG.16B: duration of TTR silencing (n = 3). Mice were i.v. injected with TTR siRNA-loaded LNPs at a dose of 1 mg / kg. Serum was collected at indicated time points for ELISA analysis of serum TTR. Data are presented as mean ± SD. FIGs.17A-17F: DB-LNP-mediated hepatic delivery of FGF21 mRNA; LNP- mediated FGF21 mRNA delivery (n = 4 biologically independent samples). Male mice were fed a HFD for three weeks to induce obesity (body weight ~30g) and fatty liver. These obese mice were i.v. injected with FGF21 mRNA-loaded LNPs at an mRNA dose of 0.25 mg / kg. Serum was collected at the indicated time points for ELISA analysis of FGF21. AUC of FGF21 exposure during the time interval 0–48 h was determined. Statistical significance was - 6 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) evaluated by an unpaired two-tailed Student’s t-test. FIG.17A: A scheme of FGF21 mRNA therapy in HFD-induced obese mice. Obese mice were i.v. injected with various LNP formulations at an mRNA dose of 0.25 mg / kg every other day for three doses. Male mice fed a normal chow diet (NCD) was used a control group. FIG.17B: Body weight growth curve (n = 4 biologically independent samples). FIG.17C: Body weight on Day 6 (n = 4 biologically independent samples). Statistical significance was evaluated by a one-way ANOVA with Tukey’s correction. FIG.17D: Liver weight (n = 4 biologically independent samples). Statistical significance was evaluated by a one-way ANOVA with Tukey’s correction. FIG. 17E: Representative images of liver histological examinations (H&E staining). Scale bars: 100 μm. FIG.17F: Liver steatosis score (n = 12). A total of 12 images for each group (three random fields for each liver section, four mice per group) were analyzed semi-quantitatively for liver steatosis. Statistical significance was evaluated by a one-way ANOVA with Tukey’s correction. Data are presented as mean ± SD. FIGs.18A-18B: Cell viability and IC50of DB-LNPs in Library 1 and other LNPs. FIG.18A: Cell viability. HepG2 cells were treated with LNPs at 15 ng mRNA / well (0.24 nM), 50 ng mRNA / well (0.8 nM) or 200 ng mRNA / well (3.2 nM) for 24 h. No obvious cytotoxicity was observed for all LNPs at a low dose (i.e., 15 ng mRNA / well). The dashed line indicates 80% cell viability. Data are presented as mean ± SD (n = 3 biologically independent samples). FIG.18B: Half-maximal inhibitory concentration (IC50). IC50was determined by nonlinear regression of dose and cell viability using GraphPad Prism 8.0. FIG.19: Cell viability of DB-LNPs in Library 2. HepG2 cells were treated with various LNPs at a dose of 15 ng mRNA / well (0.24 nM) for 24 h. No obvious cytotoxicity was induced by DB-LNPs. The dashed line indicates 80% cell viability. Data are presented as mean ± SD (n = 3 biologically independent samples). FIG.20:1H-NMR spectrum of purified 11-10-8 SRB-lipidoid in MeOD. FIG.21: TNS assay was used to determine the apparent pKa of 11-10-8 LNP. TNS fluorescence signal corresponds to ionization. pKais calculated as the pH corresponding to half of the maximum TNS fluorescence value. FIGs.22A-22C: GFP expression in the liver at 4 h post-treatment of GFP mRNA- loaded LNPs. Mice were i.v. injected with GFP mRNA-loaded LNPs, 11-10-8 LNP (FIG. 22B), MC3 LNP (FIG.22C) or PBS only (FIG.22A); at an mRNA dose of 0.25mg / kg. Livers were collected and cryo-sectioned for immunofluorescence staining at 4 h post- treatment. Kupffer cells (F4 / 80+) and liver sinusoidal endothelial cells (VE-Cad+) were stained, respectively. Representative images from three independent experiments were - 7 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) shown. Scale bars: 100 μm. FIG.23: In vivo mLuc expression after i.m. injection of LNPs. Mice were i.m. injected with mLuc-loaded LNPs at an mRNA dose of 0.1 mg / kg. Images were taken at 4 h post-treatment. Data are presented as mean ± SD (n = 3 biologically independent samples). Statistical significance was evaluated by a one-way ANOVA with Tukey’s correction. 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, 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 - 8 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 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 In one aspect, the present disclosure describes the development of novel synthetic approaches that enable rapid, cost-efficient, and high-throughput synthesis of degradable branched lipidoids using readily available building blocks. In another aspect, the present disclosure describes the design of degradable, branched lipidoids. In certain embodiments, the present disclosure finds inspiration in a non-limiting manner in the structure and function of the National Aeronautics and Space Administration (NASA)’s space launch system (FIGs.1A-1B), in which two detachable solid rocket boosters are appended to provide transient thrust and transport the space shuttle into space. In certain embodiments, a novel construction strategy was devised for tandem, hierarchical, and in situ assembly of space shuttle-resembling branched (SRB) lipidoids based on a one-pot, two-step, three-component reaction (3-CR). In this design, two branched tails are appended to lipidoids containing two short body tails via degradable linkers, which dramatically boosts mRNA delivery transiently, akin to the behavior of solid rocket boosters. Two combinatorial libraries of SRB-lipidoids were systematically synthesized and screened by varying the headgroup structure, length of the body tails, and length of the branch tails. Multiple SRB-lipidoids were identified which afford potent LNPs, which were comparable to or more potent than the benchmark, approved DLin-MC3-DMA (MC3) LNP in vivo. Moreover, key structural criteria involving total carbon number, symmetry, and headgroup were identified, which could be used to faithfully predict the performance of unidentified SRB-lipidoids. Finally, the utility of a non-limiting, exemplary SRB-lipidoid of the present disclosure was demonstrated through hepatic delivery of human fibroblast growth factor 21 (FGF21)- encoded mRNA and co-delivery of Cas9 mRNA and sgRNA targeting transthyretin (TTR), respectively. With approximately 5-fold greater therapeutic FGF21 protein expression and TTR editing efficiency than MC3 LNP, the exemplary SRB-lipidoid, constructed via a practical synthetic method, demonstrates great promise for mRNA-based protein supplementation therapy and gene editing therapy. - 9 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 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 “adjuvant” as used herein is defined as any molecule to enhance an antigen- specific adaptive immune response. The term “alkenyl” as used herein refers to straight and branched chain and cyclic alkyl groups as defined herein, except that at least one double bond exists between two carbon atoms. Thus, alkenyl groups have from 2 to 40 carbon atoms, or 2 to about 20 carbon atoms, or 2 to 12 carbon atoms or, in some embodiments, from 2 to 8 carbon atoms. Examples include, but are not limited to vinyl, -CH=C=CCH2, -CH=CH(CH3), - CH=C(CH3)2, -C(CH3)=CH2, -C(CH3)=CH(CH3), -C(CH2CH3)=CH2, cyclohexenyl, cyclopentenyl, cyclohexadienyl, butadienyl, pentadienyl, and hexadienyl among others. The term “alkoxy” as used herein refers to an oxygen atom connected to an alkyl group, including a cycloalkyl group, as are defined herein. Examples of linear alkoxy groups include but are not limited to methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, and the like. Examples of branched alkoxy include but are not limited to isopropoxy, sec-butoxy, tert-butoxy, isopentyloxy, isohexyloxy, and the like. Examples of cyclic alkoxy include but are not limited to cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and the like. An alkoxy group can include about 1 to about 12, about 1 to about 20, or about 1 to about 40 carbon atoms bonded to the oxygen atom, and can further include double or triple bonds, and can also include heteroatoms. For example, an allyloxy group or a methoxyethoxy group is also an alkoxy group within the meaning herein, as is a methylenedioxy group in a context where two adjacent atoms of a structure are substituted therewith. The term “alkyl” as used herein refers to straight chain and branched alkyl groups and cycloalkyl groups having from 1 to 40 carbon atoms, 1 to about 20 carbon atoms, 1 to 12 carbons or, in some embodiments, from 1 to 8 carbon atoms. Examples of straight chain alkyl groups include those with from 1 to 8 carbon atoms such as methyl, ethyl, n-propyl, n-butyl, 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. - 10 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) The term “alkynyl” as used herein refers to straight and branched chain alkyl groups, except that at least one triple bond exists between two carbon atoms. Thus, alkynyl groups have from 2 to 40 carbon atoms, 2 to about 20 carbon atoms, or from 2 to 12 carbons or, in some embodiments, from 2 to 8 carbon atoms. Examples include, but are not limited to – C ^CH, -C ^C(CH3), -C ^C(CH2CH3), -CH2C ^CH, -CH2C ^C(CH3), and -CH2C ^C(CH2CH3) among others. The term “alkylene” or “alkylenyl” as used herein refers to a bivalent saturated aliphatic radical (e.g., -CH2-, -CH2CH2-, and -CH2CH2CH2-, inter alia). In certain embodiments, the term may be regarded as a moiety derived from an alkene by opening of the double bond or from an alkane by removal of two hydrogen atoms from the same (e.g., - CH2-) different (e.g., -CH2CH2-) carbon atoms. Similarly, the terms “heteroalkylenyl”, “cycloalkylenyl”, “heterocycloalkylenyl”, and the like, as used herein, refer to a divalent radical of the moiety corresponding to the base group (e.g., heteroalkyl, cycloalkyl, and / or heterocycloalkyl). A divalent radical possesses two open valencies at any position(s) of the group, wherein each radical may be on a carbon atom or heteroatom. Thus, the divalent radical may form a single bond to two distinct atoms or groups, or may form a double bond with one atom. The term “antigen” or “Ag” as used herein is defined as a molecule that provokes an adaptive immune response. This immune response may involve either antibody production, or the activation of specific immunogenically-competent cells, or both. The skilled artisan will understand that any macromolecule, including virtually all proteins or peptides, can serve as an antigen. Furthermore, antigens can be derived from recombinant or genomic DNA or RNA. A skilled artisan will understand that any DNA or RNA, which comprises a nucleotide sequences or a partial nucleotide sequence encoding a protein that elicits an adaptive immune response therefore encodes an “antigen” as that term is used herein. Furthermore, one skilled in the art will understand that an antigen need not be encoded solely by a full length nucleotide sequence of a gene. It is readily apparent that the present invention includes, but is 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. - 11 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) The term “amine” as used herein refers to primary, secondary, and tertiary amines having, e.g., the formula N(group)3wherein each group can independently be H or non-H, such as alkyl, aryl, and the like. Amines include but are not limited to R-NH2, for example, alkylamines, arylamines, alkylarylamines; R2NH wherein each R is independently selected, such as dialkylamines, diarylamines, aralkylamines, heterocyclylamines and the like; and R3N wherein each R is independently selected, such as trialkylamines, dialkylarylamines, alkyldiarylamines, triarylamines, and the like. The term “amine” also includes ammonium ions as used herein. The term “amino group” as used herein refers to a substituent of the form -NH2, - NHR, -NR2, -NR3+, wherein each R is independently selected, and protonated forms of each, except for -NR3+, which cannot be protonated. Accordingly, any compound substituted with an amino group can be viewed as an amine. An “amino group” within the meaning herein can be a primary, secondary, tertiary, or quaternary amino group. An “alkylamino” group includes a monoalkylamino, dialkylamino, and trialkylamino group. The term “anionic lipid” refers to any lipid that is negatively charged at physiological pH. These lipids include phosphatidylglycerol, cardiolipin, diacylphosphatidylserine, diacylphosphatidic acid, N-dodecanoylphosphatidylethanolamines, N- succinylphosphatidylethanolamines, N-glutarylphosphatidylethanolamines, lysylphosphatidylglycerols, palmitoyloleyolphosphatidylglycerol (POPG), and other anionic modifying groups joined to neutral lipids. The term “aryl” as used herein refers to cyclic aromatic hydrocarbon groups that do not contain heteroatoms in the ring. Thus aryl groups include, but are not limited to, phenyl, azulenyl, heptalenyl, biphenyl, indacenyl, fluorenyl, phenanthrenyl, triphenylenyl, pyrenyl, naphthacenyl, chrysenyl, biphenylenyl, anthracenyl, and naphthyl groups. In some embodiments, aryl groups contain about 6 to about 14 carbons in the ring portions of the groups. Aryl groups can be unsubstituted or substituted, as defined herein. Representative substituted aryl groups can be mono-substituted or substituted more than once, such as, but not limited to, a phenyl group substituted at any one or more of 2-, 3-, 4-, 5-, or 6-positions of 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 - 12 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) increased membrane fluidity. A number of cationic lipids and related analogs, which are also useful in the present disclosure, have been described in U.S. Patent Publication Nos. 20060083780 and 20060240554; U.S. Patent Nos.5,208,036; 5,264,618; 5,279,833; 5,283,185; 5,753,613; and 5,785,992; and PCT Publication No. WO 96 / 10390, the disclosures of which are herein incorporated by reference in their entirety for all purposes. Non-limiting examples of cationic lipids are described in detail herein. In some cases, the cat-ionic lipids comprise a protonatable tertiary amine (e.g., pH titratable) head group, C18 alkyl chains, ether linkages between the head group and alkyl chains, and 0 to 3 double bonds. Such lipids include, e.g., DSDMA, DLinDMA, DLenDMA, and DODMA. The term “cycloalkyl” as used herein refers to cyclic alkyl groups such as, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl groups. In some embodiments, the cycloalkyl group can have 3 to about 8-12 ring members, whereas in other embodiments the number of ring carbon atoms range from 3 to 4, 5, 6, or 7. Cycloalkyl groups further include polycyclic cycloalkyl groups such as, but not limited to, norbornyl, adamantyl, bornyl, camphenyl, isocamphenyl, and carenyl groups, and fused rings such as, but not limited to, decalinyl, and the like. Cycloalkyl groups also include rings that are substituted with straight or branched chain alkyl groups as defined herein. Representative substituted cycloalkyl groups can be mono-substituted or substituted more than once, such as, but not limited to, 2,2-, 2,3-, 2,4- 2,5- or 2,6-disubstituted cyclohexyl groups or mono-, di- or tri-substituted norbornyl or cycloheptyl groups, which can be substituted with, for example, amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups. The term “cycloalkenyl” alone or in combination denotes a cyclic alkenyl group. A “disease” is a state of health of an animal wherein the animal cannot maintain homeostasis, and wherein if the disease is not ameliorated then the animal’s health continues to deteriorate. In contrast, a “disorder” in an animal is a state of health in which the animal is able to maintain homeostasis, but in which the animal’s state of health is less favorable than it would be in the absence of the disorder. Left untreated, a disorder does not necessarily cause a 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 - 13 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) to provide the desired biological result. That result may be reduction and / or alleviation of the signs, symptoms, or causes of a disease, or any other desired alteration of a biological system. An appropriate therapeutic amount in any individual case may be determined by one of ordinary skill in the art using routine experimentation. In particular, in the case of a mRNA, and “effective amount” or “therapeutically effective amount” of a therapeutic nucleic acid as relating to a mRNA is an amount sufficient to produce the desired effect, e.g., mRNA-directed expression of an amount of a protein that causes a desirable biological effect in the organism within which the protein is expressed. For example, in some embodiments, the expressed protein is an active form of a protein that is normally expressed in a cell type within the body, and the therapeutically effective amount of the mRNA is an amount that produces an amount of the encoded protein that is at least 50% (e.g., at least 60%, or at least 70%, or at least 80%, or at least 90%) of the amount of the protein that is normally expressed in the cell type of a healthy individual. For example, in some embodiments, the expressed protein is a protein that is normally expressed in a cell type within the body, and the therapeutically effective amount of the mRNA is an amount that produces a similar level of expression as observed in a healthy individual in an individual with aberrant expression of the protein (i.e., protein deficient individual). Suitable assays for measuring the expression of an mRNA or protein include, but are not limited to dot blots, Northern blots, in situ hybridization, ELISA, immunoprecipitation, enzyme function, as well as phenotypic assays known to those of skill in the art. The term “encode” as used herein refers to the product specified (e.g., protein and RNA) by a given sequence of nucleotides in a nucleic acid (i.e., DNA and / or RNA), upon transcription or translation of the DNA or RNA, respectively. In certain embodiments, the term “encode” refers to the RNA sequence specified by transcription of a DNA sequence. In certain embodiments, the term “encode” refers to the amino acid sequence (e.g., polypeptide or protein) specified by translation of mRNA. In certain embodiments, the term “encode” refers to the amino acid sequence specified by transcription of DNA to mRNA and subsequent translation of the mRNA encoded by the DNA sequence. In certain embodiments, 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 - 14 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) encapsulated system, preferably less than about 25% of the active agent or therapeutic agent in the particle is degraded in a treatment that would normally degrade 100% of free active agent or therapeutic agent, more preferably less than about 10%, and most preferably less than about 5% of the active agent or therapeutic agent in the particle is degraded. In the context of nucleic acid therapeutic agents, full encapsulation may be determined by an OLIGREEN® assay. OLIGREEN® is an ultra-sensitive fluorescent nucleic acid stain for quantitating oligonucleotides and single-stranded DNA in solution (available from Invitrogen Corporation; Carlsbad, Calif.). RiboGreen is an ultra-sensitive fluorescent nucleic acid stain for quantitating RNA in solution. “Fully encapsulated” also indicates that the lipid particles are serum stable, that is, that they do not rapidly decompose into their component parts upon in vivo administration. The terms “halo,” “halogen,” or “halide” group, as used herein, by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom. The term “haloalkyl” group, as used herein, includes mono-halo alkyl groups, poly- halo alkyl groups wherein all halo atoms can be the same or different, and per-halo alkyl groups, wherein all hydrogen atoms are replaced by halogen atoms, such as fluoro. Examples of haloalkyl include trifluoromethyl, 1,1-dichloroethyl, 1,2-dichloroethyl, 1,3-dibromo-3,3- difluoropropyl, perfluorobutyl, and the like. The term “helper lipid” as used herein refers to a lipid capable of increasing the effectiveness of delivery of lipid-based particles such as cationic lipid-based particles to a target, preferably into a cell. The helper lipid can be neutral, positively charged, or negatively charged. In certain embodiments, the helper lipid is neutral or negatively charged. Non- limiting examples of helper lipids include 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphoethanolamine (DOPE), 1-palmitoyl- 2-oleoyl-sn-glycero-3phosphocholin (POPC) and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC). The term “heteroalkyl” as used herein by itself or in combination with another term, 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 - 15 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) molecule. The term “heteroaryl” as used herein refers to aromatic ring compounds containing 5 or more ring members, of which, one or more is a heteroatom such as, but not limited to, N, O, and S; for instance, heteroaryl rings can have 5 to about 8-12 ring members. A heteroaryl group is a variety of a heterocyclyl group that possesses an aromatic electronic structure. A heteroaryl group designated as a C2-heteroaryl can be a 5-ring with two carbon atoms and three heteroatoms, a 6-ring with two carbon atoms and four heteroatoms and so forth. Likewise a C4-heteroaryl can be a 5-ring with one heteroatom, a 6-ring with two heteroatoms, and so forth. The number of carbon atoms plus the number of heteroatoms sums up to equal the total number of ring atoms. Heteroaryl groups include, but are not limited to, groups such as pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridinyl, thiophenyl, benzothiophenyl, benzofuranyl, indolyl, azaindolyl, indazolyl, benzimidazolyl, azabenzimidazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, imidazopyridinyl, isoxazolopyridinyl, thianaphthalenyl, purinyl, xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, quinoxalinyl, and quinazolinyl groups. Heteroaryl groups can be unsubstituted, or can be substituted with groups as is discussed herein. Representative substituted heteroaryl groups can be substituted one or more times with groups such as those listed herein. Additional examples of aryl and heteroaryl groups include but are not limited to phenyl, biphenyl, indenyl, naphthyl (1-naphthyl, 2-naphthyl), N-hydroxytetrazolyl, N- hydroxytriazolyl, N-hydroxyimidazolyl, anthracenyl (1-anthracenyl, 2-anthracenyl, 3- anthracenyl), thiophenyl (2-thienyl, 3-thienyl), furyl (2-furyl, 3-furyl) , indolyl, oxadiazolyl, isoxazolyl, quinazolinyl, fluorenyl, xanthenyl, isoindanyl, benzhydryl, acridinyl, thiazolyl, pyrrolyl (2-pyrrolyl), pyrazolyl (3-pyrazolyl), imidazolyl (1-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl), triazolyl (1,2,3-triazol-1-yl, 1,2,3-triazol-2-yl 1,2,3-triazol-4-yl, 1,2,4-triazol-3-yl), oxazolyl (2-oxazolyl, 4-oxazolyl, 5-oxazolyl), thiazolyl (2-thiazolyl, 4- thiazolyl, 5-thiazolyl), pyridyl (2-pyridyl, 3-pyridyl, 4-pyridyl), pyrimidinyl (2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, 6-pyrimidinyl), pyrazinyl, pyridazinyl (3- pyridazinyl, 4- 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), - 16 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 6-(2,3-dihydro-benzo[b]furanyl), 7-(2,3-dihydro-benzo[b]furanyl), benzo[b]thiophenyl (2- benzo[b]thiophenyl, 3-benzo[b]thiophenyl, 4-benzo[b]thiophenyl, 5-benzo[b]thiophenyl, 6- benzo[b]thiophenyl, 7-benzo[b]thiophenyl), 2,3-dihydro-benzo[b]thiophenyl, (2-(2,3- dihydro-benzo[b]thiophenyl), 3-(2,3-dihydro-benzo[b]thiophenyl), 4-(2,3-dihydro- benzo[b]thiophenyl), 5-(2,3-dihydro-benzo[b]thiophenyl), 6-(2,3-dihydro- benzo[b]thiophenyl), 7-(2,3-dihydro-benzo[b]thiophenyl), indolyl (1-indolyl, 2-indolyl, 3-indolyl, 4-indolyl, 5-indolyl, 6-indolyl, 7-indolyl), indazole (1-indazolyl, 3-indazolyl, 4-indazolyl, 5-indazolyl, 6-indazolyl, 7-indazolyl), benzimidazolyl (1-benzimidazolyl, 2-benzimidazolyl, 4-benzimidazolyl, 5-benzimidazolyl, 6-benzimidazolyl, 7-benzimidazolyl, 8-benzimidazolyl), benzoxazolyl (1-benzoxazolyl, 2-benzoxazolyl), benzothiazolyl (1- benzothiazolyl, 2-benzothiazolyl, 4-benzothiazolyl, 5-benzothiazolyl, 6-benzothiazolyl, 7-benzothiazolyl), carbazolyl (1-carbazolyl, 2-carbazolyl, 3-carbazolyl, 4-carbazolyl), 5H-dibenz[b,f]azepine (5H-dibenz[b,f]azepin-1-yl, 5H-dibenz[b,f]azepine-2-yl, 5H-dibenz[b,f]azepine-3-yl, 5H-dibenz[b,f]azepine-4-yl, 5H-dibenz[b,f]azepine-5-yl), 10,11-dihydro-5H-dibenz[b,f]azepine (10,11-dihydro-5H-dibenz[b,f]azepine-1-yl, 10,11-dihydro-5H-dibenz[b,f]azepine-2-yl, 10,11-dihydro-5H-dibenz[b,f]azepine-3-yl, 10,11-dihydro-5H-dibenz[b,f]azepine-4-yl, 10,11-dihydro-5H-dibenz[b,f]azepine-5-yl), and the like. The term “heterocycloalkyl” as used herein refers to an aliphatic, partially unsaturated or fully saturated, 3- to 14-membered ring system, including single rings of 3 to 8 atoms and bi- and tricyclic ring systems where at least one of the carbon atoms of the ring is replaced with a heteroatom such as, but not limited to, nitrogen, oxygen, sulfur, or phosphorus. A heterocycloalkyl can include one to four heteroatoms independently selected from oxygen, nitrogen, and sulfur, wherein a nitrogen and sulfur heteroatom optionally can be oxidized and a nitrogen heteroatom optionally can be substituted. Representative heterocycloalkyl groups include, but are not limited, to the following exemplary groups: pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, and tetrahydrofuryl. 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 - 17 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) carbon atoms and three heteroatoms, a 6-ring with two carbon atoms and four heteroatoms and so forth. Likewise a C4-heterocyclyl can be a 5-ring with one heteroatom, a 6-ring with two heteroatoms, and so forth. The number of carbon atoms plus the number of heteroatoms equals the total number of ring atoms. A heterocyclyl ring can also include one or more double bonds. A heteroaryl ring is an embodiment of a heterocyclyl group. The phrase “heterocyclyl group” includes fused ring species including those that include fused aromatic and non-aromatic groups. For example, a dioxolanyl ring and a benzdioxolanyl ring system (methylenedioxyphenyl ring system) are both heterocyclyl groups within the meaning herein. The phrase also includes polycyclic ring systems containing a heteroatom such as, but not limited to, quinuclidyl. Heterocyclyl groups can be unsubstituted, or can be substituted as discussed herein. Heterocyclyl groups include, but are not limited to, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridinyl, thiophenyl, benzothiophenyl, benzofuranyl, dihydrobenzofuranyl, indolyl, dihydroindolyl, azaindolyl, indazolyl, benzimidazolyl, azabenzimidazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, imidazopyridinyl, isoxazolopyridinyl, thianaphthalenyl, purinyl, xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, quinoxalinyl, and quinazolinyl groups. Representative substituted heterocyclyl groups can be mono-substituted or substituted more than once, such as, but not limited to, piperidinyl or quinolinyl groups, which are 2-, 3-, 4-, 5-, or 6- substituted, or disubstituted with groups such as those listed herein. The term “hydrocarbon” or “hydrocarbyl” as used herein refers to a molecule or functional group that includes carbon and hydrogen atoms. The term can also refer to a molecule or functional group that normally includes both carbon and hydrogen atoms but wherein all the hydrogen atoms are substituted with other functional groups. As used herein, the term “hydrocarbyl” refers to a functional group derived from a straight chain, branched, or cyclic hydrocarbon, and can be alkyl, alkenyl, alkynyl, aryl, cycloalkyl, acyl, or any combination thereof. Hydrocarbyl groups can be shown as (Ca- Cb)hydrocarbyl, wherein a and b are integers and mean having any of a to b number of carbon 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 “immune cell,” as used herein refers to any cell involved in the mounting of an immune response. Such cells include, but are not limited to, T cells, B cells, NK cells, antigen-presenting cells (e.g., dendritic cells and macrophages), monocytes, neutrophils, - 18 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) eosinophils, basophils, and the like. The term “independently selected from” as used herein refers to referenced groups being the same, different, or a mixture thereof, unless the context clearly indicates otherwise. Thus, under this definition, the phrase “X1, X2, and X3are independently selected from noble gases” would include the scenario where, for example, X1, X2, and X3are all the same, where X1, X2, and X3are all different, where X1and X2are the same but X3is different, and other analogous permutations. The term “ionizable lipid” as used herein refers to a lipid (e.g., a cationic lipid) or lipidoid having at least one protonatable or deprotonatable group, such that the lipid is positively charged at a pH at or below physiological pH (e.g., pH 7.4), and neutral at a second pH, preferably at or above physiological pH. It will be understood by one of ordinary skill in the art that the addition or removal of protons as a function of pH is an equilibrium process, and that the reference to a charged or neutral lipid refers to the nature of the predominant species and does not require that all of the lipid be present in the charged or neutral form. Generally, ionizable lipids have a pKa of the protonatable group in the range of about 4 to about 7. The term “local delivery,” as used herein, refers to delivery of an active agent or therapeutic agent such as a messenger RNA directly to a target site within an organism. For example, an agent can be locally delivered by direct injection into a disease site such as a tumor or other target site such as a site of inflammation or a target organ such as the liver, heart, pancreas, kidney, and the like. The term “lipid” refers to a group of organic compounds that include, but are not limited to, esters of fatty acids and are characterized by being insoluble in water, but soluble in many organic solvents. They are usually divided into at least three classes: (1) “simple lipids,” which include fats and oils as well as waxes; (2) “compound lipids,” which include phospholipids and glycolipids; and (3) “derived lipids” such as steroids. 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 - 19 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 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. 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 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” 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. - 20 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 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. Biol. Chem., 260:2605-2608 (1985); Rossolini et al., Mal. Cell. Probes, 8:91-98 (1994)). As used herein, the term “nucleic acid” includes any oligonucleotide or polynucleotide, with fragments containing up to 60 nucleotides generally termed oligonucleotides, and longer fragments termed polynucleotides. In particular embodiments, oligonucleotides of the disclosure are from about 15 to about 60 nucleotides in length. Nucleic acid may be administered alone in the lipid particles of the disclosure, or in combination (e.g., co-administered) with lipid particles of the disclosure comprising peptides, polypeptides, or small molecules such as conventional drugs. In other embodiments, the nucleic acid may be administered in a viral vector. “Nucleotides” contain a sugar deoxyribose (DNA) or ribose (RNA), a base, and a phosphate group. Nucleotides are linked together through the phosphate groups. “Bases” - 21 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 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 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, - 22 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 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 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 - 23 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 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 term “polymer conjugated lipid” refers to a molecule comprising both a lipid portion and a polymer portion. An example of a polymer conjugated lipid is a pegylated lipid. The term “pegylated lipid” refers to a molecule comprising both a lipid portion and a polyethylene glycol portion. Pegylated lipids are known in the art and include 1-(monomethoxy-polyethyleneglycol)-2,3-dimyristoylglycerol (PEG-s- DMG), DSPE-PEG- DBCO, DOPE-PEG-Azide, DSPE-PEG-Azide, DPPE-PEG-Azide, DSPE-PEG-Carboxy- NHS, DOPE-PEG-Carboxylic Acid, DSPE-PEG-Carboxylic acid and the like. The term “room temperature” as used herein refers to a temperature of about 15 °C to 28 °C. The term “solvent” as used herein refers to a liquid that can dissolve a solid, liquid, or gas. Non-limiting examples of solvents are silicones, organic compounds, water, alcohols, ionic liquids, and supercritical fluids. 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 - 24 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 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, 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 “substituted” as used herein in conjunction with a molecule or an organic group as defined herein refers to the state in which one or more hydrogen atoms contained therein are replaced by one or more non-hydrogen atoms. The term “functional group” or “substituent” as used herein refers to a group that can be or is substituted onto a molecule or onto an organic group. Examples of substituents or functional groups include, but are not limited to, a halogen (e.g., F, Cl, Br, and I); an oxygen atom in groups such as hydroxy groups, alkoxy groups, aryloxy groups, aralkyloxy groups, oxo(carbonyl) groups, carboxyl groups including carboxylic acids, carboxylates, and carboxylate esters; a sulfur atom in groups such as thiol groups, alkyl and aryl sulfide groups, sulfoxide groups, sulfone groups, sulfonyl groups, and sulfonamide groups; a nitrogen atom in groups such as amines, - 25 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 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 protein” as used herein refers to a protein or peptide which has a positive or advantageous effect on a condition or disease state of a subject when provided to the subject in a therapeutically effective amount. In some embodiments, a therapeutic protein or peptide has curative or palliative properties and may be administered to ameliorate, relieve, alleviate, reverse, delay onset of or lessen the severity of one or more symptoms of a disease or disorder. A therapeutic protein or peptide may have prophylactic properties and may be used to delay the onset of a disease or to lessen the severity of such disease or pathological condition. The term “therapeutic protein” includes entire proteins or peptides, and can also refer to therapeutically active fragments thereof. It can also include therapeutically active variants of a protein. Exemplary therapeutic proteins include, but are not limited to, an analgesic protein, an anti-inflammatory protein, an anti-proliferative protein, an proapoptotic protein, an anti-angiogenic protein, a cytotoxic protein, a cytostatic protein, a cytokine, a chemokine, a growth factor, a wound healing protein, a pharmaceutical protein, or a pro-drug activating protein. Therapeutic proteins may include growth factors (EGF, TGF-α, TGF- β, TNF, HGF, IGF, and IL-1-8, inter alia) cytokines, paratopes, Fabs (fragments, antigen binding), and antibodies. The terms “treat,” “treating” and “treatment,” as used herein, means reducing the frequency or severity with which symptoms of a disease or condition are experienced by a subject by virtue of administering an agent or compound to the subject. - 26 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) Ionizable Lipid (Lipidoid) Compounds In one aspect, the present disclosure provides a compound formula (I), or a salt or solvate thereof: (I), wherein: A is selected from the group consisting ; R1aand R1bare each ; R2is selected from the group substituted C1-C6alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C6 heteroalkyl, optionally substituted C3-C6 heteroalkenyl, optionally substituted C2-C8heterocycloalkyl, and N(Ra)(Rb); R3aand R3bare each independently selected from the group consisting of optionally substituted C1-C6alkyl, optionally substituted C2-C6alkenyl, optionally substituted C3-C8cycloalkyl, optionally substituted C2-C6 heteroalkyl, optionally substituted C3-C6 heteroalkenyl, and optionally substituted C2-C8heterocycloalkyl; each occurrence of R4aand R4bis independently H; each occurrence of R5ais independently selected from the group consisting of optionally substituted C1-C24 alkyl, optionally substituted C3-C12 cycloalkyl, optionally substituted C2-C24heteroalkyl, optionally substituted C2-C12heterocycloalkyl, optionally substituted C2-C24 alkylenyl, optionally substituted C3-C24 heteroalkylenyl, optionally substituted C2-C24alkynylenyl, and optionally substituted C3-C24heteroalkynylenyl; each occurrence of R5bis H; each occurrence of R6is independently selected from the group consisting of optionally substituted C1-C24 alkyl, optionally substituted C3-C12 cycloalkyl, optionally substituted C2-C24heteroalkyl, optionally substituted C2-C12heterocycloalkyl, optionally substituted C2-C24 alkylenyl, optionally substituted C3-C24 heteroalkylenyl, optionally substituted C2-C24alkynylenyl, and optionally substituted C3-C24heteroalkynylenyl; L is selected from the group consisting of optionally substituted C2-C6 alkylenyl and - 27 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) optionally substituted C2-C8 heteroalkylenyl; and Raand Rbare each independently selected from the group consisting of H and optionally substituted C1-C6 alkyl. In certain embodiments, the compound of formula (I) is a compound of formula (Ia): (Ia). In certain embodiments, the compound of formula (I) is a compound of formula (Ia- 1): . In certain a compound of formula (Ia- 2): (Ia-2). In certain embodiments, the compound of formula (I) is a compound of formula (Ia- 3): (Ia-3). In certain embodiments, the compound of formula (I) is a compound of formula (Ia- 4): (Ia-4). In certain embodiments, the compound of formula (I) is a compound of formula (Ib): . In certain is a compound of formula (Ib- 1): . In certain is a compound of formula (Ib- 2): - 28 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) . In certain is a compound of formula (Ib- 3): . In certain is a compound of formula (Ib- 4): . In certain substituted alkyl, optionally substituted alkenyl, substituted cycloalkyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted alkynylenyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted alkylenyl, and optionally substituted heteroalkylenyl 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-C3haloalkoxy, 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-C8heteroaryl, 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-C6alkyl, C3-C8cycloalkyl, C1-C6haloalkyl, benzyl, and phenyl, or wherein R’ and R’’ can combine with the nitrogen atom to which they are bound to form a C2-C8heterocycloalkyl (e.g., cyclic tertiary amine). In certain embodiments, each occurrence of R5ais different. In certain embodiments, each occurrence of R5ais identical. In certain embodiments, R5ais ethyl. In certain embodiments, R5ais propyl. In certain embodiments, R5ais butyl. In certain embodiments, R5ais pentyl. In certain embodiments, R5ais hexyl. In certain embodiments, R5ais heptyl. In certain embodiments, R5ais octyl. In certain embodiments, R5ais nonyl. In certain embodiments, R5ais decyl. In certain embodiments, R5ais undecyl. In certain embodiments, R5ais dodecyl. In certain embodiments, R5ais tridecyl. In certain embodiments, R5ais tetradecyl. In certain embodiments, R5ais n-propyl. In certain embodiments, R5ais n-pentyl. In certain embodiments, R5ais n-heptyl. In certain embodiments, R5ais n-nonyl. In certain - 29 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) embodiments, R5ais n-undecyl. In certain embodiments, R5ais n-tridecyl. In certain embodiments, each occurrence of R6is different. In certain embodiments, each occurrence of R6is identical. In certain embodiments, R6is ethyl. In certain embodiments, R6is propyl. In certain embodiments, R6is butyl. In certain embodiments, R6is pentyl. In certain embodiments, R6is hexyl. In certain embodiments, R6is heptyl. In certain embodiments, R6is octyl. In certain embodiments, R6is nonyl. In certain embodiments, R6is decyl. In certain embodiments, R6is undecyl. In certain embodiments, R6is dodecyl. In certain embodiments, R6is tridecyl. tetradecyl. In certain embodiments, R6is n-butyl. In certain embodiments, R6is n-hexyl. In certain embodiments, R6is n-octyl. In certain embodiments, R6is n-decyl. In certain embodiments, R6is n-dodecyl. In certain embodiments, R1aand R1bare identical. In certain embodiments, R1aand R1bare different. In certain embodiments, R1a. In certain embodiments, R1ais . In certain embodiments, R1a. In certain embodiments, R1ais embodiments, R1a. In certain embodiments, R1a. In certain embodiments, R1a. In certain embodiments, R1a. In certain embodiments, R1a. In certain embodiments, R1a. In certain embodiments, R1a. In certain embodiments, R1a. In certain embodiments, R1a. In certain embodiments, R1a. In certain embodiments, R1a. In certain embodiments, R1a. In certain - 30 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) embodiments, R1ais . In certain embodiments, R1a, . In certain embodiments, R1a. In certain embodiments, R1ais . In certain embodiments, R1a. In certain embodiments, R1ais . In certain embodiments, R1b. In certain embodiments, R1bis . In certain embodiments, R1b. In certain embodiments, R1bis . In certain embodiments, R1b. In certain embodiments, R1b. In certain embodiments, R1b. In certain embodiments, R1b. In certain embodiments, R1b. In certain embodiments, R1b. In certain embodiments, R1b. In certain embodiments, R1b. In certain embodiments, R1b. In certain embodiments, R1b. In certain embodiments, R1b. In certain embodiments, R1b. In certain embodiments, R1b. In certain embodiments, R1b. In certain embodiments, R1b, . In certain embodiments, R1b. In certain embodiments, R1bis - 31 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) . In certain embodiments, R1bis . In certain embodiments, R1bis . embodiments, R5aand R6are selected such that R1aand R1beach comprise about 14, 15, 16, 17, 18, 19, 20, 21, or 22 carbon atoms. In certain embodiments, R5aand R6are selected such that R1aand R1beach independently comprise about 18 carbon atoms. In certain embodiments, R2is selected from the group consisting of C2-C3alkyl substituted with at least one cyclic tertiary amine or acyclic tertiary amine. In certain embodiments, R2is a C2-C6alkyl substituted with at least one tertiary amine. In certain embodiments, R2is a C2 alkyl substituted with at least one tertiary amine. In certain embodiments, R2is a C3alkyl substituted with at least one tertiary amine. In certain embodiments, the tertiary amine is a acyclic tertiary amine (e.g., dimethylamine, diethylamine, dipropylamine, methylethylamine, and the like). In certain