EP4731632A1 - Toll-like receptor (tlr) agonist lipidoid compounds, lipid nanoparticles (lnps) comprising the same, and methods of use thereof - Google Patents

Abstract

The present disclosure relates to lipidoid compounds comprising toll-like receptor (TLR) agonists, lipid nanoparticles (LNPs) comprising the same, and methods of use thereof. In certain embodiments, the LNPs described herein are useful for enhancing the therapeutic and/or prophylactic effect of vaccine compositions.

Description

Attorney Docket No.046483-7434WO1(03633) TITLE OF THE INVENTION Toll-Like Receptor (TLR) Agonist Lipidoid Compounds, Lipid Nanoparticles (LNPs) Comprising the Same, and Methods of Use Thereof CROSS-REFERENCE TO RELATED APPLICATIONS The present application claims priority under 35 U.S.C. § 119(e) to U.S. Provisional Patent Application No.63/509,452 filed June 21, 2023, the content of which is hereby incorporated by reference in its entirety herein. STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT This invention was made with government support under TR002776 awarded by the National Institutes of Health. The government has certain rights in the invention. BACKGROUND The coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has resulted in a global health crisis and millions of casualties. Vaccines have proven a valuable tool to reduce morbidity and mortality of this infectious disease. Currently, several vaccine modalities, including mRNA vaccines, antigen- expressing adenovirus, inactivated virus, and subunit vaccines, are either approved or under clinical development. Among these, mRNA vaccines not only demonstrate superior ability to elicit both neutralizing antibody (NAb) and T cell responses against multiple SARS-CoV-2 variants, but also offer low production cost with a short development and manufacturing period. Although mRNA was discovered in 1961, due to its susceptibility to enzymatic degradation, inefficient in vivo delivery, and high innate immunogenicity, its vaccine development has long been hampered until recently. The first two challenges are overcome by the incorporation of mRNA into a delivery system that can protect the mRNA from degradation and escort the mRNA across various biological barriers. Notably, four- component lipid nanoparticles (LNPs) comprising ionizable lipid (or lipidoid), phospholipid, PEGylated lipid, and cholesterol are the most clinically advanced mRNA delivery platform as exemplified by Pfizer/BioNTech’s BNT162b2 and Moderna’s mRNA-1273 vaccines. The last challenge is addressed through the incorporation of naturally occurring nucleosides such as 1-methylpseudouridine (m1ψ) into the in vitro transcribed mRNA sequence to avoid its - 1 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) innate immune recognition by pattern recognition receptors. Despite its improved tolerability and translation, the use of nucleoside-modified mRNA largely compromises innate immune responses and weakens dendritic cell (DC) activation that are critical to mount a strong adaptive immunity to vaccination. There is thus a need in the art for LNP compositions with enhanced adjuvanticity and methods of use thereof. The present disclosure addresses this need. BRIEF SUMMARY OF THE DISCLOSURE In one aspect, the disclosure provides a compound of Formula (I), or a salt, solvate, stereoisomer, or isotopologue thereof, wherein R¹, R², R^3a, R^3b, R^4a, R^4b, R^4c, and R^4d are defined elsewhere herein: . In certain embodiments, (I) is a compound of Formula (Ia), or a salt, solvate, stereoisomer, or isotopologue thereof, wherein R¹, R², R^3a, R^3b, R^4a, R^4b, R^4c, R^4d, R^6a, R^6b, and L are defined elsewhere herein: . In certain embodiments, (I) is (C12-TLRa): . In - 2 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) . hydrocarbyl substituent; (b) at least one ionizable lipid; (c) at least one helper lipid; (d) cholesterol; and (e) at least one polymer conjugated lipid. In another aspect, the disclosure provides a pharmaceutical composition comprising the lipid nanoparticle (LNP) of the disclosure and a pharmaceutically acceptable carrier. In another aspect, the disclosure provides a method of generating an innate immune response in a subject, the method comprising administering to the subject the lipid nanoparticle (LNP) of the disclosure or the pharmaceutical composition of the disclosure. In another aspect, method of treating, preventing, and/or ameliorating an infection, disease, or disorder in a subject, the method comprising administering to the subject a lipid nanoparticle (LNP) of the disclosure, wherein the LNP comprises at least one cargo molecule. 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. FIG.1 provides a scheme depicting an adjuvant lipidoid-substituted SARS-CoV-2 mRNA-LNP vaccine and its proposed mechanism of action to elicit potent adaptive immunity. In one non-limiting embodiment, an exemplary SARS-CoV-2 mRNA-LNP vaccine is formulated by adjuvant lipidoid and/or additional lipidoid, DOPE, DMG-PEG, and cholesterol, along with m1ψ-modified SARS-CoV-2 mRNA encoding the diproline modified spike glycoprotein. After injection, the vaccine is internalized by dendritic cells (DCs), where adjuvant lipidoid agonizes endosomal-localized TLR7/8 to activate DCs and mRNA is translated into spike antigen that is processed and presented by DCs. After migration into the _{draining lymph node, DCs orchestrate robust adaptive immunity together with CD4} ⁺ _{T cells,} - 3 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) _CD8 ⁺ _{T cells, and B cells, including neutralizing antibody (NAb) responses, Th1-biased CD4} ⁺ _{and CD8} ⁺ _{T cell responses, B cell responses, and long-lived plasma cell (LLPC)} responses. FIGs.2A-2I depict the synthesis and characterization of exemplary lipidoids and LNPs. FIG.2A: synthesis of adjuvant lipidoid C12-TLRa by ring-opening reaction between TLR7/8 agonist 1 and C12 epoxide. FIG.2B: C12-TLRa LNP-mediated mLuc delivery (n = 3). DC2.4 cells were treated with mLuc-loaded C12-TLRa LNP (10-40 ng/well) or free mRNA (40 ng/well) for 24 h. FIG.2C: polyamines and the representative synthesis of polyamine-derived lipidoid C12-113. FIG.2D: comparision of C12-TLRa with polyamine- derived lipidoids for in vitro mLuc delivery (n=3). DC2.4 cells were treated with different mLuc-loaded LNPs (10 ng/well) or free mRNA (10 ng/well) for 24 h. FIG.2E: optimization of C12-113/TLRa LNP formulation (n=3). DC2.4 cells were treated with mLuc-loaded C12- 113/TLRa LNP (10 ng/well) with 1-17.5 mol % of C12-TLRa substitution for 24 h. FIG.2F: TLR7-agonistic activity of LNPs measured in HEK-Blue reporter cells (n = 2). Cells were treated with LNPs at different mRNA concentrations for 24 h. FIG.2G: a representative transmission electron microscopy (TEM) image of C12-113/TLRa LNP. Scale bar, 100 nm. Data are presented as mean ± SD. FIGs.2H-2I: synthesis and characterization of adjuvant lipidoid O12-TLRa. FIGs.3A-3H depict adjuvant lipidoid-substituted LNP-mediated in vitro mRNA delivery and DC activation. FIGs.3A-3B: dose-dependent mLuc delivery; DC2.4 cells (FIG. 3A) and BMDCs (FIG.3B) were treated with mLuc-loaded C12-113 LNP or C12-113/TLRa LNP for 24 h. FIG.3C: representative confocal images of cellular uptake. DC2.4 cells were treated with DiO-labelled LNPs for 2 h before staining with LysoTracker Deep Red and Hoechst 33342. Scale bars, 20 μm. FIG.3D: provides a scheme depicting the proposed mechanism of enhanced endosomal escape by C12-113/TLRa LNP. The agonist-receptor interaction between C12-TLRa and TLR7/8 enhances LNP-mediated endosomal disruption. FIGs.3E-3F: flow cytometry analysis of DC maturation. DC2.4 cells (FIG.3E) and BMDCs (FIG.3F) were treated with SARS-CoV-2 mRNA-loaded LNPs (500 ng/mL) for 24 h and then stained with DC maturation markers CD80 and CD86. FIGs.3G-3H: ELISA analysis of proinflammatory cytokines (TNF-α, IL-12p70 and IL-1β). DC2.4 cells (FIG.3G) and BMDCs (FIG.3H) were treated with SARS-CoV-2 mRNA-loaded LNPs (500 ng/mL) for 24 h before ELISA analysis of cytokine production. Data are presented as mean ± SD (n = 3). FIGs.4A-4H depict adjuvant lipidoid substituted LNP-mediated in vivo mRNA delivery and innate immune responses. FIG.4A: in vivo bioluminescence imaging at 6 and 24 - 4 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) h post-treatment of mLuc-loaded LNPs. Mice were s.c. injected with mLuc-loaded LNPs (5 μg mRNA/mouse) at the tail base. Total flux at the injection site and two iLNs was quantified. The dashed circle indicates iLN. FIG.4B: ex vivo fluorescence and luminescence imaging. Mice were s.c. injected with DiR-labelled, mLuc-loaded LNPs (5 μg mRNA/mouse) at the tail base 24 h before euthanasia. Major organs and iLNs were collected for ex vivo imaging. FIG.4C: quantification of fluorescence and luminescence signals in (FIG.4B). FIGs.4D-4E: flow cytometry analysis of matured DCs in iLNs. At 24 h post-injection of SARS-CoV-2 mRNA-loaded LNPs, two iLNs from each mouse were harvested and processed to generate single cell suspensions that were stained with CD11c, CD80, and CD86. FIGs.4F-4H: ELISA analysis of intranodal proinflammatory cytokine production. Two iLNs from each mouse were harvested at 6 or 24 h post-injection of SARS-CoV-2 mRNA-loaded LNPs and processed to generate a single cell suspension that was cultured for 8 h before the supernatant was collected for ELISA analysis of TNF-α (FIG.4F), IL-12p70 (FIG.4G) and IL-1β (FIG.4H). Data are presented as mean ± SD (n = 3). FIGs.5A-5D depict cellular immune responses induced by a non-limiting exemplary adjuvant lipidoid-substituted SARS-CoV-2 mRNA-LNP vaccine. FIG.5A: scheme of prime and boost vaccination strategy and the analysis of T cell responses. C57BL/6J mice were s.c. immunized twice with 5 μg of C12-113 mRNA-LNP or C12-113/TLRa mRNA-LNP vaccine on Day 0 and 21. FIGs.5B-5C: flow cytometry analysis of RBD-specific CD4⁺ and CD8⁺ T cell responses. On Day 35, spleens were harvested and processed to generate single cell suspensions that were stimulated with SARS-CoV-2 RBD peptide pools. T cells were stained for cytolytic marker CD107α, Th1 (IFN-γ, IL-2 and TNF-α), Th2 (IL-4, IL-5) and Th17 (IL- _{17a) intracellular cytokine expression. FIG. 5D: polyfunctional CD4} ⁺ _{and CD8} ⁺ _{T cells. Data} are presented as mean ± SD (n = 4). FIGs.6A-6H depicts humoral immune responses induced by a non-limiting exemplary adjuvant lipidoid-substituted SARS-CoV-2 mRNA-LNP vaccine. FIG.6A: scheme of prime and boost vaccination strategy and the analysis of humoral immune responses. C57BL/6J mice were s.c. immunized twice with 5 μg of SARS-CoV-2 mRNA- LNP vaccine on Day 0 and 21. FIG.6B: RBD-specific IgG titer (n = 7). Serum was collected from vaccinated mice on Day 35 and RBD-specific IgG levels were determined by endpoint dilution ELISA. FIG.6C: neutralizing antibody (NAb) titer (n = 5). Serum was collected from vaccinated mice on Day 35 and NAb levels were measured by a VSV-based pseudovirus neutralization assay. FIG.6D: number of RBD-specific B cells (n=4). On day 35, spleens were harvested and processed to generate single cell suspensions that were - 5 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) stimulated with SARS-CoV-2 RBD peptide pools. Isotype-switched (IgD-IgM-) RBD- specific B cells were analyzed by flow cytometry. FIG.6E: number of RBD-specific B cells by germinal center (GC) or memory phenotype (n = 4). GC B cells were defined as CD38- _GL7 ⁺ _{. Memory B cells were defined as CD38} ⁺ _GL7- _{and subsetted by PD-L2 and CD80} expression. FIGs.6F-6H: quantification of RBD-specific IgG1, IgG2b, and IgG2c antibody- secreting cells (ASCs) in bone marrow (BM) by ELISPOT (n=3). Data are presented as mean ± SD. FIGs.7A-7F depict interactions between agonists and TLR7. FIG.7A: 2D binding pose for TLR7/8 agonist 1. Multiple interactions were observed between TLR7/8 agonist 1 _{and TLR7 at the 1} ^st _{binding site of TLR7. FIG. 7B: 3D binding pose for TLR7/8 agonist 1.} FIG.7C: front (left) and side (right) views of the TLR7/8 agonist 1-bound TLR7 dimer. TLR7 dimer and its dimerization partner are colored green and cyan, respectively. The C atoms of TLR7/8 agonist 1 are colored black. FIG.7D: 2D binding pose for C12-TLRa. _{Multiple interactions were observed between C12-TLRa and TLR7 at the 1st binding site of} TLR7, which were very similar to (FIG.7A). FIG.7E: 3D binding pose for C12-TLRa. FIG. 7F: front (left) and side (right) views of the C12-TLRa-bound TLR7 dimer. TLR7 dimer and its dimerization partner are colored green and cyan, respectively. The C atoms of C12- TLRa are colored black. FIG.8: TLR7-agonistic activity of C12-TLRa evaluated by the HEK-mTLR7 cell assay. The upper image is a photograph of a 96-well plate. Data are presented as mean ± SD (n = 3). FIG.9: TNF-α production in LNP-treated DC2.4 cells.10,000 cells in each well were treated with mRNA-loaded LNPs for 24 h. The supernatant was harvested for ELISA analysis. Data are presented as mean ± SD (n = 3). FIG.10: TLR7-agonistic activity of MC3 LNP and MC3/TLRa LNP measured in HEK-Blue reporter cells. Cells were treated with LNPs at different mRNA concentrations for 24 h. Data are presented as mean ± SD (n = 3). FIG.11: representative TEM image of C12-113 LNP. Scale bar, 100 nm. FIGs.12A-12B depict cell viability. FIG.12A: cell viability of DC2.4 cells at 24 h post-treatment with mLuc- loaded LNPs. FIG.12B: cell viability of BMDCs at 24 h post- treatment with mLuc-loaded LNPs. Data are presented as mean ± SD (n = 3). FIG.13 depicts dose-dependent mLuc delivery with non-limiting exemplary LNPs of the present disclosure. DC2.4 cells were treated with mLuc-loaded MC3 LNP or MC3/TLRa - 6 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) LNP for 24 h. Data are presented as mean ± SD (n = 3). FIGs.14A-14B depict gating strategies for matured DCs. FIG.14A: gating strategy _{for CD80} ⁺ _CD86 ⁺ _{DC2.4 cells. DC2.4 cells were treated with SARS-CoV-2 mRNA-loaded} LNPs (500 ng/mL) for 24 h and then stained with DC maturation markers CD80 and CD86. _{FIG. 14B: gating strategy for CD80} ⁺ _CD86 ⁺ _{BMDCs. BMDCs were treated with SARS-CoV-} 2 mRNA-loaded LNPs (500 ng/mL) for 24 h and then stained with DC surface maker CD11c and DC maturation markers CD80 and CD86. FIG.15 depicts in vivo and ex vivo luminescence imaging at 24 h post-injection of non-limiting exemplary LNPs of the present disclosure. Mice were s.c. injected with mLuc- loaded LNPs (5 μg mRNA/mouse) at the tail base. The interval of different batches was over one month. The LNPs from Batch 1 were used in FIGs.4A-4B. C12-113/TLRa LNP consistently outperformed C12-113 LNP in transfecting injection sites and iLNs in different LNP batches. FIGs.16A-16B depict in vivo expression kinetics of mLuc-LNPs after s.c. injection at tail bases. FIG.16A: consecutive in vivo luminescence imaging of mice treated with C12-113 LNP (left) or C12- 113/TLRa LNP (right) at an dose of 5 μg mRNA per mouse. FIG.16B: quantification of total flux at tail bases. Data are presented as mean ± SD (n = 3). FIG.17 depicts gating strategies for intralymphatic matured DCs. At 24 h post- injection of SARS-CoV-2 mRNA-loaded LNPs (5 μg mRNA/mouse), two iLNs from each mouse were harvested and processed to generate single cell suspensions that were stained with DC surface maker CD11c and DC maturation markers CD80 and CD86. CD80⁺CD11c⁺ and CD86⁺CD11c⁺ cells were gated. FIGs.18A-18C provides graphs depicting serum proinflammatory cytokine levels of TNF-α (FIG.18A), IL12p70 (FIG.18B), and IL-1β (FIG.18C). Serum from immunized mice were harvested at 6 or 24 h post-injection of SARS-CoV-2 mRNA-loaded LNPs (5 μg mRNA/mouse). Data are presented as mean ± SD (n = 3). FIG.19 provides representative photographs of immunized mice. C57BL/6J mice were s.c. immunized twice with 5 μg of C12-113 mRNA-LNP or C12-113/TLRa mRNA- LNP vaccine on Day 0 and 21. No skin appearance abnormality (e.g., rash and damage) at tail bases was observed after prime and boost vaccination for both LNPs. FIG.20 depicts weight changes observed in exemplary mouse studies. C57BL/6J mice were s.c. immunized twice with 5 μg of C12-113 mRNA-LNP or C12-113/TLRa mRNA-LNP vaccine on Day 0 and 21. Arrows indicate the days of immunization. No body weight loss was observed for all groups. Data are presented as mean ± SD (n = 4). - 7 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) FIGs.21A-21C: gating strategy for splenic T cells. FIG.21A: gating scheme for the _{identification of CD4} ⁺ _{and CD8} ⁺ _{T cells. FIG. 21B: identification of antigen specific} cytokine-expressing CD4⁺ T cells. FIG.21C: identification of antigen specific cytokine- expressing CD8⁺ T cells. FIG.22 depicts subsets of RBD-specific IgG and IgG2c/IgG1 ratio. C57BL/6J mice were s.c. immunized twice with 5 μg of SARS-CoV-2 mRNA-LNP vaccine on Day 0 and 21. Serum was collected from vaccinated mice on Day 35 and RBD-specific IgG1 and IgG2 were analyzed by ELISA (n = 2-4). FIGs.23A-23C: ating strategy for splenic B cells. (A) Gating scheme for the _{identification of isotype-switched (IgD}-_IgM-_{) splenic B cells. (B) Identification of IgD}-_IgM- RBD-binding B cells using biotinylated protein probes. (C) Identification of germinal center _(CD38-_GL7 ⁺ _{) and memory phenotype (CD38} ⁺ _GL7-_{) subsets in total IgD}-_IgM- _{and IgD}- _IgM- _RBD ⁺ _{B cells. Cells with memory phenotype are further subsetted by the memory markers} PD-L2 and CD80 expression. DETAILED DESCRIPTION OF THE DISCLOSURE 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 - 8 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) least one of A or B" has the same meaning as "A, B, or A and B." In addition, it is to be understood that the phraseology or terminology employed herein, and not otherwise defined, is for the purpose of description only and not of limitation. Any use of section headings is intended to aid reading of the document and is not to be interpreted as limiting; information that is relevant to a section heading may occur within or outside of that particular section. All publications, patents, and patent documents referred to in this document are incorporated by reference herein in their entirety, as though individually incorporated by reference. In the methods described herein, the acts can be carried out in any order, except when a temporal or operational sequence is explicitly recited. Furthermore, specified acts can be carried out concurrently unless explicit claim language recites that they be carried out separately. For example, a claimed act of doing X and a claimed act of doing Y can be conducted simultaneously within a single operation, and the resulting process will fall within the literal scope of the claimed process. Description Lipid nanoparticle (LNP)-formulated mRNA vaccines represent a promising tool to prevent infectious diseases as exemplified by the recent success of SARS-CoV-2 mRNA vaccines. To avoid the immune-stimulatory effect and uncontrollable inflammation, immune silent nucleoside-modified mRNAs are preferentially used. However, such modification largely abrogates the innate immune responses that are critical to orchestrating a robust adaptive immunity. In one aspect, the present disclosure describes the development of a novel LNP