embodiments, the tertiary amine is a cyclic tertiary amine (e.g., azetidine, pyrrolidine, piperidine, piperazine, imidazole, pyridine, and triazole, and the like). In certain embodiments, R2is Me. In certain embodiments, R2is Et. In certain embodiments, R2is N(Me)2. In certain embodiments, R2. In certain embodiments, R2is . In certain embodiments, R2. In certain embodiments, R2. In certain embodiments, R2. In certain embodiments, R2. In certain embodiments, R2. In certain embodiments, R2 . In certain embodiments, R2. In certain embodiments, R2. In certain embodiments, R2is . L is -(CH2)3-. In - 32 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) certain embodiments, L is -(CH2)3N(Me)(CH2)3-. In certain embodiments, R3aand R3bare identical. In certain embodiments, R3aand R3bare different. In certain embodiments, R3ais Me. In certain embodiments, R3ais Et. In certain embodiments, R3bis Me. In certain embodiments, R3bis Et. In certain embodiments, R3aand R3bcombine with the nitrogen atoms to which they are bound to form . In certain embodiments, R3aand R3bcombine with the nitrogen atoms to which they are bound to form . In certain embodiments, A is . In certain embodiments, A is . In certain embodiments, A . In certain embodiments, A . In certain embodiments, A . In certain embodiments, A . In certain embodiments, A . In certain embodiments, A . In certain embodiments, A is . In certain embodiments, A . In certain embodiments, . In certain embodiments, A . In certain In certain embodiments, A is . In certain embodiments, A . In - 33 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) certain embodiments, A is . In certain from the group consisting of: ((3- 1,2-diyl) dihexanoate; ((3-(dimethylamino)propyl)azanediyl)bis(hexane-1,2-diyl) dioctanoate; ((3-(dimethylamino)propyl)azanediyl)bis(hexane-1,2-diyl) bis(decanoate); ((3-(dimethylamino)propyl)azanediyl)bis(hexane-1,2-diyl) didodecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(hexane-1,2-diyl) ditetradecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(octane-1,2-diyl) dihexanoate; ((3-(dimethylamino)propyl)azanediyl)bis(octane-1,2-diyl) dioctanoate; ((3-(dimethylamino)propyl)azanediyl)bis(octane-1,2-diyl) bis(decanoate); ((3-(dimethylamino)propyl)azanediyl)bis(octane-1,2-diyl) didodecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(octane-1,2-diyl) ditetradecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(decane-1,2-diyl) dihexanoate; ((3-(dimethylamino)propyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((3-(dimethylamino)propyl)azanediyl)bis(decane-1,2-diyl) bis(decanoate); ((3-(dimethylamino)propyl)azanediyl)bis(decane-1,2-diyl) didodecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(decane-1,2-diyl) ditetradecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(dodecane-1,2-diyl) dihexanoate; ((3-(dimethylamino)propyl)azanediyl)bis(dodecane-1,2-diyl) dioctanoate; ((3-(dimethylamino)propyl)azanediyl)bis(dodecane-1,2-diyl) bis(decanoate); ((3-(dimethylamino)propyl)azanediyl)bis(dodecane-1,2-diyl) didodecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(dodecane-1,2-diyl) ditetradecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(tetradecane-1,2-diyl) dihexanoate; ((3-(dimethylamino)propyl)azanediyl)bis(tetradecane-1,2-diyl) dioctanoate; ((3-(dimethylamino)propyl)azanediyl)bis(tetradecane-1,2-diyl) bis(decanoate); ((3-(dimethylamino)propyl)azanediyl)bis(tetradecane-1,2-diyl) didodecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(tetradecane-1,2-diyl) ditetradecanoate; (methylazanediyl)bis(decane-1,2-diyl) dioctanoate; (ethylazanediyl)bis(decane-1,2-diyl) dioctanoate; (2,2-dimethylhydrazine-1,1-diyl)bis(decane-1,2-diyl) dioctanoate; ((4-methylpiperazin-1-yl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((2-methoxyethyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((2-(dimethylamino)ethyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; - 34 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) ((4-(dimethylamino)butyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((2-(diethylamino)ethyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((3-(diethylamino)propyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((2-(pyrrolidin-1-yl)ethyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((3-(pyrrolidin-1-yl)propyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((3-(1H-imidazol-1-yl)propyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((2-(piperidin-1-yl)ethyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((3-(4-methylpiperazin-1-yl)propyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; (ethane-1,2-diylbis(methylazanediyl))bis(decane-1,2-diyl) dioctanoate; (propane-1,3-diylbis(methylazanediyl))bis(decane-1,2-diyl) dioctanoate; (ethane-1,2-diylbis(ethylazanediyl))bis(decane-1,2-diyl) dioctanoate; (((methylazanediyl)bis(propane-3,1-diyl))bis(methylazanediyl))bis(decane-1,2-diyl) dioctanoate; and piperazine-1,4-diylbis(decane-1,2-diyl) dioctanoate. Ionizable Lipids and / or Cationic Lipids or Lipidoids The scope of ionizable lipids contemplated for use in the present disclosure is not limited to ionizable lipidoids of formula (I). 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 - 35 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) (DLinDAP), 1,2-dilinoleylthio-3-dimethylaminopropane (DLin-2-DMAP), 1,2-dilinoleyloxy- 3-trimethylaminopropane chloride salt (DLin-TMA.Cl), 1,2-dilinoleoyl-3- trimethylaminopropane chloride salt (DLin-TAP.Cl), 1,2-dilinoleyloxy-3-(N- methylpiperazino)propane (D Lin-MPZ), 3-(N,N-dilinoleylamino)-1,2-propanediol (D LinAP), 3-(N,N-dioleylamino)-1,2-propanedio (DOAP), 1,2-dilinoleyloxo-3-(2-N,N- dimethylamino)ethoxypropane (D Lin-EG-D MA), N,N-dioleyl-N,N-dimethylanrmonium chloride (DODAC), 1,2-dioleyloxy-N,N-dimethylaminopropane (DODMA), 1,2- distearyloxy-N,N-dimethylaminopropane (DSD MA), N-(1-(2,3-dioleyloxy)propyl)-N,N,N- trimethylammonium chloride (DOTMA), N,N-distearyl-N,N-dimethylammonium bromide (DDAB), N-(1-(2,3-dioleoyloxy)propyl)-N,N, N-trimethylammonium chloride (DOTAP), 3- (N-(N’,N’dimethylaminoethane)-carbamoyl)cholesterol (DC-Chol), N-(l,2- dimyristyloxyprop-3-yl)-N,N-dimethyl-N-hydroxyethyl anrmonium bromide (DMRIE), 2,3- dioleyloxy-N-[2 (spermine-carboxamidoethyl]-N,N-dimethy 1-1- propanaminiumtrifluoroacetate (DOSPA), dioctadecylamidoglycyl spermine (DOGS), 3- dimethylamino-2-(cholest-5-en-3-beta-oxybutan-4-oxy)-1-(cis,cis-9,12- octadecadienoxy)propane (CLinDMA), 2-[5’-(cholest-5-en-3-beta-oxy)-3’-oxapentoxy)-3- dimethyl-1-(cis,cis-9’,1-2’-octadecadienoxy) propane (CpLinDMA), N,N-dimethyl-3,4- dioleyloxybenzylamine (DMOBA), 1,2-N,N’dioleylcarbamyl-3-dimethylaminopropane (DOcarbDAP), 1,2-N,N’-dilinoleylcarbamyl-3-dimethylaminopropane (DLincarbDAP), or mixtures thereof. In certain embodiments, the cationic lipid is DLinDMA, DLin-K-C2-DMA (“XTC2”), or mixtures thereof. The ionizable lipids are not limited to those recited herein, and can further include ionizable lipids known to those skilled in the art, or described in PCT Application No. PCT / US2020 / 056255 and / or PCT Application No. PCT / US2020 / 056252, the disclosures of which are herein incorporated by reference in its entirety. The synthesis of cationic lipids such as DLin-K-C2-DMA (“XTC2”), DLin-K-C3- DMA, DLin-K-C4-DMA, DLin-K6-DMA, and DLin-K-MPZ, as well as additional cationic lipids, is described in U.S. Application Publication No. US 2011 / 0256175, the disclosure of which is herein incorporated by reference in its entirety for all purposes. The synthesis of cationic lipids such as DLin-K-DMA, DLin-CDAP, DLin-DAC, DLin-MA, DLinDAP, DLin-S-DMA, DLin-2-DMAP, DLin-TMA.Cl, DLin-TAP.Cl, DLin-MPZ, DLinAP, DOAP, 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. - 36 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 20060240554, the disclosure of which is herein incorporated by reference in its entirety for all purposes. Non-cationic Lipid In the nucleic acid-lipid particles of the present disclosure, the non-cationic lipid may comprise, e.g., one or more anionic lipids and / or neutral lipids. In some embodiments, the non-cationic lipid comprises one of the following neutral lipid components: (1) cholesterol or a derivative thereof (2) a phospholipid; or (3) a mixture of a phospholipid and cholesterol or a derivative thereof. Examples of cholesterol derivatives include, but are not limited to, cholestanol, cholestanone, cholestenone, coprostanol, cholesteryl-2’-hydroxyethyl ether, cholesteryl-4’- hydroxybutyl ether, and mixtures thereof. The synthesis of cholesteryl-2’-hydroxyethyl ether is known to one skilled in the art and described in U.S. Patent Nos.8,058,069, 8,492,359, 8,822,668, 9,364,435, 9,504,651, and 11,141,378, all of which are hereby incorporated herein in their entireties for all purposes. Non-limiting examples of non-cationic lipids include phospholipids such as lecithin, phosphatidylethanolamine, lysolecithin, lysophosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, egg sphingomyelin (ESM), cephalin, cardiolipin, phosphatidic acid, cerebrosides, dicetylphosphate, distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), ioleoylphosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoylphosphatidylethanolamine (POPE), palmitoyloleyolphosphatidylglycerol (POPG), dioleoylphosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-l- carboxylate DOPE-mal), dipalmitoylphosphatidylethanolamine (DPPE), dimyristoylphosphatidylethanolamine (DMPE), distearoylphosphatidylethanolamine (DSPE), monomethylphosphatidylethanolamine, dimethylphosphatidylethanolamine, dielaidoylphosphatidylethanolamine (DEPE), stearoyloleoylphosphatidylethanolamine (SOPE), lysophosphatidylcholine, dilinoleoylphosphatidylcholine, and mixtures thereof. Other diacylphosphatidylcholine and diacylphosphatidylethanolamine phospholipids 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- - 37 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 2’-hydroxyethyl ether, cholesteryl-4’-hydroxybutyl ether, and mixtures thereof. In certain embodiments, the phospholipid is DPPC, DSPC, or mixtures thereof. Conjugated Lipid In the nucleic acid-lipid particles of the present disclosure, the conjugated lipid that inhibits aggregation of particles may comprise, e.g., one or more of the following: a polyethyleneglycol (PEG) lipid conjugate, a polyamide (ATTA)-lipid conjugate, a cationic- polymer-lipid conjugates (CPLs), or mixtures thereof. In some embodiments, the nucleic acid-lipid particles comprise either a PEG-lipid conjugate or an ATTA-lipid conjugate. PEG is a linear, water-soluble polymer of ethylene PEG repeating units with two terminal hydroxyl groups. PEGs are classified by their molecular weights; for example, PEG 2000 has an average molecular weight of about 2,000 daltons, and PEG 5000 has an average molecular weight of about 5,000 daltons. PEGs are commercially available from Sigma Chemical Co. and other companies and include, for example, the following: monomethoxypolyethylene glycol (MePEGOH), monomethoxypolyethylene glycolsuccinate (MePEGS), monomethoxypolyethylene glycolsuccinimidyl succinate (MePEG-S-NHS), monomethoxypolyethylene glycolamine (MePEG-NH2), monomethoxypolyethylene glycoltresylate (MePEG-TRES), and monomethoxypolyethylene glycolimidazolylcarbonyl (MePEG-IM). Other PEGs such as those described in U.S. Patent Nos.6,774,180 and 7,053,150 (e.g., mPEG (20 KDa) amine) are also useful for preparing the PEG-lipid conjugates of the present disclosure. The disclosures of these patents are herein incorporated by reference in their entirety for all purposes. In addition, monomethoxypolyethyleneglycolacetic acid (MePEG-CH2COOH) is particularly useful for preparing PEG-lipid conjugates including, e.g., PEG-DAA conjugates. In certain embodiments, the PEG-lipid conjugate or ATTA-lipid conjugate is used together with a CPL. The conjugated lipid that inhibits aggregation of particles may comprise a PEG-lipid including, e.g., a PEG-diacylglycerol (DAG), a PEG dialkyloxypropyl (DAA), a PEG-phospholipid, a PEG-ceramide (Cer), or mixtures thereof. The PEGDAA conjugate may be PEG-dilauryloxypropyl (C12), a PEG-dimyristyloxypropyl (C14), a PEG- dipalmityloxypropyl (C16), a PEG-distearyloxypropyl (C18), or mixtures thereof. 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 - 38 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) for all purposes. Yet additional PEG-lipid conjugates suitable for use in the disclosure include, without limitation, l-[8’-(l,2-dimyristoyl-3-propanoxy)-carboxamido-3’,6’- dioxaoctanyl] carbamoyl-methyl-poly(ethylene glycol) (2 KPEG-DMG). The synthesis of 2 KPEG-DMG is described in U.S. Patent No.7,404,969, the disclosure of which is herein incorporated by reference in its entirety for all purposes. The PEG moiety of the PEG-lipid conjugates described herein may comprise an average molecular weight ranging from about 550 daltons to about 10,000 daltons. In certain instances, the PEG moiety has an average molecular weight of from about 750 daltons to about 5,000 daltons (e.g., from about 1,000 daltons to about 5,000 daltons, from about 1,500 daltons to about 3,000 daltons, from about 750 daltons to about 3,000 daltons, from about 750 daltons to about 2,000 daltons, etc.). In some embodiments, the PEG moiety has an average molecular weight of about 2,000 daltons or about 750 daltons. In addition to the foregoing, it will be readily apparent to those of skill in the art that other hydrophilic polymers can be used in place of PEG. Examples of suitable polymers that can be used in place of PEG include, but are not limited to, polyvinylpyrrolidone, polymethyloxazoline, polyethyloxazoline, polyhydroxypropyl methacrylamide, polymethacrylamide and polydimethylacrylamide, polylactic acid, polyglycolic acid, and derivatized celluloses such as hydroxymethylcellulose or hydroxyethylcellulose. In addition to the foregoing components, the particles (e.g., LNP) of the present disclosure can further comprise cationic poly(ethylene glycol) (PEG) lipids or CPLs (e.g., Chen et al., Bioconj. Chem., 11:433-437 (2000)). Suitable SPLPs and SPLP-CPLs for use in the present disclosure, and methods of making and using SPLPs and SPLP-CPLs, are disclosed, e.g., in U.S. Patent No.6,852,334 and PCT Publication No. WO 00 / 62813, the disclosures of which are herein incorporated by reference in their entirety for all purposes. In certain instances, the conjugated lipid that inhibits aggregation of particles (e.g., PEG-lipid conjugate) may comprise from about 0.1 mol% to about 2 mol%, from about 0.5 mol% to about 2 mol%, from about 1 mol% to about 2 mol%, from about 0.6 mol% to about 1.9 mol%, from about 0.7 mol% to about 1.8 mol%, from about 0.8 mol% to about 1.7 mol%, from about 1 mol% to about 1.8 mol%, from about 1.2 mol% to about 1.8 mol%, from about 1.2 mol% to about 1.7 mol%, from about 1.3 mol% to about 1.6 mol%, from about 1.4 mol% 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 - 39 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 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 In one aspect, the present disclosure provides a lipid nanoparticle (LNP) composition. In certain embodiments, the composition comprises (a) at least one ionizable lipid of formula (I). In certain embodiments, the composition comprises (b) at least one helper lipid. In certain embodiments, the composition comprises (c) at least one cholesterol lipid. In certain embodiments, the composition comprises (d) at least one polymer-conjugated lipid. In certain embodiments, the at least one ionizable 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, 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, 90, 91, 92, 93, 94, 95, 96, 97, 98, or about 99 mol% of the LNP. In certain embodiments, the at least one ionizable lipid comprises less than about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 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, 90, 91, 92, 93, 94, 95, 96, 97, 98, or about 99 mol% of the LNP. In certain embodiments, the at least one ionizable lipid comprises more 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, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, - 40 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 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, 90, 91, 92, 93, 94, 95, 96, 97, 98, or about 99 mol% of the LNP. In certain embodiments, the at least one ionizable lipid comprises about 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, or about 60 mol% of the LNP. In certain embodiments, the at least one ionizable lipid comprises less than about 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, or about 60 mol% of the LNP. In certain embodiments, the at least one ionizable lipid comprises more than about 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, or about 60 mol% of the LNP. In certain embodiments, the at least one ionizable lipid comprises about 30.6 mol% of the LNP. In certain embodiments, the at least one ionizable lipid comprises about 35.0 mol% of the LNP. In certain embodiments, the at least one ionizable lipid comprises about 36.8 mol% of the LNP. In certain embodiments, the at least one ionizable lipid comprises about 40.0 mol% of the LNP. In certain embodiments, the at least one ionizable lipid comprises about 50.0 mol% of the LNP. In certain embodiments, the at least one helper 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, or about 30 mol% of the LNP. In certain embodiments, the at least one helper lipid comprises less than about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or about 30 mol% of the LNP. In certain embodiments, the at least one helper lipid comprises more 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, or about 30 mol% of the LNP. In certain embodiments, the at least one helper lipid comprises about 10 mol% of the LNP. In certain embodiments, the at least one helper lipid comprises about 16.0 mol% of the LNP. In certain embodiments, the at least one helper lipid comprises about 17.5 mol% of the LNP. In certain embodiments, the at least one helper lipid comprises about 21.0 mol% of the LNP. In certain embodiments, the helper lipid is dioleoylphosphatidylethanolamine (DOPE). In certain embodiments, the cholesterol comprises about 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, or about 60 - 41 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) mol% of the LNP. In certain embodiments, the cholesterol comprises less than about 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, or about 60 mol% of the LNP. In certain embodiments, the cholesterol comprises more than about 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, or about 60 mol% of the LNP. In certain embodiments, the cholesterol comprises about 38.5 mol% of the LNP. In certain embodiments, the cholesterol comprises about 40.4 mol% of the LNP. In certain embodiments, the cholesterol comprises about 46.5 mol% of the LNP. In certain embodiments, the cholesterol comprises about 48.5 mol% of the LNP. In certain embodiments, the cholesterol comprises about 50.4 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, or about 10.0 mol% of the LNP. 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, or about 10.0 mol% of the LNP. In certain embodiments, the at least one polymer-conjugated lipid comprises more 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, or about 10.0 mol% of the LNP. In certain embodiments, the at least one polymer-conjugated lipid comprises about 1.5 mol% of the LNP. In certain embodiments, the at least one polymer-conjugated lipid comprises about 1.8 mol% of the LNP. In certain embodiments, the at least one polymer- conjugated lipid comprises about 2.5 mol% of the LNP. In certain embodiments, the one polymer-conjugated lipid and / or a modified derivative thereof comprises 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol- - 42 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 2000 (DMG-PEG-2000): . In about a : :(c):(d) of about 30.6:17.5:50.4:1.5. 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 has a molar ratio of (a):(b):(c):(d) of about 40:10:48.5:1.5. In certain embodiments, the LNP has a molar ratio of (a):(b):(c):(d) of about 50:10:38.5:1.5. In certain embodiments, the LNP further comprises at least one 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 comprises a nucleic acid. In certain embodiments, the nucleic acid cargo 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 cargo is at least partially encapsulated in the LNP. In certain embodiments, the ionizable lipid and nucleic acid cargo have a weight ratio ranging from about 20:1, 19:1, 18:1, 17:1, 16:1, 15:1, 14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, or about 5:1 (ionizable lipid:cargo (w / w)). In certain embodiments, the nucleic acid cargo comprises mRNA. In certain embodiments, the nucleic acid cargo further comprises sgRNA. In certain embodiments, mRNA comprises Cas9 mRNA. In certain embodiments, sRNA comprises TTR sgRNA. Methods Synthetic Methods In another aspect, the present disclosure provides a method of preparing the ionizable lipid compound of formula (I): (I), wherein: - 43 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) A is selected from the group consisting ; R1aand R1bare each independently ; 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); R3aand R3bare each independently selected from the group consisting of optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C6heteroalkyl, optionally substituted C3-C6heteroalkenyl, and optionally substituted C2-C8 heterocycloalkyl; each occurrence of R4aand R4bis independently H; each occurrence of R5ais independently selected from the group consisting of optionally substituted C1-C24alkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C2-C24 heteroalkyl, optionally substituted C2-C12 heterocycloalkyl, optionally substituted C2-C24alkylenyl, optionally substituted C3-C24heteroalkylenyl, optionally substituted C2-C24 alkynylenyl, and optionally substituted C3-C24 heteroalkynylenyl; each occurrence of R5bis H; each occurrence of R6is independently selected from the group consisting of optionally substituted C1-C24alkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C2-C24 heteroalkyl, optionally substituted C2-C12 heterocycloalkyl, optionally substituted C2-C24alkylenyl, optionally substituted C3-C24heteroalkylenyl, optionally substituted C2-C24 alkynylenyl, and optionally substituted C3-C24 heteroalkynylenyl; L is selected from the group consisting of optionally substituted C2-C6alkylenyl and optionally substituted C2-C8 heteroalkylenyl; and Raand Rbare each independently selected from the group consisting of H and optionally substituted C1-C6 alkyl; the method comprising: (a) contacting an amine compound of formula (A1) or (A2): - 44 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) at least two epoxide compounds of formula (B): , to form an alkoxide (b) contacting the an activated acid compound of formula (C): , wherein X is a In certain embodiments, the and epoxide have a molar ratio of about 1:2.4 (ratio of (A1) and (A2):(B)). In certain embodiments, the contacting in step (a) occurs at a temperature of about 80 °C. In certain embodiments, the contacting in step (a) occurs for a period of about 48 h. In certain embodiments, the alkoxide intermediate is dissolved in a solvent before contacting in step (b), optionally wherein the solvent is dichloromethane. In certain embodiments, the contacting in step (b) occurs in the present of a base, optionally wherein the base is a trialkylamine, optionally wherein the trialkylamine is triethylamine. In certain embodiments, X is Cl. In certain embodiments, the amine is selected from the group consisting of N1,N1- dimethylpropane-1,3-diamine, methylamine, ethylamine, 1,1-dimethylhydrazine, 2- methoxyethan-1amine, 2-methoxyethan-1-amine, 2-methoxyethan-1-amine, N1,N1- dimethylethane-1,2-diamine, N1,N1-dimethylbutane-1,4-diamine, N1,N1-diethylethane-1,2- diamine, N1,N1-diethylpropane-1,3-diamine, 2-(pyrrolidin-1-yl)ethan-1-amine, 3-(pyrrolidin- 1-yl)propan-1-amine, 3-(1H-imidazol-1-yl)propan-1-amine, 2-(piperidin-1-yl)ethan-1-amine, 3-(4-methylpiperazin-1-yl)propan-1-amine, N1,N2-dimethylethane-1,2-diamine, N1,N3- dimethylpropane-1,3-diamine, N1,N2-diethylethane-1,2-diamine, N1,N3-dimethyl-N1-(3- (methylamino)propyl)propane-1,3-diamine, and piperazine. In certain embodiments, the epoxide is selected from the group consisting of 2- butyloxirane, 2-pentyloxirane, 2-hexyloxirane, 2-heptyloxirane, 2-octyloxirane, 2- nonyloxirane, 2-decyloxirane, 2-undecyloxirane, and 2-dodecyloxirane. In certain embodiments, the activated acid is selected from the group consisting of hexanoyl chloride, heptanoyl chloride, octanoyl chloride, nonanoyl chloride, decanoyl chloride, undecanoyl chloride, dodecanoyl chloride, tridecanoyl chloride, and tetradecanoyl chloride. - 45 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) Therapeutic Methods In another aspect, the present disclosure provides a method of treating, preventing, and / or ameliorating a disease or disorder in a subject. In certain embodiments, the method comprises administering to the subject at least one lipid nanoparticle (LNP) composition of the present disclosure. In certain embodiments, the LNP comprises (a) at least one ionizable lipid compound of formula (I). In certain embodiments, the LNP comprises (b) at least one helper lipid. In certain embodiments, the LNP comprises (c) at least one cholesterol lipid. In certain embodiments, the LNP comprises (d) at least one polymer-conjugated lipid and / or a modified derivative thereof. In certain embodiments, the LNP comprises (e) at least one cargo molecule. In certain embodiments, LNP is formulated as a pharmaceutical composition comprising at least one pharmaceutically acceptable carrier. In certain embodiments, the disease is cancer. In certain embodiments, the cancer is at least one selected from the group consisting of pancreatic cancer, colorectal cancer, bladder cancer, breast cancer, prostate cancer, renal cancer, hepatocellular cancer, lung cancer, ovarian cancer, cervical cancer, gastric cancer, esophageal cancer, head and neck cancer, melanoma, neuroendocrine cancer, CNS cancer, brain cancer, bone cancer, soft tissue sarcoma, non-small cell lung cancer, small-cell lung cancer, or colon cancer. In certain embodiments, the subject is further administered at least one additional agent or therapy useful for treating, preventing, and / or ameliorating cancer in the subject. In certain embodiments, the subject is a mammal. In certain embodiments, the mammal is a human. Anti-Cancer Agents In some embodiments, the at least one additional agent is an anti-cancer agent. Any suitable anti-cancer agent may be used in the compositions and methods of the present disclosure. The selection of a suitable anti-cancer agent may depend upon, among other things, the type of cancer to be treated and the nanoparticle compositions of the present disclosure. In certain embodiments, the anti-cancer agent may be effective for treating one or more of pancreatic cancer, esophageal cancer, rectal cancer, colon cancer, prostate cancer, kidney cancer, liver cancer, breast cancer, ovarian cancer, and stomach cancer. Examples of anti-cancer agents include, but are not limited to, chemotherapeutic agents, antiproliferative - 46 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) agents, anti-tumor agents, checkpoint inhibitors, and anti-angiogenic agents. For example, In some embodiments, the anti-cancer agent is gemcitabine, doxorubicin, 5-Fu, tyrosine kinase inhibitors, sorafenib, trametinib, rapamycin, fulvestrant, ezalutamide, or paclitaxel. Chemotherapeutic agents include cytotoxic agents (e.g., 5-fluorouracil, cisplatin, carboplatin, methotrexate, daunorubicin, doxorubicin, vincristine, vinblastine, oxorubicin, carmustine (BCNU), lomustine (CCNU), cytarabine USP, cyclophosphamide, estramucine phosphate sodium, altretamine, hydroxyurea, ifosfamide, procarbazine, mitomycin, busulfan, cyclophosphamide, mitoxantrone, carboplatin, cisplatin, interferon alfa-2a recombinant, paclitaxel, teniposide, and streptozoci), cytotoxic alkylating agents (e.g., busulfan, chlorambucil, cyclophosphamide, melphalan, or ethylesulfonic acid), alkylating agents (e.g., asaley, AZQ, BCNU, busulfan, bisulphan, carboxyphthalatoplatinum, CBDCA, CCNU, CHIP, chlorambucil, chlorozotocin, cis-platinum, clomesone, cyanomorpholinodoxorubicin, cyclodisone, cyclophosphamide, dianhydrogalactitol, fluorodopan, hepsulfam, hycanthone, iphosphamide, melphalan, methyl CCNU, mitomycin C, mitozolamide, nitrogen mustard, PCNU, piperazine, piperazinedione, pipobroman, porfiromycin, spirohydantoin mustard, streptozotocin, teroxirone, tetraplatin, thiotepa, triethylenemelamine, uracil nitrogen mustard, and Yoshi-864), antimitotic agents (e.g., allocolchicine, Halichondrin M, colchicine, colchicine derivatives, dolastatin 10, maytansine, rhizoxin, paclitaxel derivatives, paclitaxel, thiocolchicine, trityl cysteine, vinblastine sulfate, and vincristine sulfate), plant alkaloids (e.g., actinomycin D, bleomycin, L-asparaginase, idarubicin, vinblastine sulfate, vincristine sulfate, mitramycin, mitomycin, daunorubicin, VP-16-213, VM-26, navelbine and taxotere), biologicals (e.g., alpha interferon, BCG, G-CSF, GM-CSF, and interleukin-2), topoisomerase I inhibitors (e.g., camptothecin, camptothecin derivatives, and morpholinodoxorubicin), topoisomerase II inhibitors (e.g., mitoxantron, amonafide, m-AMSA, anthrapyrazole derivatives, pyrazoloacridine, bisantrene HCL, daunorubicin, deoxydoxorubicin, menogaril, N,N-dibenzyl daunomycin, oxanthrazole, rubidazone, VM-26 and VP-16), and synthetics (e.g., hydroxyurea, procarbazine, o,p’-DDD, dacarbazine, CCNU, BCNU, cis- diamminedichloroplatimun, mitoxantrone, CBDCA, levamisole, hexamethylmelamine, all- trans retinoic acid, gliadel and porfimer sodium). Antiproliferative agents are compounds that decrease the proliferation of cells. Antiproliferative agents include alkylating agents, antimetabolites, enzymes, biological response modifiers, miscellaneous agents, hormones and antagonists, androgen inhibitors (e.g., flutamide and leuprolide acetate), antiestrogens (e.g., tamoxifen citrate and analogs thereof, toremifene, droloxifene and roloxifene), Additional examples of specific - 47 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) antiproliferative agents include, but are not limited to levamisole, gallium nitrate, granisetron, sargramostim strontium-89 chloride, filgrastim, pilocarpine, dexrazoxane, and ondansetron. The inhibitors of the invention can be administered alone or in combination with other anti-tumor agents, including cytotoxic / antineoplastic agents and anti-angiogenic agents. Cytotoxic / anti-neoplastic agents are defined as agents which attack and kill cancer cells. Some cytotoxic / anti-neoplastic agents are alkylating agents, which alkylate the genetic material in tumor cells, e.g., cis-platin, cyclophosphamide, nitrogen mustard, trimethylene thiophosphoramide, carmustine, busulfan, chlorambucil, belustine, uracil mustard, chlomaphazin, and dacabazine. Other cytotoxic / anti-neoplastic agents are antimetabolites for tumor cells, e.g., cytosine arabinoside, fluorouracil, methotrexate, mercaptopuirine, azathioprime, and procarbazine. Other cytotoxic / anti-neoplastic agents are antibiotics, e.g., doxorubicin, bleomycin, dactinomycin, daunorubicin, mithramycin, mitomycin, mytomycin C, and daunomycin. There are numerous liposomal formulations commercially available for these compounds. Still other cytotoxic / anti-neoplastic agents are mitotic inhibitors (vinca alkaloids). These include vincristine, vinblastine and etoposide. Miscellaneous cytotoxic / anti- neoplastic agents include taxol and its derivatives, L-asparaginase, anti-tumor antibodies, dacarbazine, azacytidine, amsacrine, melphalan, VM-26, ifosfamide, mitoxantrone, and vindesine. Anti-angiogenic agents are well known to those of skill in the art. Suitable anti- angiogenic agents for use in the methods and compositions of the present disclosure include anti-VEGF antibodies, including humanized and chimeric antibodies, anti-VEGF aptamers and antisense oligonucleotides. Other known inhibitors of angiogenesis include angiostatin, endostatin, interferons, interleukin 1 (including alpha and beta) interleukin 12, retinoic acid, and tissue inhibitors of metalloproteinase-1 and -2. (TIMP-1 and -2). Small molecules, including topoisomerases such as razoxane, a topoisomerase II inhibitor with anti-angiogenic activity, can also be used. Other anti-cancer agents that can be used in combination with the disclosed compounds include, but are not limited to: acivicin; aclarubicin; acodazole hydrochloride; acronine; adozelesin; aldesleukin; altretamine; ambomycin; ametantrone acetate; aminoglutethimide; amsacrine; anastrozole; anthramycin; asparaginase; asperlin; azacitidine; azetepa; azotomycin; batimastat; benzodepa; bicalutamide; bisantrene hydrochloride; bisnafide dimesylate; bizelesin; bleomycin sulfate; brequinar sodium; bropirimine; busulfan; cactinomycin; calusterone; caracemide; carbetimer; carboplatin; carmustine; carubicin hydrochloride; carzelesin; cedefingol; chlorambucil; cirolemycin; cisplatin; cladribine; - 48 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) crisnatol mesylate; cyclophosphamide; cytarabine; dacarbazine; dactinomycin; daunorubicin hydrochloride; decitabine; dexormaplatin; dezaguanine; dezaguanine mesylate; diaziquone; docetaxel; doxorubicin; doxorubicin hydrochloride; droloxifene; droloxifene citrate; dromostanolone propionate; duazomycin; edatrexate; eflornithine hydrochloride; elsamitrucin; enloplatin; enpromate; epipropidine; epirubicin hydrochloride; erbulozole; esorubicin hydrochloride; estramustine; estramustine phosphate sodium; etanidazole; etoposide; etoposide phosphate; etoprine; fadrozole hydrochloride; fazarabine; fenretinide; floxuridine; fludarabine phosphate; fluorouracil; fluorocitabine; fosquidone; fostriecin sodium; gemcitabine; gemcitabine hydrochloride; hydroxyurea; idarubicin hydrochloride; ifosfamide; ilmofosine; interleukin II (including recombinant interleukin II, or rIL2), interferon alfa-2a; interferon alfa-2b; interferon alfa-n1; interferon alfa-n3; interferon beta-I a; interferon gamma-I b; iproplatin; irinotecan hydrochloride; lanreotide acetate; letrozole; leuprolide acetate; liarozole hydrochloride; lometrexol sodium; lomustine; losoxantrone hydrochloride; masoprocol; maytansine; mechlorethamine hydrochloride; megestrol acetate; melengestrol acetate; melphalan; menogaril; mercaptopurine; methotrexate; methotrexate sodium; metoprine; meturedepa; mitindomide; mitocarcin; mitocromin; mitogillin; mitomalcin; mitomycin; mitosper; mitotane; mitoxantrone hydrochloride; mycophenolic acid; nocodazole; nogalamycin; ormaplatin; oxisuran; paclitaxel; pegaspargase; peliomycin; pentamustine; peplomycin sulfate; perfosfamide; pipobroman; piposulfan; piroxantrone hydrochloride; plicamycin; plomestane; porfimer sodium; porfiromycin; prednimustine; procarbazine hydrochloride; puromycin; puromycin hydrochloride; pyrazofurin; riboprine; rogletimide; safingol; safingol hydrochloride; semustine; simtrazene; sparfosate sodium; sparsomycin; spirogermanium hydrochloride; spiromustine; spiroplatin; streptonigrin; streptozocin; sulofenur; talisomycin; tecogalan sodium; tegafur; teloxantrone hydrochloride; temoporfin; teniposide; teroxirone; testolactone; thiamiprine; thioguanine; thiotepa; tiazofurin; tirapazamine; toremifene citrate; trestolone acetate; triciribine phosphate; trimetrexate; trimetrexate glucuronate; triptorelin; tubulozole hydrochloride; uracil mustard; uredepa; vapreotide; verteporfin; vinblastine sulfate; vincristine sulfate; vindesine; vindesine sulfate; vinepidine sulfate; vinglycinate sulfate; vinleurosine sulfate; vinorelbine tartrate; vinrosidine sulfate; vinzolidine sulfate; vorozole; zeniplatin; zinostatin; zorubicin hydrochloride. Other anti-cancer drugs include, but are not limited to: 20-epi-1,25 dihydroxyvitamin D3; 5-ethynyluracil; abiraterone; aclarubicin; acylfulvene; adecypenol; adozelesin; aldesleukin; ALL-TK antagonists; altretamine; ambamustine; amidox; amifostine; aminolevulinic acid; amrubicin; amsacrine; anagrelide; anastrozole; - 49 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) andrographolide; angiogenesis inhibitors; antagonist D; antagonist G; antarelix; anti- dorsalizing morphogenetic protein-1; antiandrogen, prostatic carcinoma; antiestrogen; antineoplaston; antisense oligonucleotides; aphidicolin glycinate; apoptosis gene modulators; apoptosis regulators; apurinic acid; ara-CDP-DL-PTBA; arginine deaminase; asulacrine; atamestane; atrimustine; axinastatin 1; axinastatin 2; axinastatin 3; azasetron; azatoxin; azatyrosine; baccatin III derivatives; balanol; batimastat; BCR / ABL antagonists; benzochlorins; benzoylstaurosporine; beta lactam derivatives; beta-alethine; betaclamycin B; betulinic acid; bFGF inhibitor; bicalutamide; bisantrene; bisaziridinylspermine; bisnafide; bistratene A; bizelesin; breflate; bropirimine; budotitane; buthionine sulfoximine; calcipotriol; calphostin C; camptothecin derivatives; canarypox IL-2; capecitabine; carboxamide-amino-triazole; carboxyamidotriazole; CaRest M3; CARN 700; cartilage derived inhibitor; carzelesin; casein kinase inhibitors (ICOS); castanospermine; cecropin B; cetrorelix; chlorins; chloroquinoxaline sulfonamide; cicaprost; cis-porphyrin; cladribine; clomifene analogues; clotrimazole; collismycin A; collismycin B; combretastatin A4; combretastatin analogue; conagenin; crambescidin 816; crisnatol; cryptophycin 8; cryptophycin A derivatives; curacin A; cyclopentanthraquinones; cycloplatam; cypemycin; cytarabine ocfosfate; cytolytic factor; cytostatin; dacliximab; decitabine; dehydrodidemnin B; deslorelin; dexamethasone; dexifosfamide; dexrazoxane; dexverapamil; diaziquone; didemnin B; didox; diethylnorspermine; dihydro-5-azacytidine; dihydrotaxol, 9-; dioxamycin; diphenyl spiromustine; docetaxel; docosanol; dolasetron; doxifluridine; droloxifene; dronabinol; duocarmycin SA; ebselen; ecomustine; edelfosine; edrecolomab; eflornithine; elemene; emitefur; epirubicin; epristeride; estramustine analogue; estrogen agonists; estrogen antagonists; etanidazole; etoposide phosphate; exemestane; fadrozole; fazarabine; fenretinide; filgrastim; finasteride; flavopiridol; flezelastine; fluasterone; fludarabine; fluorodaunorunicin hydrochloride; forfenimex; formestane; fostriecin; fotemustine; gadolinium texaphyrin; gallium nitrate; galocitabine; ganirelix; gelatinase inhibitors; gemcitabine; glutathione inhibitors; hepsulfam; heregulin; hexamethylene bisacetamide; hypericin; ibandronic acid; idarubicin; idoxifene; idramantone; ilmofosine; ilomastat; imidazoacridones; imiquimod; immunostimulant peptides; insulin-like growth factor-1 receptor inhibitor; interferon agonists; interferons; interleukins; iobenguane; iododoxorubicin; ipomeanol, 4-; iroplact; irsogladine; isobengazole; isohomohalicondrin B; itasetron; jasplakinolide; kahalalide F; lamellarin-N triacetate; lanreotide; leinamycin; lenograstim; lentinan sulfate; leptolstatin; letrozole; leukemia inhibiting factor; leukocyte alpha interferon; leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole; linear - 50 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) polyamine analogue; lipophilic disaccharide peptide; lipophilic platinum compounds; lissoclinamide 7; lobaplatin; lombricine; lometrexol; lonidamine; losoxantrone; lovastatin; loxoribine; lurtotecan; lutetium texaphyrin; lysofylline; lytic peptides; maitansine; mannostatin A; marimastat; masoprocol; maspin; matrilysin inhibitors; matrix metalloproteinase inhibitors; menogaril; merbarone; meterelin; methioninase; metoclopramide; MIF inhibitor; mifepristone; miltefosine; mirimostim; mismatched double stranded RNA; mitoguazone; mitolactol; mitomycin analogues; mitonafide; mitotoxin fibroblast growth factor-saporin; mitoxantrone; mofarotene; molgramostim; monoclonal antibody, human chorionic gonadotrophin; monophosphoryl lipid A+myobacterium cell wall sk; mopidamol; multiple drug resistance gene inhibitor; multiple tumor suppressor 1-based therapy; mustard anticancer agent; mycaperoxide B; mycobacterial cell wall extract; myriaporone; N-acetyldinaline; N-substituted benzamides; nafarelin; nagrestip; naloxone+pentazocine; napavin; naphterpin; nartograstim; nedaplatin; nemorubicin; neridronic acid; neutral endopeptidase; nilutamide; nisamycin; nitric oxide modulators; nitroxide antioxidant; nitrullyn; O6-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone; ondansetron; ondansetron; oracin; oral cytokine inducer; ormaplatin; osaterone; oxaliplatin; oxaunomycin; paclitaxel; paclitaxel analogues; paclitaxel derivatives; palauamine; palmitoylrhizoxin; pamidronic acid; panaxytriol; panomifene; parabactin; pazelliptine; pegaspargase; peldesine; pentosan polysulfate sodium; pentostatin; pentrozole; perflubron; perfosfamide; perillyl alcohol; phenazinomycin; phenylacetate; phosphatase inhibitors; picibanil; pilocarpine hydrochloride; pirarubicin; piritrexim; placetin A; placetin B; plasminogen activator inhibitor; platinum complex; platinum compounds; platinum- triamine complex; porfimer sodium; porfiromycin; prednisone; propyl bis-acridone; prostaglandin J2; proteasome inhibitors; protein A-based immune modulator; protein kinase C inhibitor; protein kinase C inhibitors, microalgal; protein tyrosine phosphatase inhibitors; purine nucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine; pyridoxylated hemoglobin polyoxyethylene conjugate; raf antagonists; raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors; ras inhibitors; ras-GAP inhibitor; retelliptine demethylated; rhenium Re 186 etidronate; rhizoxin; ribozymes; RII retinamide; rogletimide; rohitukine; romurtide; roquinimex; rubiginone B1; ruboxyl; safingol; saintopin; SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics; semustine; senescence derived inhibitor 1; sense oligonucleotides; signal transduction inhibitors; signal transduction modulators; single chain antigen binding protein; sizofuran; sobuzoxane; sodium borocaptate; sodium phenylacetate; solverol; somatomedin binding protein; sonermin; sparfosic acid; spicamycin D; - 51 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) spiromustine; splenopentin; spongistatin 1; squalamine; stem cell inhibitor; stem-cell division inhibitors; stipiamide; stromelysin inhibitors; sulfinosine; superactive vasoactive intestinal peptide antagonist; suradista; suramin; swainsonine; synthetic glycosaminoglycans; tallimustine; tamoxifen methiodide; tauromustine; tazarotene; tecogalan sodium; tegafur; tellurapyrylium; telomerase inhibitors; temoporfin; temozolomide; teniposide; tetrachlorodecaoxide; tetrazomine; thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetic; thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroid stimulating hormone; tin ethyl etiopurpurin; tirapazamine; titanocene bichloride; topsentin; toremifene; totipotent stem cell factor; translation inhibitors; tretinoin; triacetyluridine; triciribine; trimetrexate; triptorelin; tropisetron; turosteride; tyrosine kinase inhibitors; tyrphostins; UBC inhibitors; ubenimex; urogenital sinus-derived growth inhibitory factor; urokinase receptor antagonists; vapreotide; variolin B; vector system, erythrocyte gene therapy; velaresol; veramine; verdins; verteporfin; vinorelbine; vinxaltine; vitaxin; vorozole; zanoterone; zeniplatin; zilascorb; and zinostatin stimalamer. In some embodiments, the anti- cancer drug is 5-fluorouracil, taxol, or leucovorin. In some embodiments, the anti-cancer agent may be a prodrug form of an anti-cancer agent. As used herein, the term “prodrug form” and its derivatives is used to refer to a drug that has been chemically modified to add and / or remove one or more substituents in such a manner that, upon introduction of the prodrug form into a subject, such a modification may be reversed by naturally occurring processes, thus reproducing the drug. The use of a prodrug form of an anti-cancer agent in the compositions, among other things, may increase the concentration of the anti-cancer agent in the compositions of the present disclosure. In certain embodiments, an anti-cancer agent may be chemically modified with an alkyl or acyl group or some form of lipid. The selection of such a chemical modification, including the substituent(s) to add and / or remove to create the prodrug, may depend upon a number of factors including, but not limited to, the particular drug and the desired properties of the prodrug. One of ordinary skill in the art, with the benefit of this disclosure, will recognize suitable chemical modifications. 