component (i.e., adjuvant lipidoid) which can enhance the adjuvanticity of LNP vaccines (e.g., SARS-CoV-2 mRNA LNP vaccines), and LNPs comprising the same. The present disclosure demonstrates that partial substitution of ionizable lipidoid with adjuvant lipidoid in LNP formulations not only enhanced mRNA delivery, but also endowed LNPs with Toll-like receptor 7/8-agonistic activity, which significantly increased the innate immunity of mRNA-LNP vaccine without causing noticeable adverse effects. Moreover, this vaccine elicited broadly neutralizing antibodies against multiple SARS-CoV-2 pseudovirus variants, strong Th1-biased cellular immunity, as well as robust B cell and long-lived plasma cell responses. Overall, the present disclosure describes a novel and safe LNP formulation paradigm to enhance the immunogenicity of mRNA vaccines. Definitions - 9 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) The term "about" as used herein can allow for a degree of variability in a value or range, for example, within 10%, within 5%, or within 1% of a stated value or of a stated limit of a range, and includes the exact stated value or range. The term "alkenyl" as used herein refers to straight and branched chain and cyclic alkyl groups as defined herein, except that at least one double bond exists between two carbon atoms. Thus, alkenyl groups have from 2 to 40 carbon atoms, or 2 to about 20 carbon atoms, or 2 to 12 carbon atoms or, in some embodiments, from 2 to 8 carbon atoms. Examples include, but are not limited to vinyl, -CH=C=CCH₂, -CH=CH(CH₃), - CH=C(CH3)2, -C(CH3)=CH2, -C(CH3)=CH(CH3), -C(CH2CH3)=CH2, cyclohexenyl, cyclopentenyl, cyclohexadienyl, butadienyl, pentadienyl, and hexadienyl among others. The term "alkoxy" as used herein refers to an oxygen atom connected to an alkyl group, including a cycloalkyl group, as are defined herein. Examples of linear alkoxy groups include but are not limited to methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, and the like. Examples of branched alkoxy include but are not limited to isopropoxy, sec-butoxy, tert-butoxy, isopentyloxy, isohexyloxy, and the like. Examples of cyclic alkoxy include but are not limited to cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and the like. An alkoxy group can include about 1 to about 12, about 1 to about 20, or about 1 to about 40 carbon atoms bonded to the oxygen atom, and can further include double or triple bonds, and can also include heteroatoms. For example, an allyloxy group or a methoxyethoxy group is also an alkoxy group within the meaning herein, as is a methylenedioxy group in a context where two adjacent atoms of a structure are substituted therewith. The term "alkyl" as used herein refers to straight chain and branched alkyl groups and cycloalkyl groups having from 1 to 40 carbon atoms, 1 to about 20 carbon atoms, 1 to 12 carbons or, in some embodiments, from 1 to 8 carbon atoms. Examples of straight chain alkyl groups include those with from 1 to 8 carbon atoms such as methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl groups. Examples of branched alkyl groups include, but are not limited to, isopropyl, iso-butyl, sec-butyl, t-butyl, neopentyl, isopentyl, and 2,2- dimethylpropyl groups. As used herein, the term "alkyl" encompasses n-alkyl, isoalkyl, and anteisoalkyl groups as well as other branched chain forms of alkyl. Representative substituted alkyl groups can be substituted one or more times with any of the groups listed herein, for example, amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups. The term "alkynyl" as used herein refers to straight and branched chain alkyl groups, except that at least one triple bond exists between two carbon atoms. Thus, alkynyl groups have from 2 to 40 carbon atoms, 2 to about 20 carbon atoms, or from 2 to 12 carbons or, in - 10 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) 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 disclosure includes, but is not limited to, the use of partial nucleotide sequences of more than one gene and that these nucleotide sequences are arranged in various combinations to elicit the desired immune response. Moreover, a skilled artisan will understand that an antigen need not be encoded by a "gene" at all. It is readily apparent that an antigen can be generated synthesized or can be derived from a biological sample. Such a biological sample can include, but is not limited to a tissue sample, a tumor sample, a cell or a biological fluid. The term "amine" as used herein refers to primary, secondary, and tertiary amines having, e.g., the formula N(group)3 wherein each group can independently be H or non-H, such as alkyl, aryl, and the like. Amines include but are not limited to R-NH₂, for example, - 11 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) 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 "aryl-(C1-C6)alkyl" or “aralkyl” as used herein refers to a functional group wherein a one to six carbon alkylene chain is attached to an aryl group, e.g., -CH₂CH₂-phenyl or -CH2-phenyl (or benzyl). Specific examples are aryl-CH2- and aryl-CH(CH3)-. The term "substituted aryl-(C₁-C₆)alkyl" refers to an aryl-(C₁-C₆)alkyl functional group in which the aryl group is substituted. A specific example is substituted aryl(CH2)-. Similarly, the term "heteroaryl-(C₁-C₆)alkyl" refers to a functional group wherein a one to three carbon alkylene chain is attached to a heteroaryl group, e.g., -CH2CH2-pyridyl. A specific example is - 12 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) heteroaryl-(CH2)-. The term "substituted heteroaryl-(C1-C6)alkyl" or "heteroaralkyl" refers to a heteroaryl-(C₁-C₆)alkyl functional group in which the heteroaryl group is substituted. A specific example is substituted heteroaryl-(CH2)-. The term "cationic lipid" refers to any of a number of lipid species that carry a net positive charge at a selected pH, such as physiological pH (e.g., pH of about 7.0). It has been found that cationic lipids comprising alkyl chains with multiple sites of unsaturation, e.g., at least two or three sites of unsaturation, are particularly useful for forming lipid particles with increased membrane fluidity. A number of cationic lipids and related analogs, which are also useful in the present disclosure, have been described in U.S. Patent Publication Nos. 20060083780 and 20060240554; U.S. Pat. Nos.5,208,036; 5,264,618; 5,279,833; 5,283,185; 5,753,613; and 5,785,992; and PCT Publication Nos. WO1996/010390, WO2021077066, WO2021077067, WO2023015200, WO2023056282, and WO2023056418, 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 cationic 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 "conjugated lipid" as used herein refers to a lipid which is conjugated to one or more polymeric groups, which inhibits aggregation of lipid particles. Such lipid conjugates include, but are not limited to, polyamide oligomers (e.g., ATTA-lipid conjugates), PEG-lipid conjugates, such as PEG coupled to dialkyloxypropyls, PEG coupled to diacylglycerols, PEG coupled to cholesterol, PEG coupled to phosphatidylethanolamines, PEG conjugated to ceramides (e.g., U.S. Pat. No.5,885,613, the disclosure of which is herein incorporated by reference in its entirety for all purposes), cationic PEG lipids, and mixtures thereof. PEG can be conjugated directly to the lipid or may be linked to the lipid via a linker moiety. Any linker moiety suitable for coupling the PEG to a lipid can be used including, e.g., non-ester containing linker moieties and ester-containing linker moieties. In preferred embodiments, non-ester containing linker moieties are used. 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 - 13 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) such as, but not limited to, decalinyl, and the like. Cycloalkyl groups also include rings that are substituted with straight or branched chain alkyl groups as defined herein. Representative substituted cycloalkyl groups can be mono-substituted or substituted more than once, such as, but not limited to, 2,2-, 2,3-, 2,4- 2,5- or 2,6-disubstituted cyclohexyl groups or mono-, di- or tri-substituted norbornyl or cycloheptyl groups, which can be substituted with, for example, amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups. The term "cycloalkenyl" alone or in combination denotes a cyclic alkenyl group. A "disease" is a state of health of an animal wherein the animal cannot maintain homeostasis, and wherein if the disease is not ameliorated then the animal's health continues to deteriorate. In contrast, a "disorder" in an animal is a state of health in which the animal is able to maintain homeostasis, but in which the animal's state of health is less favorable than it would be in the absence of the disorder. Left untreated, a disorder does not necessarily cause a further decrease in the animal's state of health. A disease or disorder is "alleviated" if the severity of a symptom of the disease or disorder, the frequency with which such a symptom is experienced by a patient, or both, is reduced. As used herein, the terms "effective amount," "pharmaceutically effective amount" and "therapeutically effective amount" refer to a nontoxic but sufficient amount of an agent to provide the desired biological result. That result may be reduction and/or alleviation of the signs, symptoms, or causes of a disease, or any other desired alteration of a biological system. An appropriate therapeutic amount in any individual case may be determined by one of ordinary skill in the art using routine experimentation. In particular, in the case of a mRNA, and "effective amount" or "therapeutically