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. - 52 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 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 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. - 53 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 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 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 - 54 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 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, 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- - 55 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 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, 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” - 56 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 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 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 - 57 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 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 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 - 58 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 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 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 - 59 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 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 (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 - 60 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 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 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 - 61 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 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 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 Materials Amines, epoxides, acyl chlorides, TEA were purchased from Sigma Aldrich, Tokyo Chemical Industry (TCI) and Ambeed. Alanine transaminase (ALT) colorimetric activity assay kit and aspartate aminotransferase (AST) colorimetric activity assay kit were purchased from Cayman Chemical.1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2- distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dimyristoyl-rac-glycero-3- methoxypolyethylene glycol-2000 (DMG-PEG 2000) and cholesterol were obtained from Avanti Polar Lipids. DLin-MC3-DMA was purchased from MedChem Express. Cas9 mRNA - 62 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) (5moU) was bought from TriLink. Highly modified sgRNA target mouse TTR (guide No. G211) was chemically synthesized by a commercial supplier (AxoLabs) according to methods known to those of ordinary skill in the art. Anhydrous DCM, Porcine liver esterase, amiloride hydrochloride, chlorpromazine, genistein, methyl-beta-cyclodextrin and TTR siRNA pool (SASI_Mm01_00076059, SASI_Mm01_00076060 and SASI_Mm01_00076061) were purchased from Sigma Aldrich. mRNA Synthesis Codon optimized firefly Luciferase sequence and human FGF21 sequence was cloned into an mRNA production plasmid (optimized 3’ and 5’ UTR with a 101 polyA tail), in vitro transcribed in the presence of 1-methyl pseudouridine modified nucleoside, co- transcriptionally capped using the CleanCapTM technology (TriLink) and cellulose purified to remove double-stranded RNAs. Purified mRNA was ethanol precipitated, washed, re- suspended in nuclease-free water, and subjected to quality control. All mRNAs were stored at -20 °C until use. General methods for the synthesis of SRB-lipidpoids SRB-lipidoids were synthesized using a one-pot, two-step method. First, amine (0.1 mmol, 1 equiv.) and epoxide (0.24 mmol, 2.4 equiv.) were combined in a glass vial and heated at 80 °C for 48 h. Next, the reactant was dissolved in 2 mL anhydrous DCM at RT, followed by the addition of acyl chloride (0.24 mmol, 2.4 equiv.) and TEA (0.3 mmol, 3 equiv.).12 h later, DCM and TEA were removed under vacuum and crude SRB-lipidoids were dissolved in EtOH for initial screening. Notably, if an amine was in the salt form, excessive TEA was added to neutralize it in the first step. All crude SRB-lipidoids were confirmed by mass spectrometry. Purification and characterization of 11-10-8 To purify the top-performing SRB-lipidoid 11-10-8, its crude product was separated using a CombiFlash NextGen 300+ chromatography system with gradient elution from CH2Cl2 to 75:22:3 CH2Cl2 / MeOH / NH4OH (aq), and the desired fractions were collected as light brown oil (82% yield).11-10-8 was characterized by mass spectrometry (MS) and nuclear magnetic resonance spectroscopy (NMR). MS-ESI: calculated for C42H82N2O4: 678.63, found [M+H]+ = 679.62;1H NMR (400 MHz, MeOD) δ: 4.95 (tt, J = 8.2, 4.3 Hz, 2H), 3.19 – 3.08 (m, 2H), 2.84 (t, J = 6.9 Hz, 2H), 2.72 (dd, J = 14.0, 7.7 Hz, 2H), 2.62 (ddd, - 63 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) J = 14.0, 10.3, 4.0 Hz, 2H), 2.45 – 2.28 (m, 4H), 2.14 – 2.04 (m, 4H), 1.72 – 1.48 (m, 8H), 1.42 – 1.26 (m, 44H), 0.93 (h, J = 3.1 Hz, 12H). LNP formulation and optimization For initial in vitro and in vivo screening, SRB-LNPs were prepared by pipette mixing of the ethanolic phase containing SRB-lipidoid, DOPE, cholesterol and DMG-PEG with an aqueous phase (10 mM citrate buffer, pH 3) containing mRNA at a volume ratio of 1:3 and then diluted in culture medium or 1× PBS for cell or animal treatment, respectively. The weight ratio of SRB-lipidoid, DOPE, cholesterol, DMG-PEG and mRNA was fixed at 16:10:10:3:1.6. To formulate 11-10-8 SRB-LNP by microfluidic mixing, the ethanolic phase containing lipids was mixed with the aqueous phase containing mRNA at a flow rate ratio of 1:3 and at a 11-10-8 / mRNA weight ratio of 10:1 in a microfluidic chip device. To optimize the formulation of 11-10-8 SRB-LNP, various LNPs formulated by microfluidic mixing were tested in vivo, and the optimal one (11-10-8 / DOPE / Chol / DMG-PEG = 40:10:48.8:1.5) was chosen for subsequent studies. The benchmark MC3 LNP was formulated with MC3, DSPC, cholesterol and DMG-PEG at a molar ratio of 50:10:38.5:1.5 using microfluidic mixing at a MC3 / mRNA weight ratio of 10:1. LNPs were dialyzed against 1× PBS in a 20 kDa MWCO cassette for 2 h, filtered through a 0.22 μM filter and stored at 4 °C. Characterization 1H NMR were recorded using a Bruker 400 MHz NMR spectrometer. LC-MS was performed on a Waters Acquity LCMS system equipped with UV-Vis and MS detectors. The hydrodynamic size, polydispersity index (PDI) and zeta potential of LNPs were measured using a Malvern Zetasizer Nano ZS90. The morphology of LNPs was characterized by a cryo-electron microscope (Titan Krios, Thermo Fisher). The mRNA encapsulation efficiency and the pKa of LNP were determined using a modified Quant-iT RiboGreen RNA assay (Invitrogen) and a 6-(p-toluidinyl)naphthalene-2-sulfonic acid (TNS) assay, respectively. Cell culture and animal studies Human hepatocellular carcinoma HepG2 cell line was purchased from American Type Culture Collection (ATCC) and maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 U / mL penicillin and 100 μg / mL streptomycin. Cells were cultured at 37 °C in a humidified incubator of 5% CO2, and - 64 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) routinely tested for mycoplasma contamination. C57BL / 6 mice (6-8 weeks, 18-20 g) were purchased from Jackson Laboratory. High-throughput in vitro and in vivo screening of SRB-LNPs HepG2 cells were seeded in 96-well plates at a density of 5,000 per well overnight and mLuc-loaded SRB-LNPs (15 ng / well) were used to treat cells for 24 h. Luciferase expression was evaluated by Luciferase Reporter 1000 Assay System (Promega, #E4550) and cell viability was measured using a CellTiter-Glo Luminescent Cell Viability Assay (Promega, #G7572) according to manufacturer’s protocols. For in vivo screening, mice were i.v. injected with mLuc-loaded SRB-LNPs at an mRNA dose of 0.1 mg / kg.4 h later, mice were intraperitoneally (i.p.) injected with D-luciferin potassium salt (150 mg / kg), and bioluminescence imaging was performed using an in vivo imaging system (PerkinElmer). Molecular dynamics simulations Lipid dynamics simulations of 11-10-8 and MC3 were optimized at CHARMm force field. The lipid simulations were run up to 4000 steps to achieve the energy-minimized structures. All bonds containing intermolecular interactions were constrained using the Smart Minimizer algorithm (i.e., 1000 steps of Steepest Descent with Root-Mean-Squared (RMS) gradient of 3 and Conjugate Gradient minimization). The overall RMS gradient tolerance was set to 0.01. Momany-Rone method was assigned for partial charge estimation using Discovery Studio 2018 (Accelrys)44. The dimensionless packing parameter P of a lipid molecule was calculated as P = V / (AL) based on its Van der Waals molecule volume (V), cross section area of polar head (A) and average tail length (L). V, A and L were derived from the optimized conformation of the lipid as well as the estimated atomic Van der Waals radius. Hemolysis assay Mouse red blood cells (RBCs) were isolated and washed three times with 1× PBS by centrifugation at 700 g for 5 min. Next, RBCs were diluted to a 4% vol / vol RBC solution either in neutral (pH 7.4) or acidic PBS (pH 6.0), and incubated with LNPs at a final mRNA concentration of 3 μg / mL at 37 °C for 1 h. Finally, the RBC solution was centrifuged at 700 g for 5 min and 100 μL supernatant was transferred into a 96-well plate. The absorption at 540 nm was determined with a plate reader. Positive and negative controls were carried out with 0.1% Triton-X and 1× PBS, respectively. - 65 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) Cellular internalization inhibition HepG2 cells were seeded in a 96-well plate at a density of 5,000 per well overnight. Cells were pre-treated with 5 mM amiloride, 20 μM chlorpromazine, 0.2 mM genistein or 5 mM Mβ-CD for 30 min. Then, cells were treated with mLuc-loaded LNPs (15 ng / well) for 24 h. Luciferase expression was determined as described elsewhere herein. Systemic mRNA delivery For GFP mRNA delivery, mice were i.v. injected with GFP mRNA-loaded LNPs at an mRNA dose of 0.25 mg / kg.4 h later, mice were euthanized and major organs (heart, liver, spleen, lung and kidneys) were collected for ex vivo fluorescence imaging. For FGF21 mRNA delivery, mice were i.v. injected with FGF21 mRNA-loaded LNPs at an mRNA dose of 0.25 mg / kg. Serum was collected at 0, 6, 30 and 48 h post-injection and analyzed by ELISA (R&D, #DF2100). For Cas9 mRNA / TTR sgRNA co-delivery, mice were i.v. injected with Cas9 mRNA / sgRNA (4:1, wt:wt)-loaded LNPs at a total RNA dose of 1 mg / kg. Serum was collected at on day 0, 7, 28 and analyzed by ELISA (Aviva Systems Biology, #OKIA00111). Some of mice were euthanized on day 7, and livers were collected to determine the on-target indel frequency by next-generation sequencing (NGS) as describe previously. Systemic siRNA delivery Mice were i.v. injected with TTR siRNA-loaded LNPs at an siRNA dose of 1 mg / kg. Serum was collected at 0, 3, 7, 14, 21 and 28 days post-injection and analyzed by ELISA. To determine the ED50, mice were i.v. injected with TTR siRNA-loaded LNPs at siRNA doses of 0.01, 0.1 and 1 mg / kg. On day 3, serum was collected for ELISA analysis. Safety evaluation Serum was collected at 24 h post-treatment of Cas9 mRNA / TTR sgRNA-loaded LNPs (1 mg / kg). Liver function was evaluated by measuring serum AST and ALT activities. Statistical analysis Data are presented as mean ± SD. Student’s t-test or one-way analysis of variance (ANOVA) followed by Tukey test was applied for comparison between two groups or among multiple groups, respectively. p < 0.05 was considered to be statistically significant. - 66 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) FGF21 mRNA therapy in obese mice Male mice were fed a HFD (#MP290194410, MP Biomedicals, Santa Ana, California, USA) for three weeks to induce obesity (body weight ~30g) and fatty liver41. These obese mice were i.v. injected with FGF21 mRNA-loaded LNPs at an mRNA dose of 0.25 mg / kg. Serum was collected at 0, 3, 6, 12, 24 and 48 h post-injection and analyzed by ELISA (R&D, #DF2100). For the therapeutic study, obese mice were i.v. injected with various LNP formulations at an mRNA dose of 0.25 mg / kg every other day for three doses. Body weights were recorded every two days since the start of the first treatment. Male mice fed with a normal chow diet (NCD) was used a control group. At the end of experiment, mice were anesthetized, and livers were excised, rinsed and weighed. Livers were fixed in 10% neutral formalin overnight, embedded in paraffin, cut into 5 μm sections, and stained with hematoxylin and eosin (H&E) for histological examination of steatosis. The semi-quantitative analysis for liver steatosis was performed according to an adapted scoring protocol. Intramuscular mRNA delivery Mice were i.m. injected with mLuc-loaded LNPs at an mRNA dose of 0.1 mg / kg.4 h later, mice were i.p. injected with D-luciferin potassium salt (150 mg / kg), and bioluminescence imaging was performed using an in vivo imaging system. Example 1: Space shuttle-inspired design and construction of SRB-lipidoids A structure- and function-mimetic strategy was adopted, and the design of SRB- lipidoids was conceptualized based on NASA’s space shuttle (FIGs.1A-1B and FIG.2B). A tandem, hierarchical, and in situ construction method based on a one-pot, two-step, three- component reaction (3-CR) was developed, which enabled rapid and parallel synthesis of degradable SRB-lipidoids. In this approach, conjugation of the first alkyl chain occurs by nucleophilic addition of an amine to an alkyl epoxide to provide a secondary alcohol. In certain embodiments, the amine is a primary amine and the amine undergoes two consecutive epoxide addition reactions to generate a tertiary amine. In other embodiments, the amine is a diamine, wherein the diamine comprises at least two secondary amines. In such embodiments, each secondary amine undergoes a single addition to an epoxide to generate a bis-alkylated diamine comprising two tertiary amines. Regardless of the nature of the amine (i.e., single primary amine vs. secondary - 67 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) diamine), the resulting compound comprises a bis-secondary alcohol which may undergo further derivatization. In certain embodiments, each secondary alcohol is acylated with an acyl halide (e.g., acyl chloride) to provide the lipidoid compounds comprising ester-linked alkyl tails. Since the headgroup (i.e. amine), body tail (i.e., epoxide) and branch tail (i.e., acyl halide) can be independently controlled by using a variety of inexpensive and commercially available materials, this 3-CR is ideal for high-throughput and cost-efficient establishment of a large and systematic combinatorial library of SRB-lipidoids for screening and SAR analysis. Moreover, since (i) both reactions are highly efficient with >80% overall yield, (ii) the solvent dichloromethane (DCM) and base triethylamine (TEA) are easily removed, and (iii) the leftover reactive acyl chloride is quenched in ethanol, the crude SRB-lipidoids can be directly used for LNP formulation and screening. Exemplary amines, epoxides, and acyl chlorides are provided herein (Tables 1-3). Table 1. Exemplary amines Amine Structure Name No. 1 N1,N1-dimethylpropane-1,3-diamine  2 MeNH2 methylamine 3 EtNH2ethylamine 4 N(Me)2NH2 1,1-dimethylhydrazine 4-methylpiperazin-1-amine 5 6 2-methoxyethan-1-amine N1,N1-dimethylethane-1,2-diamine 7 N1,N1-dimethylbutane-1,4-diamine 8 N1,N1-diethylethane-1,2-diamine 9 N1,N1-diethylpropane-1,3-diamine 10 2-(pyrrolidin-1-yl)ethan-1-amine 11 3-(pyrrolidin-1-yl)propan-1-amine 12 - 68 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 3-(1H-imidazol-1-yl)propan-1-amine 13 2-(piperidin-1-yl)ethan-1-amine 14 3-(4-methylpiperazin-1-yl)propan-1-amine 15 N1,N2-dimethylethane-1,2-diamine 16 17N1,N3-dimethylpropane-1,3-diamine N1,N2-diethylethane-1,2-diamine 18 N1,N3-di119methyl-N -(3- (methylamino)propyl)propane-1,3-diamine HN piperazine 20 NH Table 2. Exemplary epoxides Epoxide No. Structure Name 62-butyloxirane 82-hexyloxirane102-octyloxirane 122-decyloxirane 142-dodecyloxirane Table 3. Exemplary acyl chloridesaAcyl Chloride No. Structure Name hexanoyl chloride 6 octanoyl chloride 8 decanoyl chloride 10 dodecanoyl chloride 12 tetradecanoyl chloride 14aany suitable acylating reagent may be used, including but not limited to other acyl halides (e.g., acyl bromides), carboxylic acids, esters, and activated esters (e.g., pentafluorophenoxy esters and hydroxybenzotriazole esters), inter alia. - 69 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) Example 2: Optimizing the tail region of SRB-lipidoids The structure of SRB-lipidoids was systematically optimized via structure-activity- relationship (SAR) investigation. Since the lipidoid comprises three distinct moieties (i.e., headgroup, body tail, and branch tail) that can be altered in SRB-lipidoids (FIG.2A), optimization was initiated with the tail regions, then the head groups, in order to screen a large combinatorial library of SRB-lipidoids in a resource-effective manner. Therefore, in the first library (Library 1), the headgroup was kept constant as amine 1 (i.e., 3-(dimethylamino)- 1-propylamine) and the tail regions (i.e., both body tail and branch tail) were varied (FIG.2B and Table 4). Table 4. Exemplary lipids (Library 1) Nam Compound e 1-6-6 ((3-(dimethylamino)propyl)azanediyl)bis(hexane-1,2-diyl) dihexanoate 1-6-8   ((3-(dimethylamino)propyl)azanediyl)bis(hexane-1,2-diyl) dioctanoate 1-6- 10 ((3-(dimethylamino)propyl)azanediyl)bis(hexane-1,2-diyl) bis(decanoate) 1-6- 12 ((3-(dimethylamino)propyl)azanediyl)bis(hexane-1,2-diyl) didodecanoate 1-6- 14 ((3-(dimethylamino)propyl)azanediyl)bis(hexane-1,2-diyl) ditetradecanoate - 70 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 1-8-6   ((3-(dimethylamino)propyl)azanediyl)bis(octane-1,2-diyl) dihexanoate 1-8-8 ((3-(dimethylamino)propyl)azanediyl)bis(octane-1,2-diyl) dioctanoate 1-8- 10 ((3-(dimethylamino)propyl)azanediyl)bis(octane-1,2-diyl) bis(decanoate) N O O 1-8- 12 O O N ((3-(dimethylamino)propyl)azanediyl)bis(octane-1,2-diyl) didodecanoate 1-8- 14 ((3-(dimethylamino)propyl)azanediyl)bis(octane-1,2-diyl) ditetradecanoate 1-10- 6 ((3-(dimethylamino)propyl)azanediyl)bis(decane-1,2-diyl) dihexanoate 1-10- 8 ((3-(dimethylamino)propyl)azanediyl)bis(decane-1,2-diyl) dioctanoate 1-10- 10 ((3-(dimethylamino)propyl)azanediyl)bis(decane-1,2-diyl) bis(decanoate) - 71 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 1-10- 12 ((3-(dimethylamino)propyl)azanediyl)bis(decane-1,2-diyl) didodecanoate N O O 1-10- 14 O O N ((3-(dimethylamino)propyl)azanediyl)bis(decane-1,2-diyl) ditetradecanoate 1-12- 6 ((3-(dimethylamino)propyl)azanediyl)bis(dodecane-1,2-diyl) dihexanoate 1-12- 8 ((3-(dimethylamino)propyl)azanediyl)bis(dodecane-1,2-diyl) dioctanoate 1-12- 10   ((3-(dimethylamino)propyl)azanediyl)bis(dodecane-1,2-diyl) bis(decanoate) 1-12- 12 ((3-(dimethylamino)propyl)azanediyl)bis(dodecane-1,2-diyl) didodecanoate 1-12- 14 ((3-(dimethylamino)propyl)azanediyl)bis(dodecane-1,2-diyl) ditetradecanoate 1-14- 6 ((3-(dimethylamino)propyl)azanediyl)bis(tetradecane-1,2-diyl) dihexanoate - 72 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 1-14- 8 ((3-(dimethylamino)propyl)azanediyl)bis(tetradecane-1,2-diyl) dioctanoate 1-14- 10 ((3-(dimethylamino)propyl)azanediyl)bis(tetradecane-1,2-diyl) bis(decanoate) 1-14- 12 ((3-(dimethylamino)propyl)azanediyl)bis(tetradecane-1,2-diyl) didodecanoate 1-14- 14 ((3-(dimethylamino)propyl)azanediyl)bis(tetradecane-1,2-diyl) ditetradecanoate Epoxides with short alkyl chains (five variations between 6 and 14 carbons) were used as body tails to minimize the molecular weights of non-degradable parts (< 500 Da) for accelerated physiological clearance (FIG.2A and Table 4). Correspondingly, acyl chlorides with short alkyl chains (five variations between 6 and 14 carbons) were selected as branch tails. The SRB-lipidoids described herein are referred to according to the formula (X-Y-Z) (FIG.2B), wherein X refers to the amine group (Table 1), Y refers to the epoxide-derived fragment (Table 2), and Z refers to the acyl