effective amount" of a therapeutic nucleic acid as relating to a mRNA is an amount sufficient to produce the desired effect, e.g., mRNA-directed expression of an amount of a protein that causes a desirable biological effect in the organism within which the protein is expressed. For 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 - 14 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) produces a similar level of expression as observed in a healthy individual in an individual with aberrant expression of the protein (i.e., protein deficient individual). Suitable assays for measuring the expression of an mRNA or protein include, but are not limited to dot blots, Northern blots, in situ hybridization, ELISA, immunoprecipitation, enzyme function, as well as phenotypic assays known to those of skill in the art. The term "encode" as used herein refers to the product specified (e.g., protein and RNA) by a given sequence of nucleotides in a nucleic acid (i.e., DNA and/or RNA), upon transcription or translation of the DNA or RNA, respectively. In certain embodiments, the term "encode" refers to the RNA sequence specified by transcription of a DNA sequence. In certain embodiments, the term "encode" refers to the amino acid sequence (e.g., polypeptide or protein) specified by translation of mRNA. In certain embodiments, the term "encode" refers to the amino acid sequence specified by transcription of DNA to mRNA and subsequent translation of the mRNA encoded by the DNA sequence. In certain embodiments, the encoded product may comprise a direct transcription or translation product. In certain embodiments, the encoded product may comprise post-translational modifications understood or reasonably expected by one skilled in the art. The term "fully encapsulated" indicates that the active agent or therapeutic agent in the lipid particle is not significantly degraded after exposure to serum or a nuclease or protease assay that would significantly degrade free DNA, RNA, or protein. In a fully encapsulated system, preferably less than about 25% of the active agent or therapeutic agent in the particle is degraded in a treatment that would normally degrade 100% of free active agent or therapeutic agent, more preferably less than about 10%, and most preferably less than about 5% of the active agent or therapeutic agent in the particle is degraded. In the context of nucleic acid therapeutic agents, full encapsulation may be determined by an OLIGREEN® assay. OLIGREEN® is an ultra-sensitive fluorescent nucleic acid stain for quantitating oligonucleotides and single-stranded DNA or RNA in solution (available from Invitrogen Corporation; Carlsbad, Calif.). "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 - 15 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) groups, wherein all hydrogen atoms are replaced by halogen atoms, such as fluoro. Examples of haloalkyl include trifluoromethyl, 1,1-dichloroethyl, 1,2-dichloroethyl, 1,3-dibromo-3,3- difluoropropyl, perfluorobutyl, and the like. The term "helper lipid" as used herein refers to a lipid capable of increasing the effectiveness of delivery of lipid-based particles such as cationic lipid-based particles to a target, preferably into a cell. The helper lipid can be neutral, positively charged, or negatively charged. In certain embodiments, the helper lipid is neutral or negatively charged. Non- limiting examples of helper lipids include 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphoethanolamine (DOPE), 1-palmitoyl- 2-oleoyl-sn-glycero-3phosphocholin (POPC) and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC). The term "heteroalkyl" as used herein by itself or in combination with another term, means, unless otherwise stated, a non-cyclic stable straight or branched chain, or combinations thereof, including at least one carbon atom and at least one heteroatom selected from the group consisting of O, N, P, Si, and S, and wherein the nitrogen and sulfur atoms may optionally be oxidized, and the nitrogen heteroatom may optionally be quaternized. The heteroatom(s) (e.g., O, N, P, and S) may be placed at any interior position of the heteroalkyl group or at either terminal position at which the group is attached to the remainder of the molecule. The term "heteroaryl" as used herein refers to aromatic ring compounds containing 5 or more ring members, of which, one or more is a heteroatom such as, but not limited to, N, O, and S; for instance, heteroaryl rings can have 5 to about 8-12 ring members. A heteroaryl group is a variety of a heterocyclyl group that possesses an aromatic electronic structure. A heteroaryl group designated as a C2-heteroaryl can be a 5-ring with two carbon atoms and three heteroatoms, a 6-ring with two carbon atoms and four heteroatoms and so forth. Likewise a C4-heteroaryl can be a 5-ring with one heteroatom, a 6-ring with two heteroatoms, 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 - 16 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) substituted heteroaryl groups can be substituted one or more times with groups such as those listed herein. Additional examples of aryl and heteroaryl groups include but are not limited to phenyl, biphenyl, indenyl, naphthyl (1-naphthyl, 2-naphthyl), N-hydroxytetrazolyl, N- hydroxytriazolyl, N-hydroxyimidazolyl, anthracenyl (1-anthracenyl, 2-anthracenyl, 3- anthracenyl), thiophenyl (2-thienyl, 3-thienyl), furyl (2-furyl, 3-furyl) , indolyl, oxadiazolyl, isoxazolyl, quinazolinyl, fluorenyl, xanthenyl, isoindanyl, benzhydryl, acridinyl, thiazolyl, pyrrolyl (2-pyrrolyl), pyrazolyl (3-pyrazolyl), imidazolyl (1-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl), triazolyl (1,2,3-triazol-1-yl, 1,2,3-triazol-2-yl 1,2,3-triazol-4-yl, 1,2,4-triazol-3-yl), oxazolyl (2-oxazolyl, 4-oxazolyl, 5-oxazolyl), thiazolyl (2-thiazolyl, 4- thiazolyl, 5-thiazolyl), pyridyl (2-pyridyl, 3-pyridyl, 4-pyridyl), pyrimidinyl (2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, 6-pyrimidinyl), pyrazinyl, pyridazinyl (3- pyridazinyl, 4- pyridazinyl, 5-pyridazinyl), quinolyl (2-quinolyl, 3-quinolyl, 4-quinolyl, 5-quinolyl, 6- quinolyl, 7-quinolyl, 8-quinolyl), isoquinolyl (1-isoquinolyl, 3-isoquinolyl, 4-isoquinolyl, 5- isoquinolyl, 6-isoquinolyl, 7-isoquinolyl, 8-isoquinolyl), benzo[b]furanyl (2-benzo[b]furanyl, 3-benzo[b]furanyl, 4-benzo[b]furanyl, 5-benzo[b]furanyl, 6-benzo[b]furanyl, 7- benzo[b]furanyl), 2,3-dihydro-benzo[b]furanyl (2-(2,3-dihydro-benzo[b]furanyl), 3-(2,3- dihydro-benzo[b]furanyl), 4-(2,3-dihydro-benzo[b]furanyl), 5-(2,3-dihydro-benzo[b]furanyl), 6-(2,3-dihydro-benzo[b]furanyl), 7-(2,3-dihydro-benzo[b]furanyl), benzo[b]thiophenyl (2- benzo[b]thiophenyl, 3-benzo[b]thiophenyl, 4-benzo[b]thiophenyl, 5-benzo[b]thiophenyl, 6- benzo[b]thiophenyl, 7-benzo[b]thiophenyl), 2,3-dihydro-benzo[b]thiophenyl, (2-(2,3- dihydro-benzo[b]thiophenyl), 3-(2,3-dihydro-benzo[b]thiophenyl), 4-(2,3-dihydro- benzo[b]thiophenyl), 5-(2,3-dihydro-benzo[b]thiophenyl), 6-(2,3-dihydro- benzo[b]thiophenyl), 7-(2,3-dihydro-benzo[b]thiophenyl), indolyl (1-indolyl, 2-indolyl, 3-indolyl, 4-indolyl, 5-indolyl, 6-indolyl, 7-indolyl), indazole (1-indazolyl, 3-indazolyl, 4-indazolyl, 5-indazolyl, 6-indazolyl, 7-indazolyl), benzimidazolyl (1-benzimidazolyl, 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, - 17 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) 10,11-dihydro-5H-dibenz[b,f]azepine-4-yl, 10,11-dihydro-5H-dibenz[b,f]azepine-5-yl), and the like. The term "heterocycloalkyl" as used herein refers to an aliphatic, partially unsaturated or fully saturated, 3- to 14-membered ring system, including single rings of 3 to 8 atoms and bi- and tricyclic ring systems where at least one of the carbon atoms of the ring is replaced with a heteroatom such as, but not limited to, nitrogen, oxygen, sulfur, or phosphorus. A heterocycloalkyl can include one to four heteroatoms independently selected from oxygen, nitrogen, and sulfur, wherein a nitrogen and sulfur heteroatom optionally can be oxidized and a nitrogen heteroatom optionally can be substituted. Representative heterocycloalkyl groups include, but are not limited, to the following exemplary groups: pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, and tetrahydrofuryl. The term "heterocyclyl" as used herein refers to aromatic and non-aromatic ring compounds containing three or more ring members, of which one or more is a heteroatom such as, but not limited to, N, O, and S. Thus, a heterocyclyl can be a cycloheteroalkyl, or a heteroaryl, or if polycyclic, any combination thereof. In some embodiments, heterocyclyl groups include 3 to about 20 ring members, whereas other such groups have 3 to about 15 ring members. A heterocyclyl group designated as a C2-heterocyclyl can be a 5-ring with two carbon atoms and three heteroatoms, a 6-ring with two carbon atoms and four heteroatoms and so forth. Likewise a C4-heterocyclyl can be a 5-ring with one heteroatom, a 6-ring with two heteroatoms, and so forth. The number of carbon atoms plus the number of heteroatoms equals the total number of ring atoms. A heterocyclyl ring can also include one or more double bonds. A heteroaryl ring is an embodiment of a heterocyclyl group. The phrase "heterocyclyl group" includes fused ring species including those that include fused aromatic and non-aromatic groups. For example, a dioxolanyl ring and a benzdioxolanyl ring system (methylenedioxyphenyl ring system) are both heterocyclyl groups within the meaning herein. 