fragment (Table 3). For example, lipidoid “1-6-8” refers to a lipidoid prepared from amine 1, epoxide 6, and acyl chloride 8. The resulting combinatorially synthesized SRB-lipidoids in Library 1 were formulated into 25 SRB-LNPs along with 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), cholesterol (Chol), and 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol- 2000 (DMG-PEG) by pipette mixing (FIG.2B). To evaluate the mRNA delivery potency of SRB-LNPs, 1-methylpseudouridine (m1ψ)-modified firefly luciferase mRNA (mLuc) was encapsulated into these LNPs during formulation, and Luc expression in vitro (HepG2 cells) and in vivo after intravenous (i.v.) injection into C57BL / 6 mice was determined (FIGs.2C- 2D and FIG.3). For all SRB-LNPs, Luc expression was mainly observed in the upper abdomen by in vivo bioluminescence imaging (BLI), which was identified to come from the - 73 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) liver by ex vivo BLI (FIG.4). It was found that the attachment of two branch tails to aminoalcohol lipidoids boosted delivery efficiencies both in vitro and in vivo (FIGs.2C-2D). Generally, the in vitro potencies of SRB-lipidoids were affected by the length of the body tail (FIG.2C), while their in vivo potencies were affected by the length of both tails (FIG.2D). However, there was no correlation between in vitro and in vivo potencies of SRB-lipidoids (FIG.2E), a phenomenon also reported by others. Next, the relationship between tail structure and in vivo potency was investigated. While all aminoalcohol lipidoids tested lacked in vivo efficacy (total flux < 107 p / s), appending branch tails dramatically boosted mRNA delivery efficiency by 1-3 orders of magnitude (FIG.2F). In certain, non-limiting embodiments, 8- or 10-carbon epoxide and 8- carbon acyl chloride were found highly effective (FIGs.2F-2G), while further modulation of tail length demonstrated lesser overall potency benefit. Notably, the top three SRB-lipidoids (1-8-10, 1-10-8, 1-12-6) in this library exhibited similar potency to MC3, all of which bear a total of 18 carbons in each tail (FIGs.2E-2H). Further increase or decrease in total carbon number of body and branch tails generally led to the loss of activity (FIG.2H). Without wishing to be bound by any theory, it is proposed herein that lipidoids certain exemplary lipidoids described herein comprising a total of 18 carbon atoms in the tail component (i.e., body tail and branch tail) represent optimal lipidoids. It has been reasoned that this number provides optimal hydrophobicity of SRB-lipidoids, which is closely correlated with alkyl chain length. This phenomenon is referred to herein as the “18-Carbon Rule”. That said, lipidoids comprising 18 carbon atoms in the tail component do not necessarily possess optimal efficacy for mRNA delivery in vivo (see 1-6-12) (FIG.2H), and compounds which lack 18 carbon atoms in the tail component are not definitively ineffective. It has been proposed that the symmetry of a branched tail / body tail could impact potency of the lipidoid. Therefore, a contour plot with both parameters (i.e., total carbon number and tail symmetry) was prepared to better visualize their influence on in vivo performance (FIG.2I; including symmetry formula). As expected, the potency of SRB- lipidoids was dependent not only on total carbon number, but also on the symmetry of body to branch tails. The symmetry of the lead 1-10-8 SRB-lipidoid was defined as 1 due to the relatively symmetrical structure of the body and branch tails, while the symmetries of the slightly less potent 1-8-10 and 1-12-6 SRB-lipidoids as well as the significantly less potent 1- 6-12 SRB-lipidoid were determined to be 1.7, 0.6, and 3, respectively. This finding suggests that, for SRB-lipidoids with a total carbon number of 18, less symmetry (i.e., more deviation - 74 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) from 1) is associated with less in vivo mRNA delivery potency. Taken together, these results indicate that the optimal tail region should follow “18-Carbon Rule” and have a symmetry of 1, which leads to the combination of a 10-carbon body tail and an 8-carbon branch tail. Next, the in vivo potency of purified SRB-lipidoids containing two branch tails was tested and compared to a SRB-lipidoid containing one branch tail (an intermediate metabolite of the standard SRB-lipidoid (FIGs.5A-5D). Purified 1-10-8 with two branch tails (referred to herein as “1-10-8(2)”) demonstrated comparable potency to the crude product, confirming that this structure is the active transfection ingredient ([M+H]+= 667.649 observed). In contrast, synthesized 1-10-8 with one branch tail (denoted as “1-10-8(1)”) was unable to encapsulate or deliver mRNA, highlighting the importance of both branch tails ([M+H]+= 541.557 observed). The degradability of SRB-lipidoids was then assessed. After treatment of 1-10-8(2) with esterase (an enzyme that hydrolyzes esters) for 1 h, its metabolite 1-10-8(1) was detected. Together, these results suggest that branch tails only endow transient potency for mRNA delivery, and their intracellular degradation is expected to cause the dissociation of LNPs and release of mRNA. Example 3: Optimizing the headgroup of SRB-lipidoids In the second stage of the optimization scheme described herein, the tail region was held constant at the optimal parameters identified above and the headgroup was varied to generate a second library of SRB-lipidoids (i.e., Library 2, FIG.6A and Tables.5-6). Table 5. Exemplary lipids (Library 2) Name Compound 2-10-8 (methylazanediyl)bis(decane-1,2-diyl) dioctanoate 3-10-8 (ethylazanediyl)bis(decane-1,2-diyl) dioctanoate 4-10-8 (2,2-dimethylhydrazine-1,1-diyl)bis(decane-1,2-diyl) dioctanoate - 75 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 5-10-8 ((4-methylpiperazin-1-yl)azanediyl)bis(decane-1,2-diyl) dioctanoate 6-10-8 ((2-methoxyethyl)azanediyl)bis(decane-1,2-diyl) dioctanoate 7-10-8 ((2-(dimethylamino)ethyl)azanediyl)bis(decane-1,2-diyl) dioctanoate 8-10-8 ((4-(dimethylamino)butyl)azanediyl)bis(decane-1,2-diyl) dioctanoate 9-10-8   ((2-(diethylamino)ethyl)azanediyl)bis(decane-1,2-diyl) dioctanoate 10-10-8 ((3-(diethylamino)propyl)azanediyl)bis(decane-1,2-diyl) dioctanoate 11-10-8 ((2-(pyrrolidin-1-yl)ethyl)azanediyl)bis(decane-1,2-diyl) dioctanoate 12-10-8 ((3-(pyrrolidin-1-yl)propyl)azanediyl)bis(decane-1,2-diyl) dioctanoate - 76 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 13-10-8 ((3-(1H-imidazol-1-yl)propyl)azanediyl)bis(decane-1,2-diyl) dioctanoate 14-10-8 ((2-(piperidin-1-yl)ethyl)azanediyl)bis(decane-1,2-diyl) dioctanoate 15-10-8 ((3-(4-methylpiperazin-1-yl)propyl)azanediyl)bis(decane-1,2-diyl) dioctanoate 16-10-8   (ethane-1,2-diylbis(methylazanediyl))bis(decane-1,2-diyl) dioctanoate 17-10-8 (propane-1,3-diylbis(methylazanediyl))bis(decane-1,2-diyl) dioctanoate 18-10-8 (ethane-1,2-diylbis(ethylazanediyl))bis(decane-1,2-diyl) dioctanoate 19-10-8 (((methylazanediyl)bis(propane-3,1-diyl))bis(methylazanediyl))bis(decane-1,2- diyl) dioctanoate 20-10-8 - 77 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) piperazine-1,4-diylbis(decane-1,2-diyl) dioctanoate Table 6. Observed m / z ratios for exemplary SRB-lipidoids Library SRB-lipidoid Calculated Observed 1 1-6-6 498.44 499.50 1 1-6-8 554.50 555.62 1 1-6-10 610.56 611.56 1 1-6-12 666.63 667.73 1 1-6-14 722.69 723.71 1 1-8-6 554.50 555.46 1 1-8-8 610.56 611.46 1 1-8-10 666.63 667.73 1 1-8-12 722.69 723.59 1 1-8-14 778.75 779.85 1 1-10-6 610.56 611.46 1 1-10-8 666.63 667.80 1 1-10-10 722.69 723.79 1 1-10-12 778.75 779.67 1 1-10-14 834.82 835.71 1 1-12-6 666.63 667.73 1 1-12-8 722.69 723.79 1 1-12-10 778.75 779.85 1 1-12-12 834.82 835.90 1 1-12-14 890.88 891.92 1 1-14-6 722.69 723.79 1 1-14-8 778.75 779.75 1 1-14-10 834.82 835.92 1 1-14-12 890.88 891.82 1 1-14-14 946.94 948.01 2 1-10-8 666.63 667.80 2 2-10-8 595.55 596.75 2 3-10-8 609.57 610.58 2 4-10-8 624.58 625.48 2 5-10-8 679.62 680.74 2 6-10-8 639.58 640.79 2 7-10-8 652.61 653.62 2 8-10-8 680.64 681.54 2 9-10-8 680.64 681.76 2 10-10-8 694.66 695.87 2 11-10-8 678.63 679.62 2 12-10-8 692.64 693.65 2 13-10-8 689.61 690.51 2 14-10-8 692.64 693.76 2 15-10-8 721.67 722.88 2 16-10-8 652.61 653.62 2 17-10-8 666.63 667.53 2 18-10-8 680.64 681.76 - 78 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 2 19-10-8 737.70 738.91 2 20-10-8 650.60 651.58 3 11-6-12 678.63 679.61 3 11-8-10 678.63 679.62 3 11-12-6 678.63 679.61 3 11-12-10 758.69 759.70 To maintain their structural similarity, only amines that can attach two body tails were chosen. Twenty (20) chemically diverse amines were tested, including monoamines, diamines, polyamines, and hydrazines (Table 5). SRB-lipidoids were denoted as X-10-8, wherein X stands for amine number identifier (Table 1). The in vitro and in vivo performance of SRB-lipidoids in Library 2 was determined using the same experimental conditions as Library 1 (FIGs.6A-6B and FIG.7). Again, these SRB-lipidoids demonstrated a poor correlation between their in vitro and in vivo potencies (FIG.6D). In this library, six SRB-lipidoids (i.e., comprising headgroup 1, 7, 9, 10, 11, or 12) exhibited comparable or superior in vivo performance compared to MC3, despite the fact that none of them outperformed MC3 in vitro (FIGs.6C-6D). Interestingly, the results from Libraries 1 and 2 demonstrate that although potent in vivo SRB-lipidoids do not necessarily perform well in vitro, all of them surpass a baseline mRNA transfection threshold (i.e., 10,000 units, FIG.2E and FIG.6D). The relationship between headgroup structure and in vivo potency was further investigated. Certain amines compounds with only one primary amine (e.g., 2, 3 and 6), two secondary amines (e.g., 16-20) or a hydrazine group (e.g., 4 and 5) were found to provide LNPs which possess lesser efficacy than LNPs prepared from other SRB-lipidoids described herein (FIG.6C). Effective amines (e.g., 1, 7, and 9-12) shared certain structural similarities (e.g., diamines with one primary amine and one tertiary amine spaced by two or three carbons. Notably, both the length of spacer (e.g., 1 and 7-8) and the form of tertiary amine (e.g., 9, 11, and 14) critically impacted in vivo performance. The optimal spacer length was identified to be two or three methylene units, while the ideal tertiary amine structure was determined to be dimethylamino, diethylamino, or pyrrolidinyl (FIG.6E). Without wishing to be bound by any theory, certain preferred structural criteria of potent SRB-lipidoids are proposed herein: (i) total carbon number of tail components = 18; (ii) symmetry = 1; (iii) diamines with one primary amine and one dimethylamino-, diethylamino- or pyrrolidinyl-based tertiary amine spaced by two or three carbons. - 79 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) The lead SRB-lipidoid 11-10-8, roughly 5-fold more potent than MC3, was identified and used for subsequent studies. Benefitting from two branch tails, the packing parameter (P) of this molecule was determined to be 4.1 according to molecular dynamics simulations (FIG. 6F), suggesting that 11-10-8 adopted a more cone-shape structure than MC3 (4.07 versus 2.11, FIG.8). Example 4: Predicting performance and optimizing formulation of SRB-LNPs To further validate the structural criteria identified herein for SRB-lipidoids, the capacity to predict in vivo performance of unidentified SRB-lipidoids was sought. As a case study, four amine 11-based SRB-lipidoids that either violated the “18-Carbon Rule” or defied the optimal symmetry of the tail region were prepared: 11-6-12 (total carbon number = 18, symmetry = 3), 11-8-10 (total carbon number = 18, symmetry = 1.7), 11-12-6 (total carbon number = 18, symmetry = 0.6) and 11-12-10 (total carbon number = 22, symmetry = 1). It was predicted that these SRB-lipidoids would have less in vivo mRNA transfection potency relative to 11-10-8. Indeed, the experimental results confirmed this prediction (FIG. 9A); all four SRB-lipidoids that breached the aforementioned design criteria were less potent than 11-10-8. Moreover, despite having the same total carbon number of 18, 11-8-10 and 11- 12-6 with more symmetry exhibited similar or higher potency compared to MC3, but 11-6-12 with less symmetry dramatically lost potency as expected. Therefore, the structural criteria are highly predictive of in vivo performance of SRB-lipidoids. Next, the component ratio of 11-10-8 LNP was varied and screened a series of well- established LNP formulations (F1-6, FIG.9B and Table 7). F1 was the formulation used in initial screening, which comprised of crude 11-10-8, while purified 11-10-8 was used in F2- F6. Crude 11-10-8 and purified 11-10-8 had similar in vivo potency when the same LNP formula was used (F1 versus F2, FIG.9B), which was in line with previous observation (FIG. 5D). Moreover, all 11-10-8 LNP formulations outperformed the benchmark MC3 LNP formulation except F4, which possessed a higher molar percentage of DMG-PEG compared to other formulations (FIG.9B and Table 7). The top-performing formulation F5 (11-10- 8 / DOPE / Cholesterol / DMG-PEG = 40:10:48.5:1.5), roughly 7-fold more potent than MC3, was chosen for further in vivo studies. Table 7. Exemplary LNP formulations LNP Components Molar Ratio Weight Ratio - 80 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) F1 11-10-8a / DOPE / Chol / DMG- 36.8:21:40.4:1.8 16:10:10:3 PEG F2 11-10-8 / DOPE / Chol / DMG-PEG 36.8:21:40.4:1.8 16:10:10:3 F3 11-10-8 / DOPE / Chol / DMG-PEG 30.6:17.5:50.4:1.5 16:10:15:3 F4 11-10-8 / DOPE / Chol / DMG-PEG 35:16:46.5:2.5 16:8:12:4.5 F5 11-10-8 / DOPE / Chol / DMG-PEG 40:10:48.5:1.5 16:4.4:11:2.4 F6 11-10-8 / DSPC / Chol / DMG-PEG 50:10:38.5:1.5 16:3.7:7:1.9 MC3 MC3 / DSPC / Chol / DMG-PEG 50:10:38.5:1.5 16:3.7:7:1.9 LNPacrude 11-10-8. Example 5: Characterization and in vitro studies of SRB-LNPs The physiochemical properties of optimized LNP 11-10-8 were characterized. Cryogenic electron microscopy (Cryo-EM) revealed that 11-10-8 LNP possessed a dense spherical structure with a multi-lamellar shell and an amorphous core (FIG.9C). The mRNA encapsulation efficiency (EE) of 11-10-8 LNP was determined to be ~95% (FIG.9D). The hydrodynamic size of 11-10-8 LNP was approximately 71.2 nm with a low polydispersity index (PDI = 0.087) and a neutral surface charge (ζ = -1.09 mV). The apparent pKa of 11-10- 8 LNP was determined to be 6.12 (FIG.10). Due to its ionization ability, 11-10-8 LNP exhibited good hemocompatibility at pH 7.4, but increased hemolysis at pH 6.0 (FIG.11). This ionization behavior is critical for reducing the toxicity of LNPs and enhancing membrane disruption in the acidic endosome. In HepG2 cells, 11-10-8 LNP showed dose-dependent mLuc delivery with minimal toxicity at doses ranging from 0 to 240 ng / well (FIGs.12A-12B). Moreover, 11-10-8 LNP exhibited dose-dependent delivery of GFP mRNA (FIGs.13A-13D), and could achieve nearly 100% transfection efficiency at dose as low as 150 ng / mL. Next, the cellular uptake routes of 11-10-8 LNP were examined. The endocytosis of 11-10-8 LNP was highly dependent on lipid rafts (FIG.9E), as pre-treatment with methyl-β cyclodextrin (Mβ-CD), an inhibitor of lipid raft-mediated endocytosis, completely suppressed mLuc delivery. Other internalization pathways (e.g., macropinocytosis, clathrin-mediated endocytosis and caveolae-mediated endocytosis) were also involved but contributed less to overall LNP internalization. - 81 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) Example 6: Hepatic delivery of mRNA-based therapeutics and gene editors using SRB- LNPs Hepatic delivery of mRNA-based therapeutics and gene editors holds great promise for protein supplementation therapies and genome editing therapies, respectively. Therefore, the potential of SRB-LNPs for these applications was further demonstrated by comparing 11- 10-8 LNP with MC3 LNP (Table 8). Ex vivo BLI results confirmed that 11-10-8 LNP predominantly transfected the liver, resulting in 4.8-fold higher mLuc expression in the liver than MC3 LNP (FIG.14A). These results were consistent with previous in vivo BLI results (FIG.9B). Similarly, 11-10-8 LNP also outperformed MC3 LNP in hepatic delivery of GFP mRNA (FIG.14B). Immunofluorescence staining results further confirmed that 11-10-8 LNP mediated greater GFP expression than MC3 LNP in major liver cells, including hepatocytes, Kupffer cells and liver sinusoidal endothelial cells (FIGs.22A-22C). Table 8. Characterization of MC3 LNP, aminoalcohol lipidoid-formulated LNPs, and DB- LNPs in Library 1aLNP Size (nm) PDI Zeta potential (mV) Encapsulation efficiency (%) 1-6 134.3 ± 5.5 0.14 -3.4 ± 0.7 19.0 ± 0.9 1-8 147.3 ± 6.8 0.20 -2.3 ± 0.9 20.7 ± 2.0 1-10 139.7 ± 4.7 0.16 -2.7 ± 1.0 24.4 ± 1.0 1-12 157.7 ± 6.7 0.18 -2.1 ± 0.8 29.9 ± 4.0 1-14 148.7 ± 8.3 0.17 -1.6 ± 1.0 32.8 ± 3.2 1-6-6 135.3 ± 5.5 0.21 -0.7 ± 1.3 32.6 ± 2.2 1-6-8 156.7 ± 4.6 0.20 1.9 ± 1.1 42.7 ± 0.6 1-6-10 129.7 ± 4.5 0.14 -1.4 ± 1.1 62.6 ± 5.9 1-6-12 146.3 ± 14.0 0.16 1.4 ± 0.1 77.8 ± 1.8 1-6-14 157.0 ± 5.0 0.14 -2.2 ± 0.7 75.7 ± 1.3 1-8-6 195.0 ± 12.8 0.20 0.03 ± 0.4 39.1 ± 0.4 1-8-8 138.7 ± 9.6 0.17 -1.6 ± 1.1 62.4 ± 4.9 1-8-10 140.7 ± 4.0 0.20 -0.8 ± 1.0 70.1 ± 1.2 1-8-12 174.7 ± 7.5 0.20 -1.6 ± 1.9 82.1 ± 1.0 1-8-14 130.0 ± 4.4 0.19 -1.6 ± 0.3 76.7 ± 2.3 1-10-6 160.0 ± 6.9 0.17 -2.0 ± 0.3 59.6 ± 3.5 1-10-8 150.7 ± 4.2 0.16 -0.03 ± 0.6 78.5 ± 0.4 1-10-10 154.0 ± 8.2 0.18 1.3 ± 1.1 79.8 ± 1.4 1-10-12 131.7 ± 5.9 0.19 -2.6 ± 0.7 77.2 ± 1.8 1-10-14 154.3 ± 6.7 0.17 0.47 ± 0.6 61.8 ± 1.4 1-12-6 130.3 ± 4.7 0.15 -2.7 ± 0.7 79.8 ± 4.3 1-12-8 136.0 ± 5.6 0.15 -2.6 ± 1.1 81.4 ± 0.9 1-12-10 162.3 ± 7.6 0.20 -2.0 ± 1.5 76.1 ± 2.0 1-12-12 198.7 ± 4.7 0.19 -0.67 ± 1.1 73.2 ± 1.8 1-12-14 214.0 ± 21.7 0.22 -1.6 ± 0.8 50.9 ± 2.9 1-14-6 137.0 ± 3.5 0.20 -0.6 ± 1.9 78.8 ± 0.5 1-14-8 150.3 ± 3.5 0.21 1.1 ± 0.8 72.5 ± 2.1 1-14-10 134.7 ± 4.2 0.18 -0.33 ± 0.9 76.6 ± 1.6 - 82 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 1-14-12 227.3 ± 23.5 0.22 1.5 ± 1.0 58.6 ± 4.9 1-14-14 203.7 ± 10.2 0.23 0.13 ± 0.3 42.7 ± 0.8 MC3 145.8 ± 8.1 0.17 -1.4 ± 0.3 73.5 ± 2.1aLNPs were formulated by pipette mixing. The hydrodynamic size and PDI were obtained by dynamic light scattering (DLS) measurement in neutral PBS. The zeta potential of LNPs was determined by laser-Doppler electrophoresis. The mRNA encapsulation efficiency was determined using a modified Quant-iT RiboGreen RNA assay. Data are presented as mean ± SD (n = 3 biologically independent samples). Table 9. Exemplary LNP formulationsaFormulation Recipe Molar ratio Weight ratio F1 11-10-8* / DOPE / Chol / DMG-PEG 36.8:21:40.4:1.8 16:10:10:3 F2 11-10-8 / DOPE / Chol / DMG-PEG 36.8:21:40.4:1.8 16:10:10:3 F3 11-10-8 / DOPE / Chol / DMG-PEG 30.6:17.5:50.4:1.5 16:10:15:3 F4 11-10-8 / DOPE / Chol / DMG-PEG 35:16:46.5:2.5 16:8:12:4.5 F5 11-10-8 / DOPE / Chol / DMG-PEG 40:10:48.5:1.5 16:4.4:11:2.4 F6 11-10-8 / DSPC / Chol / DMG-PEG 50:10:38.5:1.5 16:3.7:7:1.9 MC3 LNP MC3 / DSPC / Chol / DMG-PEG 50:10:38.5:1.5 16:3.7:7:1.9a11-10-8* indicates crude 11-10-8. The weight ratio of lipidoid:RNA was kept at 10:1. Table 10. Characterization of benchmark LNPs formulated by microfluidic mixingaLNP Molar ratio Size (nm) PDIZeta potentialEncapsulation (mV)efficiency (%) MC3 MC3 / DSPC / Chol / DMG- LNPPEG = 50:10:38.5:1.564.5 ± 3.7 0.063 -1.3 ± 0.3 92.7 ± 1.3SM-102 SM-102 / DSPC / Chol / DMG- LNPPEG = 50:10:38.5:170.1 ± 1.5 0.076 -0.5 ± 0.1 95.6 ± 2.4a.5The hydrodynamic size and PDI were obtained by dynamic light scattering (DLS) measurement in neutral PBS. The zeta potential of LNPs was determined by laser-Doppler electrophoresis. The mRNA encapsulation efficiency was determined using a modified Quant-iT RiboGreen RNA assay. Data are presented as mean ± SD (n = 3 biologically independent samples). Given the growing interest in CRISPR / Cas9-mediated gene editing as exemplified by Intellia’s LNP-mediated liver transthyretin (TTR) knockout, the potential of 11-10-8 LNP in this application was explored. A single injection of 11-10-8 LNP co-delivering Cas9 mRNA and TTR sgRNA at a clinically relevant dose (1 mg RNA / kg) led to ~30% insertions and deletions (indels) at the TTR locus and ~50% reduction of serum TTR (FIGs.14D-14E). In contrast, MC3 LNP only achieved ~6% indels and ~18% reduction of serum TTR. Notably, no observable hepatotoxicity was induced by both LNPs as revealed by normal levels of alanine transaminase (ALT) and aspartate aminotransferase (AST) in treated mice (FIGs. 15A-15B). Moreover, no abnormal innate immune responses were observed after examining 13 cytokines (Table 11). Together, these results strongly demonstrate great potential of the - 83 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) SRB-LNPs described herein in hepatic delivery of mRNA-based therapeutics and gene editors. Table 11. Innate immune responses after LNP treatmentaLNP Mouse No. IFN-γ CXCL1 TNF-α CCL2 IL-12p70 1 <0.86273 0.59578 <0.49209 <2.56921 <0.09701 Control 2 <0.86273 0.61765 <0.49209 <2.56921 <0.09701 3 <0.86273 0.83858 <0.49209 <2.56921 0.10067 11-10- 4 <0.86273 0.71050 <0.49209 <2.56921 <0.09701 8 5 <0.86273 0.62192 <0.49209 <2.56921 0.11071 LNP 6 <0.86273 0.60464 <0.49209 <2.56921 <0.09701 7 <0.86273 0.77333 <0.49209 <2.56921 <0.09701 MC3 8 <0.86273 1.01819 <0.49209 <2.56921 <0.09701 LNP 9 <0.86273 0.64890 <0.49209 <2.56921 <0.09701 LNP Mouse No. CCL5 IL-1β CXCL10 GM-CSF IL-10 1 <0.39291 0.27692 <1.84321 <0.36468 <0.44062 Control 2 <0.39291 <0.27548 <1.84321 <0.36468 0.677674 3 <0.39291 <0.27548 1.96991 <0.36468 0.484852 11-10- 4 0.54404 <0.27548 2.79788 <0.36468 0.613821 8 5 0.43782 0.28975 3.42005 <0.36468 0.524590 LNP 6 <0.39291 <0.27548 3.76349 <0.36468 