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, - 18 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) 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- C_b)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-C_b)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, 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 X³ are independently selected from noble gases" would include the scenario where, for example, X¹, X², and X³ are all the same, where X¹, X², and X³ are all different, where X¹ and X² are the same but X³ is different, and other analogous permutations. The term "ionizable lipid" as used herein refers to a lipid (e.g., a cationic lipid) 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 pK_a 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 - 19 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) 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. In the present context, the term "lipidoid" refers to any compound having the characteristics of a lipid. In certain embodiments, lipidoids may have a series of secondary and tertiary amines, which increases the net positive charge of the compound. 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 - 20 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) are known to those of skill in the art. For example, various in vitro transfer kits are commercially available. According to the present disclosure, mRNA can be modified by further stabilizing modifications and cap formation in addition to the modifications according to the disclosure. 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. 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 - 21 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) 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" 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. The terms "spacer" or "spacer element" as used herein with reference to a crRNA or - 22 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) sgRNA, refers to the polynucleotide sequence that can specifically hybridize to a target nucleic acid sequence. The spacer element interacts with the target nucleic acid sequence through hydrogen bonding between complementary base pairs (i.e., paired bases). A spacer element binds to a selected DNA target sequence. Accordingly, the spacer element is a DNA target-binding sequence. The spacer element determines the location of Cas protein's site- specific binding and endonucleolytic cleavage. Spacer elements range from -17- to -84 nucleotides in length, depending on the Cas protein with which they are associated, and have an average length of 36 nucleotides. For example, for SpyCas9, the functional length for a spacer to direct specific cleavage is typically about 12-25 nucleotides. Variability of the functional length for a spacer element is known in the art, as indicated in U.S. Published Patent Application No.2014/0315985, which is incorporated herein by reference in its entirety. Suitable pharmaceutically acceptable acid addition salts may be prepared from an inorganic acid or from an organic acid. Examples of inorganic acids include hydrochloric, hydrobromic, hydriodic, nitric, carbonic, sulfuric (including sulfate and hydrogen sulfate), and phosphoric acids (including hydrogen phosphate and dihydrogen phosphate). Appropriate organic acids may be selected from aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic and sulfonic classes of organic acids, examples of which include formic, acetic, propionic, succinic, glycolic, gluconic, lactic, malic, tartaric, citric, ascorbic, glucuronic, maleic, malonic, saccharin, fumaric, pyruvic, aspartic, glutamic, benzoic, anthranilic, 4-hydroxybenzoic, phenylacetic, mandelic, embonic (pamoic), methanesulfonic, ethanesulfonic, benzenesulfonic, pantothenic, trifluoromethanesulfonic, 2- hydroxyethanesulfonic, p-toluenesulfonic, sulfanilic, cyclohexylaminosulfonic, stearic, alginic, β-hydroxybutyric, salicylic, galactaric and galacturonic acid. Suitable pharmaceutically acceptable base addition salts of compounds described herein include, for example, ammonium salts, metallic salts including alkali metal, alkaline earth metal and transition metal salts such as, for example, calcium, magnesium, potassium, sodium and zinc salts. Pharmaceutically acceptable base addition salts also include organic salts made from basic amines such as, for example, N,N'-dibenzylethylene-diamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine) and procaine. All of these salts may be prepared from the corresponding compound by reacting, for example, the appropriate acid or base with the compound. As used herein, the term "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" means a pharmaceutically acceptable material, composition or carrier, - 23 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) 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 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, - 24 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) 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 "placenta" as used herein refers to the maternal organ that connects the developing fetus to the uterine wall. After birth, the placenta is expelled and is referred to as a postpartum placenta. 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 "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 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. - 25 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) The term "substantially" as used herein refers to a majority of, or mostly, as in at least about 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, 99.99%, or at least about 99.999% or more, or 100%. The term "substantially free of" as used herein can mean having none or having a trivial amount of, such that the amount of material present does not affect the material properties of the composition including the material, such that the composition is about 0 wt% to about 5 wt% of the material, or about 0 wt% to about 1 wt%, or about 5 wt% or less, or less than, equal to, or greater than about 4.5 wt%, 4, 3.5, 3, 2.5, 2, 1.5, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.01, or about 0.001 wt% or less. The term "substantially free of" can mean having a trivial amount of, such that a composition is about 0 wt% to about 5 wt% of the material, or about 0 wt% to about 1 wt%, or about 5 wt% or less, or less than, equal to, or greater than about 4.5 wt%, 4, 3.5, 3, 2.5, 2, 1.5, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.01, or about 0.001 wt% or less, or about 0 wt%. The term "substituted" as used herein in conjunction with a molecule or an organic group as defined herein refers to the state in which one or more hydrogen atoms contained therein are replaced by one or more non-hydrogen atoms. The term "functional group" or "substituent" as used herein refers to a group that can be or is substituted onto a molecule or onto an organic group. Examples of substituents or functional groups include, but are not limited to, a halogen (e.g., F, Cl, Br, and I); an oxygen atom in groups such as hydroxy groups, alkoxy groups, aryloxy groups, aralkyloxy groups, oxo(carbonyl) groups, carboxyl groups including carboxylic acids, carboxylates, and carboxylate esters; a sulfur atom in groups such as thiol groups, alkyl and aryl sulfide groups, sulfoxide groups, sulfone groups, sulfonyl groups, and sulfonamide groups; a nitrogen atom in groups such as amines, hydroxyamines, nitriles, nitro groups, N-oxides, hydrazides, azides, and enamines; and other heteroatoms in various other groups. Non-limiting examples of substituents that can be bonded to a substituted carbon (or other) atom include F, Cl, Br, I, OR, OC(O)N(R)₂, CN, NO, NO2, ONO2, azido, CF3, OCF3, R, O (oxo), S (thiono), C(O), S(O), methylenedioxy, ethylenedioxy, N(R)₂, SR, SOR, SO₂R, SO₂N(R)₂, SO₃R, C(O)R, C(O)C(O)R, C(O)CH2C(O)R, C(S)R, C(O)OR, OC(O)R, C(O)N(R)2, OC(O)N(R)2, C(S)N(R)2, (CH2)0- ₂N(R)C(O)R, (CH₂)_0-2N(R)N(R)₂, N(R)N(R)C(O)R, N(R)N(R)C(O)OR, N(R)N(R)CON(R)₂, N(R)SO2R, N(R)SO2N(R)2, N(R)C(O)OR, N(R)C(O)R, N(R)C(S)R, N(R)C(O)N(R)2, N(R)C(S)N(R)₂, N(COR)COR, N(OR)R, C(=NH)N(R)₂, C(O)N(OR)R, and C(=NOR)R, wherein R can be hydrogen or a carbon-based moiety; for example, R can be hydrogen, (C1- C₁₀₀) hydrocarbyl, alkyl, acyl, cycloalkyl, aryl, aralkyl, heterocyclyl, heteroaryl, or heteroarylalkyl; or wherein two R groups bonded to a nitrogen atom or to adjacent nitrogen - 26 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) 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 certain 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 term "toll-like receptor agonist” as used herein refers to a small molecule which agonizes, stimulates, and/or activates a toll-like receptor (TLR). In certain embodiments, agonism, stimulation, and/or activation of one or more TLRs (e.g., TLR7/8) may generate and/or promote an innate immune response. Small molecule TLR agonists are known in the art, non-limiting exemplary TLR agonists including TLR7/8 agonist 1, imiquimod, resiquimod, and gardiquimod. 