0.524590 7 0.42437 <0.27548 5.56919 <0.36468 0.543476 MC3 8 0.71032 <0.27548 6.42526 <0.36468 0.579595 LNP 9 0.67270 <0.27548 2.97137 <0.36468 <0.44062 LNP Mouse No. IFN-β IFN-α IL-6 1 <0.41327 <0.38091 <0.29444 Control 2 <0.41327 <0.38091 <0.29444 3 <0.41327 <0.38091 <0.29444 11-10- 4 <0.41327 0.42651 <0.29444 8 5 <0.41327 0.38942 0.35976 LNP 6 <0.41327 0.50840 <0.29444 MC3 7 <0.41327 0.54182 0.76111 - 84 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) LNP 8 <0.41327 0.70700 <0.29444 9 <0.41327 0.54730 <0.29444an=3 biologically independent samples; results of cytokine concentrations (in pg / mL) are provided; “<” indicates the cytokine concentration is lower than the minimum detectable concentration. Cas9 mRNA / TTR sgRNA-loaded LNPs (1 mg / kg) were i.v. injected into mice and serum was collected at 24 h post treatment for cytokine analysis. All cytokines were either undetectable or at low levels (< 10 pg / mL) for both LNPs, which were comparable with the untreated control group. Example 7: Hepatic delivery of mRNA-based therapeutics using SRB-LNPs Hepatic delivery of mRNA-based therapeutics holds great promise for protein supplementation therapies. FGF21 is a pleiotropic metabolic hormone primarily secreted by the liver, which is a promising therapeutic agent for obesity, type 2 diabetes and non- alcoholic steatohepatitis. Next, 11-10-8 LNP was assessed for the delivery of a therapeutic mRNA encoding human FGF21. FGF21 is a metabolic hormone primarily secreted by liver, and its supplementation holds promise for the treatment of obesity, type 2 diabetes and non- alcoholic steatohepatitis. After 6 h post-administration of FGF21 mRNA-loaded 11-10-8 LNP, a high level of circulating FGF21 (50,991 ± 2,228 pg / mL) were detected in mice, which was 4.8-fold higher than that detected in MC3 LNP-treated mice (10,518 ± 2,683 pg / mL, FIG.14C). Moreover, the area under curve (AUC), a pharmacokinetic metric of therapeutic exposure, of FGF21 was 4.7-fold greater in 11-10-8 LNP-treated mice than that in MC3 LNP-treated mice (Table 12). Table 12. Serum FGF21 protein expression in mice during the time interval 0-48 h LNP 11-10-8 LNP MC3 LNP AUC (pg•h / mL) 849,378 ± 56,351 180,793 ± 67,275aAUC (area under curve); mice were i.v. injected with FGF21 mRNA-loaded LNPs at an mRNA dose of 0.25 mg / kg. Serum was collected at 0, 6, 30, and 48 h post-injection and analyzed by ELISA. Data are presented as mean ± SD (n = 4). Male mice were fed a HFD for three weeks to induce obesity (body weight ~30g) and fatty liver, and then i.v. injected with FGF21 mRNA-loaded LNPs. The expression of FGF21 peaked (58.8 ± 1.1 ng / mL) at 12 h post-administration of FGF21 mRNA-loaded 11-10-8 LNP, which was five-fold higher than that in FGF21 mRNA-loaded MC3 LNP-treated mice (11.8 ± 1.1 ng / mL, FIG.14C). Moreover, the area under curve (AUC) – a pharmacokinetic metric of therapeutic exposure – of FGF21 was 4.6-fold greater in 11-10-8 LNP-treated mice - 85 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) than that in MC3 LNP-treated mice (1104.1 ± 12.2 ng ^h / mL versus 240.4 ± 7.8 ng ^h / mL, FIG.14C). To further demonstrate the therapeutic potential of this FGF21 mRNA therapy, its weight-reducing and lipid-lowering effects were examined in obese mice. Obese mice were treated with various LNP formulations every other day for three doses (FIG.17A). While obese mice treated with PBS or mLuc-loaded LNPs gradually increased weight, obese mice treated with FGF21 mRNA-loaded 11-10-8 LNP or FGF21 mRNA-loaded MC3 LNP lost weight or maintained weight, respectively (FIG.17B). At the end of this experiment, body weight as well as liver weight of obese mice treated with FGF21 mRNA-loaded 11-10-8 LNP was significantly reduced compared to that of obese mice treated with other formulations (FIGs.17C-17D). Hematoxylin and eosin (H&E) staining of livers showed less vacuoles and reduced liver steatosis in the obese mice treated with FGF21 mRNA-loaded 11-10-8 LNP compared to those obese mice treated with other formulations (FIGs.17E-17F). It is worth mentioning that although MC3 LNP-based FGF21 mRNA therapy exhibited weight-reducing and lipid-lowering effects to some extent in obese mice, they were less obvious than 11-10-8 LNP-based therapy, presumably due to the less FGF21 expression (FIG.14C). Together, these results suggest that 11-10-8 LNP-based FGF21 mRNA therapy could be a promising approach for treating obesity and fatty liver. While 11-10-8 LNP was more effective at delivering large RNA constructs compared to MC3 LNP, it was unclear if the LNPs described herein possessed advantages in delivering small interference RNA (siRNA), for which the MC3 LNP is approved. Interestingly, when TTR siRNA was delivered, 11-10-8 LNP and MC3 LNP showed equivalent potency with the same median effective dose (ED50= 0.03 mg / kg; FIGs.16A-16B). Owing to the potent silencing effect, a single injection of 11-10-8 LNP at a clinically relevant dose (1 mg siRNA / kg) resulted in ~100% reduction of serum TTR on day 3 and nearly 40% reduction on day 28 (FIG.16B). Apart from systemic delivery, we further showed that 11-10-8 LNP mediated strong intramuscular (i.m.) mRNA delivery (FIG.23), which outperformed the benchmark SM-102 LNP that has been approved by FDA for mRNA vaccine delivery. Taken together, these results suggest that the SRB-LNPs described herein may represent a universal platform for robust delivery of various nucleic acid (e.g., RNA) constructs. Enumerated Embodiments The following exemplary embodiments are provided, the numbering of which is not - 86 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) to be construed as designating levels of importance: Embodiment 1 provides a compound formula (I), or a salt or solvate thereof: (I), wherein: A is selected from the group consisting of ; R1aand R1bare each ; R2is selected from the group 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-C8 heterocycloalkyl, and N(Ra)(Rb); R3aand R3bare each independently selected from the group consisting of optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C6heteroalkyl, optionally substituted C3-C6heteroalkenyl, and optionally substituted C2-C8 heterocycloalkyl; each occurrence of R4aand R4bis independently H; each occurrence of R5ais independently selected from the group consisting of optionally substituted C1-C24alkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C2-C24 heteroalkyl, optionally substituted C2-C12 heterocycloalkyl, optionally substituted C2-C24alkylenyl, optionally substituted C3-C24heteroalkylenyl, optionally substituted C2-C24 alkynylenyl, and optionally substituted C3-C24 heteroalkynylenyl; each occurrence of R5bis H; each occurrence of R6is independently selected from the group consisting of optionally substituted C1-C24alkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C2-C24 heteroalkyl, optionally substituted C2-C12 heterocycloalkyl, optionally substituted C2-C24alkylenyl, optionally substituted C3-C24heteroalkylenyl, optionally substituted C2-C24 alkynylenyl, and optionally substituted C3-C24 heteroalkynylenyl; L is selected from the group consisting of optionally substituted C2-C6alkylenyl and optionally substituted C2-C8 heteroalkylenyl; and Raand Rbare each independently selected from the group consisting of H and optionally substituted C1-C6alkyl. - 87 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) Embodiment 2 provides the compound of Embodiment 1, wherein each occurrence of optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynylenyl, optionally substituted cycloalkyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted alkynylenyl, optionally substituted C2-C8heterocycloalkyl, optionally substituted alkylenyl, and optionally substituted heteroalkylenyl is independently optionally substituted with at least one substituent selected from the group consisting of C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 haloalkyl, C1-C3 haloalkoxy, phenoxy, halogen, CN, NO2, OH, N(R’)(R’’), C(=O)R’, C(=O)OR’, OC(=O)OR’, C(=O)N(R’)(R’’), S(=O)2N(R’)(R’’), N(R’)C(=O)R’’, N(R’)S(=O)2R’’, C2-C8 heteroaryl, and phenyl optionally substituted with at least one halogen, wherein each occurrence of R’ and R’’ is independently selected from the group consisting of H, C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 haloalkyl, benzyl, and phenyl, or wherein R’ and R’’ can combine with the nitrogen atom to which they are bound to form a C2-C8 heterocycloalkyl (e.g., cyclic tertiary amine). Embodiment 3 provides the compound of Embodiment 1 or 2, wherein R5ais selected from the group consisting of ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, and tetradecyl. Embodiment 4 provides the compound of any one of Embodiments 1-3, wherein R5ais selected from the group consisting of n-propyl, n-pentyl, n-heptyl, n-nonyl, n-undecyl, and n-tridecyl. Embodiment 5 provides the compound of any one of Embodiments 1-4, wherein R6is selected from the group consisting of ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, and tetradecyl. Embodiment 6 provides the compound of any one of Embodiments 1-5, wherein R6is selected from the group consisting of n-butyl, n-hexyl, n-octyl, n-decyl, and n-dodecyl. Embodiment 7 provides the compound of any one of Embodiments 1-6, wherein R1a - 88 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) , 7, wherein R5a 18 carbon atoms. Embodiment 9 provides the compound of any one of Embodiments 1-8, wherein R2is a C2-C3alkyl substituted with at least one cyclic tertiary amine or acyclic tertiary amine. Embodiment 10 provides the compound of any one of Embodiments 1-9, wherein R2, L is selected from the group consisting of -(CH2)2-, -(CH2)3-, and -(CH2)3N(Me)(CH2)3-. Embodiment 12 provides the compound of any one of Embodiments 1-8 and 11, wherein one of the following applies: (a) R3aand R3bare each independently selected from the group consisting of Me and Et; or (b) R3aand R3bcombine with the nitrogen atoms to which they are bound to form , optionally wherein L is -(CH2)2-. the compound of any one of Embodiments 1-12, wherein A , , - 89 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) , which is group ((3-(dimethylamino)propyl)azanediyl)bis(hexane-1,2-diyl) dihexanoate; ((3-(dimethylamino)propyl)azanediyl)bis(hexane-1,2-diyl) dioctanoate; ((3-(dimethylamino)propyl)azanediyl)bis(hexane-1,2-diyl) bis(decanoate); ((3-(dimethylamino)propyl)azanediyl)bis(hexane-1,2-diyl) didodecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(hexane-1,2-diyl) ditetradecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(octane-1,2-diyl) dihexanoate; ((3-(dimethylamino)propyl)azanediyl)bis(octane-1,2-diyl) dioctanoate; ((3-(dimethylamino)propyl)azanediyl)bis(octane-1,2-diyl) bis(decanoate); ((3-(dimethylamino)propyl)azanediyl)bis(octane-1,2-diyl) didodecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(octane-1,2-diyl) ditetradecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(decane-1,2-diyl) dihexanoate; ((3-(dimethylamino)propyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((3-(dimethylamino)propyl)azanediyl)bis(decane-1,2-diyl) bis(decanoate); ((3-(dimethylamino)propyl)azanediyl)bis(decane-1,2-diyl) didodecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(decane-1,2-diyl) ditetradecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(dodecane-1,2-diyl) dihexanoate; ((3-(dimethylamino)propyl)azanediyl)bis(dodecane-1,2-diyl) dioctanoate; ((3-(dimethylamino)propyl)azanediyl)bis(dodecane-1,2-diyl) bis(decanoate); ((3-(dimethylamino)propyl)azanediyl)bis(dodecane-1,2-diyl) didodecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(dodecane-1,2-diyl) ditetradecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(tetradecane-1,2-diyl) dihexanoate; ((3-(dimethylamino)propyl)azanediyl)bis(tetradecane-1,2-diyl) dioctanoate; ((3-(dimethylamino)propyl)azanediyl)bis(tetradecane-1,2-diyl) bis(decanoate); ((3-(dimethylamino)propyl)azanediyl)bis(tetradecane-1,2-diyl) didodecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(tetradecane-1,2-diyl) ditetradecanoate; (methylazanediyl)bis(decane-1,2-diyl) dioctanoate; (ethylazanediyl)bis(decane-1,2-diyl) dioctanoate; - 90 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) (2,2-dimethylhydrazine-1,1-diyl)bis(decane-1,2-diyl) dioctanoate; ((4-methylpiperazin-1-yl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((2-methoxyethyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((2-(dimethylamino)ethyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((4-(dimethylamino)butyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((2-(diethylamino)ethyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((3-(diethylamino)propyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((2-(pyrrolidin-1-yl)ethyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((3-(pyrrolidin-1-yl)propyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((3-(1H-imidazol-1-yl)propyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((2-(piperidin-1-yl)ethyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((3-(4-methylpiperazin-1-yl)propyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; (ethane-1,2-diylbis(methylazanediyl))bis(decane-1,2-diyl) dioctanoate; (propane-1,3-diylbis(methylazanediyl))bis(decane-1,2-diyl) dioctanoate; (ethane-1,2-diylbis(ethylazanediyl))bis(decane-1,2-diyl) dioctanoate; (((methylazanediyl)bis(propane-3,1-diyl))bis(methylazanediyl))bis(decane-1,2-diyl) dioctanoate; and piperazine-1,4-diylbis(decane-1,2-diyl) dioctanoate. Embodiment 15 provides a lipid nanoparticle (LNP) composition comprising: (a) at least one ionizable lipid compound of any one of Embodiments 1-14; (b) at least one helper lipid; (c) at least one cholesterol lipid; and (d) at least one polymer-conjugated lipid and / or a modified derivative thereof. Embodiment 16 provides the LNP of Embodiment 15, wherein the at least one ionizable lipid comprises about 30 mol% to about 60 mol% of the LNP. Embodiment 17 provides the LNP of Embodiment 15 or 16, wherein the at least one ionizable lipid comprises about 30.6 mol%, 35.0 mol%, 36.8 mol%, 40 mol%, or about 50 mol% of the LNP. Embodiment 18 provides the LNP of any one of Embodiments 15-17, wherein the at least one ionizable lipid comprises ((2-(pyrrolidin-1-yl)ethyl)azanediyl)bis(decane-1,2-diyl) dioctanoate. Embodiment 19 provides the LNP of any one of Embodiments 15-18, wherein the at least one helper lipid comprises about 1 mol% to about 30 mol% of the LNP. Embodiment 20 provides the LNP of any one of Embodiments 15-19, wherein the at - 91 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) least one helper lipid comprises about 10 mol%, 16 mol%, 17.5 mol%, or about 21 mol% of the LNP. Embodiment 21 provides the LNP of any one of Embodiments 15-20, wherein the at least one helper lipid comprises dioleoylphosphatidylethanolamine (DOPE). Embodiment 22 provides the LNP of any one of Embodiments 15-21, wherein the cholesterol comprises about 30 mol% to about 60 mol% of the LNP. Embodiment 23 provides the LNP of any one of Embodiments 15-22, wherein the cholesterol comprises about 38.5 mol%, 40.4 mol%, 46.5 mol%, 48.5 mol%, or about 50.4 mol% of the LNP. Embodiment 24 provides the LNP of any one of Embodiments 15-23, wherein the at least one polymer-conjugated lipid and / or a modified derivative thereof comprises about 0.1 mol% to about 10 mol% of the LNP. Embodiment 25 provides the LNP of any one of Embodiments 15-24, wherein the at least one polymer-conjugated lipid and / or a modified derivative thereof comprises about 1.5 mol%, 1.8 mol%, or about 2.5 mol% of the LNP. Embodiment 26 provides the LNP of any one of Embodiments 15-25, wherein the at least one polymer-conjugated lipid and / or a modified derivative thereof comprises 1,2- dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (DMG-PEG-2000). Embodiment 27 provides the LNP of any one of Embodiments 15-26, wherein the LNP has a molar ratio of (a):(b):(c):(d) which is selected from the group consisting of about 36.8:21:40.4:1.8, 30.6:17.5:50.4:1.5, 35:16:46.5:2.5, 40:10:48.5:1.5, and about 50:10:38.5:1.5. Embodiment 28 provides the LNP of any one of Embodiments 15-27, wherein the LNP has a weight ratio of (a):(b):(c):(d) which is selected from the group consisting of about 16:10:10:3, 16:10:15:3, 16:8:12:4.5, 16:4.4:11:2.4, and about 16:3.7:7:1.9. Embodiment 29 provides the LNP of any one of Embodiments 15-28, wherein the LNP further comprises at least one 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 comprises a nucleic acid. Embodiment 32 provides the LNP of Embodiment 31, wherein the nucleic acid cargo is DNA or RNA. - 92 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) 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 any one of Embodiments 29-33, wherein the cargo is at least partially encapsulated in the LNP. Embodiment 35 provides the LNP of any one of Embodiments 29-34, wherein the ionizable lipid and nucleic acid cargo have a weight ratio ranging from about 20:1 to about 5:1 (ionizable lipid:cargo (w / w)), optionally wherein the weight ratio is about 10:1. Embodiment 36 provides the LNP of any one of Embodiments 32-35, wherein the nucleic acid cargo comprises mRNA, optionally wherein the nucleic acid cargo further comprises sgRNA. Embodiment 37 provides the LNP of Embodiment 36, wherein the mRNA comprises Cas9 mRNA and the sRNA comprises TTR sgRNA. Embodiment 38 provides a method of preparing the ionizable lipid compound of formula (I) of any one of Embodiments 1-14: (I), the method comprising: (a) contacting an amine compound of formula (A1) or (A2): at least , to form an alkoxide (b) contacting the alkoxide intermediate and an activated acid compound of formula (C): , wherein X is a Embodiment 39 provides the method of Embodiment 38, wherein at least one of the following applies: (a) the amine compound and epoxide have a molar ratio of about 1:2.4 (ratio of (A1) and (A2):(B)); - 93 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) (b) the contacting in step (a) occurs at a temperature of about 80 °C; (c) the contacting in step (a) occurs for a period of about 48 h; (d) the alkoxide intermediate is dissolved in a solvent before contacting in step (b), optionally wherein the solvent is dichloromethane; (e) the contacting in step (b) occurs in the present of a base, optionally wherein the base is a trialkylamine, optionally wherein the trialkylamine is triethylamine; and (f) X is Cl. Embodiment 40 provides the method of Embodiment 38 or 39, wherein the amine is selected from the group consisting of N1,N1-dimethylpropane-1,3-diamine, methylamine, ethylamine, 1,1-dimethylhydrazine, 2-methoxyethan-1amine, 2-methoxyethan-1-amine, 2- methoxyethan-1-amine, N1,N1-dimethylethane-1,2-diamine, N1,N1-dimethylbutane-1,4- diamine, N1,N1-diethylethane-1,2-diamine, N1,N1-diethylpropane-1,3-diamine, 2-(pyrrolidin- 1-yl)ethan-1-amine, 3-(pyrrolidin-1-yl)propan-1-amine, 3-(1H-imidazol-1-yl)propan-1- amine, 2-(piperidin-1-yl)ethan-1-amine, 3-(4-methylpiperazin-1-yl)propan-1-amine, N1,N2- dimethylethane-1,2-diamine, N1,N3-dimethylpropane-1,3-diamine, N1,N2-diethylethane-1,2- diamine, N1,N3-dimethyl-N1-(3-(methylamino)propyl)propane-1,3-diamine, and piperazine. Embodiment 41 provides the method of any one of Embodiments 38-40, wherein the epoxide is selected from the group consisting of 2-butyloxirane, 2-pentyloxirane, 2- hexyloxirane, 2-heptyloxirane, 2-octyloxirane, 2-nonyloxirane, 2-decyloxirane, 2- undecyloxirane, and 2-dodecyloxirane. Embodiment 42 provides the method of any one of Embodiments 38-41, wherein the activated acid is selected from the group consisting of hexanoyl chloride, heptanoyl chloride, octanoyl chloride, nonanoyl chloride, decanoyl chloride, undecanoyl chloride, dodecanoyl chloride, tridecanoyl chloride, and tetradecanoyl chloride. Embodiment 43 provides a pharmaceutical composition comprising the lipid nanoparticle of any one of Embodiments 15-37 and at least one pharmaceutically acceptable carrier. Embodiment 44 provides the pharmaceutical composition of Embodiment 43, wherein the composition further comprises at least one adjuvant. Embodiment 45 provides a method of treating, preventing, and / or ameliorating a disease or disorder in a subject, the method comprising administering to the subject at least one lipid nanoparticle (LNP) of any one of Embodiments 29-37. Embodiment 46 provides the method of Embodiment 45, wherein the LNP is - 94 - 52985306.3 Attorney Docket No.046483-7436WO1(03708) formulated as a pharmaceutical composition comprising at least one pharmaceutically acceptable carrier. Embodiment 47 provides the method of Embodiment 45 or 46, wherein the disease is cancer. Embodiment 48 provides the method of Embodiment 47, wherein the cancer is at least one selected from the group consisting of pancreatic cancer, colorectal cancer, bladder cancer, breast cancer, prostate cancer, renal cancer, hepatocellular cancer, lung cancer, ovarian cancer, cervical cancer, gastric cancer, esophageal cancer, head and neck cancer, melanoma, neuroendocrine cancer, CNS cancer, brain cancer, bone cancer, soft tissue sarcoma, non-small cell lung cancer, small-cell lung cancer, or colon cancer. Embodiment 49 provides the method of Embodiment 47 or 48, wherein the subject is further administered at least one additional agent or therapy useful for treating, preventing, and / or ameliorating cancer in the subject. Embodiment 50 provides the method of any one of Embodiments 45-49, wherein the subject is a mammal. Embodiment 51 provides the method of Embodiment 50, wherein the mammal is a human. 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. - 95 - 52985306.3