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. Lipid and Lipidoid Compounds Toll-Like Receptor (TLR) Agonist Lipidoids In one aspect, the present disclosure provides a compound of Formula (I), or a salt, solvate, stereoisomer, or isotopologue thereof: - 27 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) , wherein: R¹ is selected from the group substituted C₁-C₆ alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C₆-C₁₀ aryl, and optionally substituted heteroaryl; R² is selected from the group consisting of H, R⁵, optionally substituted C1-C6 alkyl, optionally substituted C₃-C₈ cycloalkyl, optionally substituted C₂-C₈ heterocycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted heteroaryl; R^3a and R^3b are each independently selected from the group consisting of H, R⁵, R^6a, and optionally substituted C1-C6 alkyl; R^4a, R^4b, R^4c, and R^4d are each independently selected from the group consisting of H, R⁵, halogen, CN, NO2, optionally substituted C1-C6 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C₂-C₈ heterocycloalkyl, optionally substituted C₆-C₁₀ aryl, optionally substituted C2-C10 heteroaryl, OR^A, N(R^A)(R^B), C(=O)N(R^A)(R^B), C(=O)R^A, C(=O)OR^A, OC(=O)R^A, OC(=O)OR^A, SR^A, S(=O)R^A, S(=O)₂R^A, N(R^A)S(=O)₂R^B, wherein at least one of R¹, R², R^3a, R^3b, R^4a, R^4b, R^4c, and R^4d is R⁵, or at least one of R^3a and R^3b is R^6a; each occurrence of R^6a and R^6b is independently selected from the group consisting of - (optionally substituted C₁-C₆ alkylenyl)-C(=O)OR^7a, -(optionally substituted C₁-C₆ alkylenyl)-C(=O)N(R^7a)(R^7b), -(optionally substituted C1-C6 alkylenyl)-C(=O)R^7a, - (optionally substituted C₁-C₆ alkylenyl)-(R^7a), -C(=O)OR^7a, -C(=O)N(R^7a)(R^7b), -C(=O)R^7a, and R^7a; each occurrence of R^7a and R^7b is independently selected from the group consisting of optionally substituted C6-C28 alkyl, optionally substituted C6-C28 heteroalkyl, optionally substituted C₃-C₈ cycloalkyl, optionally substituted C₆-C₈ heterocycloalkyl, optionally substituted C6-C28 alkenyl, and optionally substituted C6-C28 alkynyl; - 28 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) L is selected from the group consisting of a bond, optionally substituted C1-C6 alkylenyl, optionally substituted C₂-C₆ alkenylenyl, optionally substituted C₂-C₆ heteroalkylenyl, optionally substituted C3-C8 cycloalkylenyl, optionally substituted C2-C8 heterocycloalkylenyl, optionally substituted C₇-C₁₂ aralkylenyl, optionally substituted C₅-C₁₂ heteroaralkylenyl, optionally substituted C6-C10 arylenyl, and optionally substituted C2-C10 heteroarylenyl; and each occurrence of R^A and R^B is independently selected from the group consisting of H, C(=O)(optionally substituted C₁-C₆ alkyl), C(=O)(optionally substituted C₃-C₈ cycloalkyl), C(=O)(optionally substituted C2-C8 heterocycloalkyl), C(=O)(optionally substituted C6-C10 aryl), C(=O)(optionally substituted C₂-C₁₀ heteroaryl), optionally substituted C₁-C₆ alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted C₂-C₁₀ heteroaryl. In certain embodiments, each occurrence of optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted alkylenyl, optionally substituted heteroalkylenyl, optionally substituted cycloalkylenyl, optionally substituted heterocycloalkylenyl, optionally substituted aralkylenyl, heteroaralkylenyl, optionally substituted arylenyl, and optionally substituted heteroarylenyl, if present, 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. In certain embodiments, exactly one of R¹, R², R^3a, R^3b, R^4a, R^4b, R^4c, and R^4d is R⁵. In certain embodiments, R¹ is R⁵. In certain embodiments, R¹ is CH₂CH(CH₃)₂. In certain embodiments, R¹ is CH2C(OH)(CH3)2. In certain embodiments, the compound of Formula (I) is a compound of Formula (Ia), or a salt, solvate, stereoisomer, or isotopologue thereof: - 29 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) R² is selected from the C₁-C₆ alkyl, optionally substituted C 3-C8 C2-C8 heterocycloalkyl, optionally substituted C₆-C₁₀ aryl, and optionally substituted heteroaryl; R^3a and R^3b are each independently selected from the group consisting of H, R^6a, and optionally substituted C₁-C₆ alkyl; R^4a, R^4b, R^4c, and R^4d are each independently selected from the group consisting of H, halogen, CN, NO₂, optionally substituted C₁-C₆ alkyl, optionally substituted C₃-C₈ cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C6-C10 aryl, optionally substituted C₂-C₁₀ heteroaryl, OR^A, N(R^A)(R^B), C(=O)N(R^A)(R^B), C(=O)R^A, C(=O)OR^A, OC(=O)R^A, OC(=O)OR^A, SR^A, S(=O)R^A, S(=O)2R^A, N(R^A)S(=O)2R^B, N(R^A)C(=O)R^B, and S(=O)₂N(R^A)(R^B); each occurrence of R^6a and R^6b is independently selected from the group consisting of - (optionally substituted C₁-C₆ alkylenyl)-C(=O)OR^7a, -(optionally substituted C₁-C₆ alkylenyl)-C(=O)N(R^7a)(R^7b), -(optionally substituted C1-C6 alkylenyl)-C(=O)R^7a, - (optionally substituted C₁-C₆ alkylenyl)-(R^7a), -C(=O)OR^7a, -C(=O)N(R^7a)(R^7b), -C(=O)R^7a, and R^7a; each occurrence of R^7a and R^7b is independently selected from the group consisting of optionally substituted C₁-C₂₈ alkyl, optionally substituted C₂-C₂₈ heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C₂-C₂₈ alkenyl, and optionally substituted C₂-C₂₈ alkynyl; L is selected from the group consisting of a bond, optionally substituted C1-C6 alkylenyl, optionally substituted C₂-C₆ alkenylenyl, optionally substituted C₂-C₆ heteroalkylenyl, optionally substituted C3-C8 cycloalkylenyl, optionally substituted C2-C8 heterocycloalkylenyl, optionally substituted C₇-C₁₂ aralkylenyl, optionally substituted C₅-C₁₂ heteroaralkylenyl, optionally substituted C6-C10 arylenyl, and optionally substituted C2-C10 heteroarylenyl; and each occurrence of R^A and R^B is independently selected from the group consisting of H, C(=O)(optionally substituted C₁-C₆ alkyl), C(=O)(optionally substituted C₃-C₈ cycloalkyl), - 30 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) C(=O)(optionally substituted C2-C8 heterocycloalkyl), C(=O)(optionally substituted C6-C10 aryl), C(=O)(optionally substituted C₂-C₁₀ heteroaryl), optionally substituted C₁-C₆ alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted C₂-C₁₀ heteroaryl. In certain embodiments, R² is n-butyl. In certain embodiments, R² is H. In certain embodiments, R² is CH₂OCH₂CH₃. In certain embodiments, R² is CH₂NHCH₂CH₃. In certain embodiments, R^3a is H. In certain embodiments, R^3b is H. In certain embodiments, at least one of R^4a, R^4b, R^4c, and R^4d is H. In certain embodiments, at least two of R^4a, R^4b, R^4c, and R^4d are H. In certain embodiments, at least three of R^4a, R^4b, R^4c, and R^4d are H. In certain embodiments, each of R^4a, R^4b, R^4c, and R^4d are H. In certain embodiments, L is optionally substituted C₇-C₁₂ aralkylenyl. In certain embodiments, L . In certain embodiments, embodiments, R^6b is R^7a. In certain embodiments, R^6a is R^7b. In certain embodiments, R^6b is R^7b. In certain embodiments, R^6a is - CH₂CH₂C(=O)OR^7a. In certain embodiments, R^6b is -CH₂CH₂C(=O)OR^7a. In certain embodiments, R^7a is optionally substituted C6-C28 alkyl. In certain embodiments, R^7a is optionally substituted C₆-C₂₈ alkenyl. In certain embodiments, R^7a is optionally substituted C6-C28 heteroalkyl. In certain embodiments, R^7b is optionally substituted C₆-C₂₈ alkyl. In certain embodiments, R^7b is optionally substituted C6-C28 alkenyl. In certain embodiments, R^7b is optionally substituted C₆-C₂₈ heteroalkyl. In certain embodiments, R^6a is -CH2CH(OH)(optionally substituted C6-C28 alkyl). In certain embodiments, R^6a is CH₂CH(OH)(optionally substituted C₆-C₂₈ alkenyl). In certain embodiments, R^6a is -CH2CH(OH)(optionally substituted C6-C28 heteroalkyl). In certain embodiments, R^6a is -CH₂CH₂C(=O)O(optionally substituted C₆-C₂₈ alkyl). In certain (optionally substituted C6-C28 alkenyl). In certain (optionally substituted C₆-C₂₈ heteroalkyl). In -CH2CH(OH)(optionally substituted C6-C28 alkyl). In certain embodiments, R^6b is CH₂CH(OH)(optionally substituted C₆-C₂₈ alkenyl). In certain embodiments, R^6b is -CH2CH(OH)(optionally substituted C6-C28 heteroalkyl). In certain embodiments, R^6b is -CH2CH2C(=O)O(optionally substituted C6-C28 alkyl). In certain - 31 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) embodiments, R^6b is -CH2CH2C(=O)O(optionally substituted C6-C28 alkenyl). In certain embodiments, R^6b is -CH₂CH₂C(=O)O(optionally substituted C₆-C₂₈ heteroalkyl). In certain embodiments, R^6a is -CH2CH(OH)(CH2)9CH3. In certain embodiments, R^6a is -CH₂CH₂C(=O)O(CH₂)₁₁CH₃. In certain embodiments, R^6b is -CH2CH(OH)(CH2)9CH3. In certain embodiments, R^6b is -CH₂CH₂C(=O)O(CH₂)₁₁CH₃. In certain embodiments, the compound is: . In . Ionizable Lipids and/or Cationic Lipids The scope of ionizable lipids contemplated for use in the present disclosure is not limited to ionizable lipids exemplified herein. 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. Pat. Nos.5,208,036; 5,264,618; 5,279,833; 5,283,185; 5,753,613; and 5,785,992; and PCT Publication Nos. WO1996/010390, WO2021077066, WO2021077067, WO2023015200, WO2023056282, and WO2023056418, the disclosures of which are herein incorporated by reference in their entirety for all purposes. 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- - 32 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) dimethylaminopropane (DLinDMA), 1,2-dilinolenyloxy-N,N-dimethylaminopropane (DLenDMA), 2,2-dilinoleyl-4-(2-dimethylaminoethyl)-[1,3]-dioxolane (DLin-K-C2-DMA; "XTC2"), 2,2-dilinoleyl-4-(3- 45 dimethylaminopropyl)- 1,3]-dioxolane (D Lin-K-C3-D MA), 2,2-dilinoleyl-4-(4-dimethylaminobutyl)-[1,3]-dioxolane (DLin-K-C4-DMA), 2,2- dilinoleyl-5-dimethylaminomethyl-[1,3]-dioxane (DLin-K6-DMA), 2,2-dilinoleyl-4-N- methylpepiazino-[1,3]-dioxolane (DLin-K-MPZ), 2,2-dili-noleyl-4-dimethylaminomethyl- [1,3]-dioxolane (DLin-KDMA), 1,2-dilinoleylcarbamoyloxy-3-dimethylaminopropane (D Lin-C-DAP), 1,2-dilinoleyoxy-3-(dimethylaminoacetoxypropane (DLin-DAC), 1- 2dilinoleyoxy-3-morpholinopropane (DLin-MA), 1,2-dilinoleoyl-3-dimethylaminopropane (DLinDAP), 1,2-dilinoleylthio-3-dimethylaminopropane (DLin-2-DMAP), 1,2-dilinoleyloxy- 3-trimethylaminopropane chloride salt (DLin-TMA.Cl), 1,2-dilinoleoyl-3- trimethylaminopropane chloride salt (DLin-TAP.Cl), 1,2-dilinoleyloxy-3-(N- methylpiperazino)propane (D Lin-MPZ), 3-(N,N-dilinoleylamino)-1,2-propanediol (D LinAP), 3-(N,N-dioleylamino)-1,2-propanedio (DOAP), 1,2-dilinoleyloxo-3-(2-N,N- dimethylamino)ethoxypropane (D Lin-EG-D MA), N,N-dioleyl-N,N-dimethylanrmonium chloride (DODAC), 1,2-dioleyloxy-N,N-dimethylaminopropane (DODMA), 1,2- distearyloxy-N,N-dimethylaminopropane (DSD MA), N-(1-(2,3-dioleyloxy)propyl)-N,N,N- trimethylammonium chloride (DOTMA), N,N-distearyl-N,N-dimethylammonium bromide (DDAB), N-(1-(2,3-dioleoyloxy)propyl)-N,N, N-trimethylammonium chloride (DOTAP), 3- (N-(N',N'dimethylaminoethane)-carbamoyl)cholesterol (DC-Chol), N-(l,2- dimyristyloxyprop-3-yl)-N,N-dimethyl-N-hydroxyethyl anrmonium bromide (DMRIE), 2,3- dioleyloxy-N-[2 (spermine-carboxamidoethyl]-N,N-dimethy 1-1- propanaminiumtrifluoroacetate (DOSPA), dioctadecylamidoglycyl spermine (DOGS), 3- dimethylamino-2-(cholest-5-en-3-beta-oxybutan-4-oxy)-1-(cis,cis-9,12- octadecadienoxy)propane (CLinDMA), 2-[5'-(cholest-5-en-3-beta-oxy)-3'-oxapentoxy)-3- dimethyl-1-(cis,cis-9',1-2'-octadecadienoxy) propane (CpLinDMA), N,N-dimethyl-3,4- 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- - 33 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) DMA, DLin-K-C4-DMA, DLin-K6-DMA, and DLin-K-MPZ, as well as additional cationic lipids, is described in U.S. Application Publication No. US 2011/0256175, the disclosure of which is herein incorporated by reference in its entirety for all purposes. The synthesis of cationic lipids such as DLin-K-DMA, DLin-CDAP, DLin-DAC, DLin-MA, DLinDAP, DLin-S-DMA, DLin-2-DMAP, DLin-TMA.Cl, DLin-TAP.Cl, DLin-MPZ, DLinAP, DOAP, and DLin-EG-DMA, as well as additional cationic lipids, is described in PCT Application No. PCT/US08/88676, filed December 31, 2008, the disclosure of which is herein incorporated by reference in its entirety for all purposes. The synthesis of cationic lipids such as CLinDMA, as well as additional cationic lipids, is described in U.S. Patent Publication No. 20060240554, the disclosure of which is herein incorporated by reference in its entirety for all purposes. Non-cationic Lipids (Helper Lipids and/or Cholesterol or Derivatives Thereof) 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), - 34 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) dimyristoylphosphatidylethanolamine (DMPE), distearoylphosphatidylethanolamine (DSPE), monomethylphosphatidylethanolamine, dimethylphosphatidylethanolamine, dielaidoylphosphatidylethanolamine (DEPE), stearoyloleoylphosphatidylethanolamine (SOPE), lysophosphatidylcholine, dilinoleoylphosphatidylcholine, and mixtures thereof. Other diacylphosphatidylcholine and diacylphosphatidylethanolamine phospholipids can also be used. The acyl groups in these lipids can be, for example, acyl groups derived from fatty acids having C10-C24 carbon chains, e.g., lauroyl, myristoyl, palmitoyl, stearoyl, or oleoyl. Additional examples of non-cationic lipids include sterols such as cholesterol and derivatives thereof such as cholestanol, cholestanone, cholestenone, coprostanol, cholesteryl- 2'-hydroxyethyl ether, cholesteryl-4'-hydroxybutyl ether, and mixtures thereof. In certain embodiments, the phospholipid is DPPC, DSPC, or mixtures thereof. Polymer Conjugated Lipid 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 - 35 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) together with a CPL. The conjugated lipid that inhibits aggregation of particles may comprise a PEG-lipid including, e.g., a PEG-diacylglycerol (DAG), a PEG dialkyloxypropyl (DAA), a PEG-phospholipid, a PEG-ceramide (Cer), or mixtures thereof. The PEGDAA conjugate may be PEG-dilauryloxypropyl (C₁₂), a PEG-dimyristyloxypropyl (C₁₄), a PEG- dipalmityloxypropyl (C16), a PEG-distearyloxypropyl (C18), or mixtures thereof. Additional PEG-lipid conjugates suitable for use in the disclosure include, but are not limited to, mPEG2000-l,2-diO-alkyl-sn3-carbomoylglyceride (PEG-C-DOMG). The synthesis of PEG-C-DOMG is described in PCT Application No. PCT/US08/88676, filed December 31, 2008, the disclosure of which is herein incorporated by reference in its entirety for all purposes. Yet additional PEG-lipid conjugates suitable for use in the disclosure include, without limitation, l-[8'-(l,2-dimyristoyl-3-propanoxy)-carboxamido-3',6'- dioxaoctanyl] carbamoyl-methyl-poly(ethylene glycol) (2 KPEG-DMG). The synthesis of 2 KPEG-DMG is described in U.S. Patent No.7,404,969, the disclosure of which is herein incorporated by reference in its entirety for all purposes. The PEG moiety of the PEG-lipid conjugates described herein may comprise an average molecular weight ranging from about 550 daltons to about 10,000 daltons. In certain instances, the PEG moiety has an average molecular weight of from about 750 daltons to about 5,000 daltons (e.g., from about 1,000 daltons to about 5,000 daltons, from about 1,500 daltons to about 3,000 daltons, from about 750 daltons to about 3,000 daltons, from about 750 daltons to about 2,000 daltons, etc.). In some embodiments, the PEG moiety has an average molecular weight of about 2,000 daltons or about 750 daltons. In addition to the foregoing, it will be readily apparent to those of skill in the art that other hydrophilic polymers can be used in place of PEG. Examples of suitable polymers that can be used in place of PEG include, but are not limited to, polyvinylpyrrolidone, polymethyloxazoline, polyethyloxazoline, polyhydroxypropyl methacrylamide, polymethacrylamide and polydimethylacrylamide, polylactic acid, polyglycolic acid, and 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., - 36 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) PEG-lipid conjugate) may comprise from about 0.1 mol% to about 2 mol%, from about 0.5 mol% to about 2 mol%, from about 1 mol% to about 2 mol%, from about 0.6 mol% to about 1.9 mol%, from about 0.7 mol% to about 1.8 mol%, from about 0.8 mol% to about 1.7 mol%, from about 1 mol% to about 1.8 mol%, from about 1.2 mol% to about 1.8 mol%, from about 1.2 mol% to about 1.7 mol%, from about 1.3 mol% to about 1.6 mol%, from about 1.4 mol% to about 1.5 mol%, or about 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, or 2 mol% (or any fraction thereof or range therein) of the total lipid present in the particle. In the lipid nanoparticles of the present disclosure, the active agent or therapeutic agent may be fully encapsulated within the lipid portion of the particle, thereby protecting the active agent or therapeutic agent from enzymatic degradation. In some embodiments, a nucleic acid-lipid particle comprising a nucleic acid such as a messenger RNA (i.e., mRNA) is fully encapsulated within the lipid portion of the particle, thereby protecting the nucleic acid from nuclease degradation. In certain instances, the nucleic acid in the nucleic acid-lipid particle is not substantially degraded after exposure of the particle to a nuclease at 37° C. for at least about 20, 30, 45, or 60 minutes. In certain other instances, the nucleic acid in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C. for at least about 30, 45, or 60 minutes or at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, or 36 hours. In other embodiments, the active agent or therapeutic agent (e.g., nucleic acid such as siRNA) is complexed with the lipid portion of the particle. One of the benefits of the formulations of the present disclosure is that the lipid particle compositions are substantially non-toxic to mammals such as humans. Lipid Nanoparticle (LNP) Compositions In one aspect, the present disclosure provides a lipid nanoparticle (LNP). In certain embodiments, the LNP comprises at least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent. In certain embodiments, the LNP comprises at least one ionizable lipid. In certain embodiments, the LNP comprises at least one helper lipid. In certain embodiments, the LNP comprises cholesterol. In certain embodiments, the LNP comprises at least one polymer conjugated lipid. In certain embodiments, the LNP comprises (a) at least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent. In certain embodiments, the LNP comprises (b) at least one ionizable lipid. In certain embodiments, the LNP comprises (c) at least one helper lipid. In certain embodiments, the LNP comprises (d) cholesterol. - 37 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) In certain embodiments, the LNP comprises (e) at least one polymer conjugated lipid. In certain embodiments, the LNP comprises the hydrocarbyl substituted TLR agonist is a compound of Formula (II): A(B)_n (II), wherein: A is a toll-like receptor (TLR) agonist; each occurrence of B is independently selected from the group consisting of R^6a and ; of R^6a and R^6b is independently selected from the group consisting of - (optionally substituted C1-C6 alkylenyl)-C(=O)OR^7a, -(optionally substituted C1-C6 alkylenyl)-C(=O)N(R^7a)(R^7b), -(optionally substituted C₁-C₆ alkylenyl)-C(=O)R^7a, - (optionally substituted C1-C6 alkylenyl)-(R^7a), -C(=O)OR^7a, -C(=O)N(R^7a)(R^7b), -C(=O)R^7a, and R^7a, each occurrence of R^7a and R^7b is independently selected from the group consisting of optionally substituted C1-C28 alkyl, optionally substituted C2-C28 heteroalkyl, optionally substituted C₃-C₈ cycloalkyl, optionally substituted C₂-C₈ heterocycloalkyl, optionally substituted C2-C28 alkenyl, and optionally substituted C2-C28 alkynyl; L is bond, optionally substituted C₁-C₆ alkylenyl, optionally substituted C₂-C₆ alkenylenyl, optionally substituted C2-C6 heteroalkylenyl, optionally substituted C3-C8 cycloalkylenyl, optionally substituted C₂-C₈ heterocycloalkylenyl, optionally substituted C₇- C12 aralkylenyl, optionally substituted C5-C12 