Claims

Attorney Docket No.046483-7436WO1(03708) CLAIMS What is claimed is:

1. A compound formula (I), or a salt or solvate thereof: (I), wherein: A is selected from the group consisting ;R1aand R1bare each ; R2is selected from the group substituted C1-C6 alkyl,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); R3aand R3bare each independently selected from the group consisting of optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C6heteroalkyl, optionally substituted C3-C6heteroalkenyl, and optionally substituted C2-C8 heterocycloalkyl; each occurrence of R4aand R4bis independently H; each occurrence of R5ais independently selected from the group consisting of optionally substituted C1-C24alkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C2-C24 heteroalkyl, optionally substituted C2-C12 heterocycloalkyl, optionally substituted C2-C24alkylenyl, optionally substituted C3-C24heteroalkylenyl, optionally substituted C2-C24 alkynylenyl, and optionally substituted C3-C24 heteroalkynylenyl; each occurrence of R5bis H; each occurrence of R6is independently selected from the group consisting of optionally substituted C1-C24alkyl, optionally substituted C3-C12cycloalkyl, optionally substituted C2-C24 heteroalkyl, optionally substituted C2-C12 heterocycloalkyl, optionally substituted C2-C24alkylenyl, optionally substituted C3-C24heteroalkylenyl, optionally substituted C2-C24 alkynylenyl, and optionally substituted C3-C24 heteroalkynylenyl; L is selected from the group consisting of optionally substituted C2-C6 alkylenyl and optionally substituted C2-C8heteroalkylenyl; and - 96 - 52985306.3Attorney Docket No.046483-7436WO1(03708) Raand Rbare each independently selected from the group consisting of H and optionally substituted C1-C6alkyl.

2. The compound of claim 1, wherein each occurrence of optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynylenyl, optionally substituted cycloalkyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted alkynylenyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted alkylenyl, and optionally substituted heteroalkylenyl is independently optionally substituted with at least one substituent selected from the group consisting of C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6haloalkyl, C1-C3haloalkoxy, 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-C8heteroaryl, 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-C6alkyl, C3-C8cycloalkyl, C1-C6haloalkyl, benzyl, and phenyl, or wherein R’ and R’’ can combine with the nitrogen atom to which they are bound to form a C2-C8 heterocycloalkyl (e.g., cyclic tertiary amine).

3. The compound of claim 1 or 2, wherein R5ais selected from the group consisting of ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, and tetradecyl.

4. The compound of any one of claims 1-3, wherein R5ais selected from the group consisting of n-propyl, n-pentyl, n-heptyl, n-nonyl, n-undecyl, and n-tridecyl.

5. The compound of any one of claims 1-4, wherein R6is selected from the group consisting of ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, and tetradecyl.

6. The compound of any one of claims 1-5, wherein R6is selected from the group consisting of n-butyl, n-hexyl, n-octyl, n-decyl, and n-dodecyl.

7. The compound of any one of claims 1-6, wherein R1aand R1bare each independently - 97 - 52985306.3Attorney Docket No.046483-7436WO1(03708) selected from the group consisting of ,any one are R1aand R1beach independently comprise about 18 carbon atoms.

9. The compound of any one of claims 1-8, wherein R2is a C2-C3 alkyl substituted with at least one cyclic tertiary amine or acyclic tertiary amine.

10. The compound of any one of claims 1-9, wherein R2is selected from the group ,11. The compound of any one of claims 1-8, wherein L is selected from the group consisting of -(CH2)2-, -(CH2)3-, and -(CH2)3N(Me)(CH2)3-.

12. The compound of any one of claims 1-8 and 11, wherein one of the following applies: (a) R3aand R3bare each independently selected from the group consisting of Me and Et; or (b) R3aand R3bcombine with the nitrogen atoms to which they are bound to form - 98 - 52985306.3Attorney Docket No.046483-7436WO1(03708) , optionally wherein L is -(CH2)2-.

13. The any one of claims 1-12, wherein A is selected from the group , .

14. The compound of any one of claims 1-13, which is selected from the group consisting of: ((3-(dimethylamino)propyl)azanediyl)bis(hexane-1,2-diyl) dihexanoate; ((3-(dimethylamino)propyl)azanediyl)bis(hexane-1,2-diyl) dioctanoate; ((3-(dimethylamino)propyl)azanediyl)bis(hexane-1,2-diyl) bis(decanoate); ((3-(dimethylamino)propyl)azanediyl)bis(hexane-1,2-diyl) didodecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(hexane-1,2-diyl) ditetradecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(octane-1,2-diyl) dihexanoate; ((3-(dimethylamino)propyl)azanediyl)bis(octane-1,2-diyl) dioctanoate; ((3-(dimethylamino)propyl)azanediyl)bis(octane-1,2-diyl) bis(decanoate); ((3-(dimethylamino)propyl)azanediyl)bis(octane-1,2-diyl) didodecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(octane-1,2-diyl) ditetradecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(decane-1,2-diyl) dihexanoate; ((3-(dimethylamino)propyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((3-(dimethylamino)propyl)azanediyl)bis(decane-1,2-diyl) bis(decanoate); ((3-(dimethylamino)propyl)azanediyl)bis(decane-1,2-diyl) didodecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(decane-1,2-diyl) ditetradecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(dodecane-1,2-diyl) dihexanoate; - 99 - 52985306.3Attorney Docket No.046483-7436WO1(03708) ((3-(dimethylamino)propyl)azanediyl)bis(dodecane-1,2-diyl) dioctanoate; ((3-(dimethylamino)propyl)azanediyl)bis(dodecane-1,2-diyl) bis(decanoate); ((3-(dimethylamino)propyl)azanediyl)bis(dodecane-1,2-diyl) didodecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(dodecane-1,2-diyl) ditetradecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(tetradecane-1,2-diyl) dihexanoate; ((3-(dimethylamino)propyl)azanediyl)bis(tetradecane-1,2-diyl) dioctanoate; ((3-(dimethylamino)propyl)azanediyl)bis(tetradecane-1,2-diyl) bis(decanoate); ((3-(dimethylamino)propyl)azanediyl)bis(tetradecane-1,2-diyl) didodecanoate; ((3-(dimethylamino)propyl)azanediyl)bis(tetradecane-1,2-diyl) ditetradecanoate; (methylazanediyl)bis(decane-1,2-diyl) dioctanoate; (ethylazanediyl)bis(decane-1,2-diyl) dioctanoate; (2,2-dimethylhydrazine-1,1-diyl)bis(decane-1,2-diyl) dioctanoate; ((4-methylpiperazin-1-yl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((2-methoxyethyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((2-(dimethylamino)ethyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((4-(dimethylamino)butyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((2-(diethylamino)ethyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((3-(diethylamino)propyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((2-(pyrrolidin-1-yl)ethyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((3-(pyrrolidin-1-yl)propyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((3-(1H-imidazol-1-yl)propyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((2-(piperidin-1-yl)ethyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; ((3-(4-methylpiperazin-1-yl)propyl)azanediyl)bis(decane-1,2-diyl) dioctanoate; (ethane-1,2-diylbis(methylazanediyl))bis(decane-1,2-diyl) dioctanoate; (propane-1,3-diylbis(methylazanediyl))bis(decane-1,2-diyl) dioctanoate; (ethane-1,2-diylbis(ethylazanediyl))bis(decane-1,2-diyl) dioctanoate; (((methylazanediyl)bis(propane-3,1-diyl))bis(methylazanediyl))bis(decane-1,2-diyl) dioctanoate; and piperazine-1,4-diylbis(decane-1,2-diyl) dioctanoate.

15. A lipid nanoparticle (LNP) composition comprising: (a) at least one ionizable lipid compound of any one of claims 1-14; (b) at least one helper lipid; (c) at least one cholesterol lipid; and - 100 - 52985306.3Attorney Docket No.046483-7436WO1(03708) (d) at least one polymer-conjugated lipid and / or a modified derivative thereof.

16. The LNP of claim 15, wherein the at least one ionizable lipid comprises about 30 mol% to about 60 mol% of the LNP.

17. The LNP of claim 15 or 16, wherein the at least one ionizable lipid comprises about 30.6 mol%, 35.0 mol%, 36.8 mol%, 40 mol%, or about 50 mol% of the LNP.

18. The LNP of any one of claims 15-17, wherein the at least one ionizable lipid comprises ((2-(pyrrolidin-1-yl)ethyl)azanediyl)bis(decane-1,2-diyl) dioctanoate.

19. The LNP of any one of claims 15-18, wherein the at least one helper lipid comprises about 1 mol% to about 30 mol% of the LNP.

20. The LNP of any one of claims 15-19, wherein the at least one helper lipid comprises about 10 mol%, 16 mol%, 17.5 mol%, or about 21 mol% of the LNP.

21. The LNP of any one of claims 15-20, wherein the at least one helper lipid comprises dioleoylphosphatidylethanolamine (DOPE).

22. The LNP of any one of claims 15-21, wherein the cholesterol comprises about 30 mol% to about 60 mol% of the LNP.

23. The LNP of any one of claims 15-22, wherein the cholesterol comprises about 38.5 mol%, 40.4 mol%, 46.5 mol%, 48.5 mol%, or about 50.4 mol% of the LNP.

24. The LNP of any one of claims 15-23, wherein the at least one polymer-conjugated lipid and / or a modified derivative thereof comprises about 0.1 mol% to about 10 mol% of the LNP.

25. The LNP of any one of claims 15-24, wherein the at least one polymer-conjugated lipid and / or a modified derivative thereof comprises about 1.5 mol%, 1.8 mol%, or about 2.5 mol% of the LNP. - 101 - 52985306.3Attorney Docket No.046483-7436WO1(03708) 26. The LNP of any one of claims 15-25, wherein the at least one polymer-conjugated lipid and / or a modified derivative thereof comprises 1,2-dimyristoyl-rac-glycero-3- methoxypolyethylene glycol-2000 (DMG-PEG-2000).

27. The LNP of any one of claims 15-26, wherein the LNP has a molar ratio of (a):(b):(c):(d) which is selected from the group consisting of about 36.8:21:40.4:1.8, 30.6:17.5:50.4:1.5, 35:16:46.5:2.5, 40:10:48.5:1.5, and about 50:10:38.5:1.

5.

28. The LNP of any one of claims 15-27, wherein the LNP has a weight ratio of (a):(b):(c):(d) which is selected from the group consisting of about 16:10:10:3, 16:10:15:3, 16:8:12:4.5, 16:4.4:11:2.4, and about 16:3.7:7:1.

9.

29. The LNP of any one of claims 15-28, wherein the LNP further comprises at least one 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 comprises a nucleic acid.

32. The LNP of claim 31, wherein the nucleic acid cargo 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 any one of claims 29-33, wherein the cargo is at least partially encapsulated in the LNP.

35. The LNP of any one of claims 29-34, wherein the ionizable lipid and nucleic acid cargo have a weight ratio ranging from about 20:1 to about 5:1 (ionizable lipid:cargo (w / w)), optionally wherein the weight ratio is about 10:

1. - 102 - 52985306.3Attorney Docket No.046483-7436WO1(03708) 36. The LNP of any one of claims 32-35, wherein the nucleic acid cargo comprises mRNA, optionally wherein the nucleic acid cargo further comprises sgRNA.

37. The LNP of claim 36, wherein the mRNA comprises Cas9 mRNA and the sRNA comprises TTR sgRNA.

38. A method of preparing the ionizable lipid compound of formula (I) of any one of claims 1-14: (I), the method comprising:(a) contacting an amine (A1) or (A2): at least, to form an alkoxide(b) contacting the alkoxide intermediate and an activated acid compound of formula (C): , wherein X is a39. The method of claim 38, wherein at least one of the following applies: (a) the amine compound and epoxide have a molar ratio of about 1:2.4 (ratio of (A1) and (A2):(B)); (b) the contacting in step (a) occurs at a temperature of about 80 °C; (c) the contacting in step (a) occurs for a period of about 48 h; (d) the alkoxide intermediate is dissolved in a solvent before contacting in step (b), optionally wherein the solvent is dichloromethane; (e) the contacting in step (b) occurs in the present of a base, optionally wherein the base is a trialkylamine, optionally wherein the trialkylamine is triethylamine; and - 103 - 52985306.3Attorney Docket No.046483-7436WO1(03708) (f) X is Cl.

40. The method of claim 38 or 39, wherein the amine is selected from the group consisting of N1,N1-dimethylpropane-1,3-diamine, methylamine, ethylamine, 1,1- dimethylhydrazine, 2-methoxyethan-1amine, 2-methoxyethan-1-amine, 2-methoxyethan-1- amine, N1,N1-dimethylethane-1,2-diamine, N1,N1-dimethylbutane-1,4-diamine, N1,N1- diethylethane-1,2-diamine, N1,N1-diethylpropane-1,3-diamine, 2-(pyrrolidin-1-yl)ethan-1- amine, 3-(pyrrolidin-1-yl)propan-1-amine, 3-(1H-imidazol-1-yl)propan-1-amine, 2- (piperidin-1-yl)ethan-1-amine, 3-(4-methylpiperazin-1-yl)propan-1-amine, N1,N2- dimethylethane-1,2-diamine, N1,N3-dimethylpropane-1,3-diamine, N1,N2-diethylethane-1,2- diamine, N1,N3-dimethyl-N1-(3-(methylamino)propyl)propane-1,3-diamine, and piperazine.

41. The method of any one of claims 38-40, wherein the epoxide is selected from the group consisting of 2-butyloxirane, 2-pentyloxirane, 2-hexyloxirane, 2-heptyloxirane, 2- octyloxirane, 2-nonyloxirane, 2-decyloxirane, 2-undecyloxirane, and 2-dodecyloxirane.

42. The method of any one of claims 38-41, wherein the activated acid is selected from the group consisting of hexanoyl chloride, heptanoyl chloride, octanoyl chloride, nonanoyl chloride, decanoyl chloride, undecanoyl chloride, dodecanoyl chloride, tridecanoyl chloride, and tetradecanoyl chloride.

43. A pharmaceutical composition comprising the lipid nanoparticle of any one of claims 15-37 and at least one pharmaceutically acceptable carrier.

44. The pharmaceutical composition of claim 43, wherein the composition further comprises at least one adjuvant.

45. A method of treating, preventing, and / or ameliorating a disease or disorder in a subject, the method comprising administering to the subject at least one lipid nanoparticle (LNP) of any one of claims 29-37.

46. The method of claim 45, wherein the LNP is formulated as a pharmaceutical composition comprising at least one pharmaceutically acceptable carrier. - 104 - 52985306.3Attorney Docket No.046483-7436WO1(03708) 47. The method of claim 45 or 46, wherein the disease is cancer.

48. The method of claim 47, wherein the cancer is at least one selected from the group consisting of pancreatic cancer, colorectal cancer, bladder cancer, breast cancer, prostate cancer, renal cancer, hepatocellular cancer, lung cancer, ovarian cancer, cervical cancer, gastric cancer, esophageal cancer, head and neck cancer, melanoma, neuroendocrine cancer, CNS cancer, brain cancer, bone cancer, soft tissue sarcoma, non-small cell lung cancer, small-cell lung cancer, or colon cancer.

49. The method of claim 47 or 48, wherein the subject is further administered at least one additional agent or therapy useful for treating, preventing, and / or ameliorating cancer in the subject.

50. The method of any one of claims 45-49, wherein the subject is a mammal.

51. The method of claim 50, wherein the mammal is a human. - 105 - 52985306.3

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