heteroaralkylenyl, optionally substituted C6-C10 arylenyl, and optionally substituted C₂-C₁₀ heteroarylenyl; and n is an integer selected from the group consisting of 1, 2, 3, and 4. In certain embodiments, the TLR agonist is 1-(4-(aminomethyl)benzyl)-2-butyl-1H- imidazo[4,5-c]quinolin-4-amine (TLR7/8 agonist 1). In certain embodiments, the TLR agonist is 1-isobutyl-1H-imidazo[4,5-c]quinolin-4-amine (imiquimod). In certain embodiments, the TLR agonist is 1-(4-amino-2-(ethoxymethyl)-1H-imidazo[4,5-c]quinolin- 1-yl)-2-methylpropan-2-ol (resiquimod). In certain embodiments, the TLR agonist is 1-(4- amino-2-((ethylamino)methyl)-1H-imidazo[4,5-c]quinolin-1-yl)-2-methylpropan-2-ol (gardiquimod). In certain embodiments, the least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent is the compound of Formula (I), or a salt, solvate, - 38 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) stereoisomer, or isotopologue thereof: , wherein: R¹ is selected from the group substituted C1-C6 alkyl, optionally substituted C₃-C₈ cycloalkyl, optionally substituted C₂-C₈ heterocycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted heteroaryl; R² is selected from the group consisting of H, R⁵, optionally substituted C₁-C₆ alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C₆-C₁₀ aryl, and optionally substituted heteroaryl; R^3a and R^3b are each independently selected from the group consisting of H, R⁵, R^6a, and optionally substituted C₁-C₆ alkyl; R^4a, R^4b, R^4c, and R^4d are each independently selected from the group consisting of H, R⁵, halogen, CN, NO₂, optionally substituted C₁-C₆ alkyl, optionally substituted C₃-C₈ cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C6-C10 aryl, optionally substituted C₂-C₁₀ heteroaryl, OR^A, N(R^A)(R^B), C(=O)N(R^A)(R^B), C(=O)R^A, C(=O)OR^A, OC(=O)R^A, OC(=O)OR^A, SR^A, S(=O)R^A, S(=O)2R^A, N(R^A)S(=O)2R^B, wherein at least one of R¹, R², R^3a, R^3b, R^4a, R^4b, R^4c, and R^4d is R⁵, or at least one of R^3a and R^3b is R^6a; each occurrence of R^6a and R^6b is independently selected from the group consisting of - (optionally substituted C1-C6 alkylenyl)-C(=O)OR^7a, -(optionally substituted C1-C6 alkylenyl)-C(=O)N(R^7a)(R^7b), -(optionally substituted C₁-C₆ alkylenyl)-C(=O)R^7a, - (optionally substituted C1-C6 alkylenyl)-(R^7a), -C(=O)OR^7a, -C(=O)N(R^7a)(R^7b), -C(=O)R^7a, and R^7a; each occurrence of R^7a and R^7b is independently selected from the group consisting of optionally substituted C₆-C₂₈ alkyl, optionally substituted C₆-C₂₈ heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C6-C8 heterocycloalkyl, optionally - 39 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) substituted C6-C28 alkenyl, and optionally substituted C6-C28 alkynyl; L is selected from the group consisting of a bond, optionally substituted C₁-C₆ alkylenyl, optionally substituted C2-C6 alkenylenyl, optionally substituted C2-C6 heteroalkylenyl, optionally substituted C₃-C₈ cycloalkylenyl, optionally substituted C₂-C₈ heterocycloalkylenyl, optionally substituted C7-C12 aralkylenyl, optionally substituted C5-C12 heteroaralkylenyl, optionally substituted C₆-C₁₀ arylenyl, and optionally substituted C₂-C₁₀ heteroarylenyl; and each occurrence of R^A and R^B is independently selected from the group consisting of H, C(=O)(optionally substituted C1-C6 alkyl), C(=O)(optionally substituted C3-C8 cycloalkyl), C(=O)(optionally substituted C₂-C₈ heterocycloalkyl), C(=O)(optionally substituted C₆-C₁₀ aryl), C(=O)(optionally substituted C2-C10 heteroaryl), optionally substituted C1-C6 alkyl, optionally substituted C₃-C₈ cycloalkyl, optionally substituted C₆-C₁₀ aryl, and optionally substituted C2-C10 heteroaryl. In certain embodiments, exactly one of R¹, R², R^3a, R^3b, R^4a, R^4b, R^4c, and R^4d is R⁵. In certain embodiments, R¹ is R⁵. In certain embodiments, R¹ is CH2CH(CH3)2. In certain embodiments, R¹ is CH2C(OH)(CH3)2. In certain embodiments, the compound of Formula (I) is a compound of Formula (Ia), or a salt, solvate, stereoisomer, or isotopologue thereof: R² is selected from the C1-C6 alkyl, optionally substituted C₃-C₈ cycloalkyl, optionally substituted C₂-C₈ heterocycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted heteroaryl; R^3a and R^3b are each independently selected from the group consisting of H, R^6a, and optionally substituted C1-C6 alkyl; R^4a, R^4b, R^4c, and R^4d are each independently selected from the group consisting of H, halogen, CN, NO2, optionally substituted C1-C6 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C₂-C₈ heterocycloalkyl, optionally substituted C₆-C₁₀ aryl, optionally substituted C2-C10 heteroaryl, OR^A, N(R^A)(R^B), C(=O)N(R^A)(R^B), C(=O)R^A, C(=O)OR^A, OC(=O)R^A, OC(=O)OR^A, SR^A, S(=O)R^A, S(=O)₂R^A, N(R^A)S(=O)₂R^B, - 40 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) N(R^A)C(=O)R^B, and S(=O)2N(R^A)(R^B); each occurrence of R^6a and R^6b is independently selected from the group consisting of - (optionally substituted C1-C6 alkylenyl)-C(=O)OR^7a, -(optionally substituted C1-C6 alkylenyl)-C(=O)N(R^7a)(R^7b), -(optionally substituted C₁-C₆ alkylenyl)-C(=O)R^7a, - (optionally substituted C1-C6 alkylenyl)-(R^7a), -C(=O)OR^7a, -C(=O)N(R^7a)(R^7b), -C(=O)R^7a, and R^7a; each occurrence of R^7a and R^7b is independently selected from the group consisting of optionally substituted C₁-C₂₈ alkyl, optionally substituted C₂-C₂₈ heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C₂-C₂₈ alkenyl, and optionally substituted C₂-C₂₈ alkynyl; L is selected from the group consisting of a bond, optionally substituted C1-C6 alkylenyl, optionally substituted C₂-C₆ alkenylenyl, optionally substituted C₂-C₆ heteroalkylenyl, optionally substituted C3-C8 cycloalkylenyl, optionally substituted C2-C8 heterocycloalkylenyl, optionally substituted C₇-C₁₂ aralkylenyl, optionally substituted C₅-C₁₂ heteroaralkylenyl, optionally substituted C6-C10 arylenyl, and optionally substituted C2-C10 heteroarylenyl; and each occurrence of R^A and R^B is independently selected from the group consisting of H, C(=O)(optionally substituted C1-C6 alkyl), C(=O)(optionally substituted C3-C8 cycloalkyl), C(=O)(optionally substituted C₂-C₈ heterocycloalkyl), C(=O)(optionally substituted C₆-C₁₀ aryl), C(=O)(optionally substituted C2-C10 heteroaryl), optionally substituted C1-C6 alkyl, optionally substituted C₃-C₈ cycloalkyl, optionally substituted C₆-C₁₀ aryl, and optionally substituted C2-C10 heteroaryl. In certain embodiments, R² is n-butyl. In certain embodiments, R² is H. In certain embodiments, R² is CH2OCH2CH3. In certain embodiments, R² is CH2NHCH2CH3. In certain embodiments, R^3a is H. In certain embodiments, R^3b is H. In certain embodiments, at least one of R^4a, R^4b, R^4c, and R^4d is H. In certain embodiments, at least two of R^4a, R^4b, R^4c, and R^4d are H. In certain embodiments, at least three of R^4a, R^4b, R^4c, and R^4d are H. In certain embodiments, each of R^4a, R^4b, R^4c, and R^4d are H. In certain embodiments, L is optionally substituted C7-C12 aralkylenyl. In certain embodiments, L . In certain embodiments, is In certain embodiments, R^6b is R^7a. In certain - 41 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) embodiments, R^6a is R^7b. In certain embodiments, R^6b is R^7b. In certain embodiments, R^6a is - CH₂CH₂C(=O)OR^7a. In certain embodiments, R^6b is -CH₂CH₂C(=O)OR^7a. In certain embodiments, R^7a is optionally substituted C6-C28 alkyl. In certain embodiments, R^7a is optionally substituted C₆-C₂₈ alkenyl. In certain embodiments, R^7a is optionally substituted C6-C28 heteroalkyl. In certain embodiments, R^7b is optionally substituted C₆-C₂₈ alkyl. In certain embodiments, R^7b is optionally substituted C6-C28 alkenyl. In certain embodiments, R^7b is optionally substituted C₆-C₂₈ heteroalkyl. In certain embodiments, R^6a is -CH2CH(OH)(optionally substituted C6-C28 alkyl). In certain embodiments, R^6a is CH₂CH(OH)(optionally substituted C₆-C₂₈ alkenyl). In certain embodiments, R^6a is -CH2CH(OH)(optionally substituted C6-C28 heteroalkyl). In certain embodiments, R^6a is -CH₂CH₂C(=O)O(optionally substituted C₆-C₂₈ alkyl). In certain embodiments, R^6a is -CH2CH2C(=O)O(optionally substituted C6-C28 alkenyl). In certain embodiments, R^6a is -CH₂CH₂C(=O)O(optionally substituted C₆-C₂₈ heteroalkyl). In certain embodiments, R^6b is -CH2CH(OH)(optionally substituted C6-C28 alkyl). In certain embodiments, R^6b is CH2CH(OH)(optionally substituted C6-C28 alkenyl). In certain embodiments, R^6b is -CH2CH(OH)(optionally substituted C6-C28 heteroalkyl). In certain embodiments, R^6b is -CH2CH2C(=O)O(optionally substituted C6-C28 alkyl). In certain embodiments, R^6b is -CH₂CH₂C(=O)O(optionally substituted C₆-C₂₈ alkenyl). In certain embodiments, R^6b is -CH2CH2C(=O)O(optionally substituted C6-C28 heteroalkyl). In certain embodiments, R^6a is -CH₂CH(OH)(CH₂)₉CH₃. In certain embodiments, R^6a is -CH2CH2C(=O)O(CH2)11CH3. In certain embodiments, R^6b is -CH₂CH(OH)(CH₂)₉CH₃. In certain embodiments, R^6b is -CH2CH2C(=O)O(CH2)11CH3. In certain embodiments, the compound is: . In - 42 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) . , wherein: R^8a, R^8b, R^8c, R^8d, R^8e, and R^8f are each independently selected from the group consisting of -(optionally substituted C1-C6 alkylenyl)-C(=O)OR^9a, -(optionally substituted C1-C6 alkylenyl)-C(=O)N(R^9a)(R^9b), -(optionally substituted C₁-C₆ alkylenyl)-C(=O)R^9a, - (optionally substituted C1-C6 alkylenyl)-(R^9a), -C(=O)OR^9a, -C(=O)N(R^9a)(R^9b), -C(=O)R^9a, and R^9a; and each occurrence of R^9a and R^9b is independently selected from the group consisting of optionally substituted C1-C28 alkyl, optionally substituted C2-C28 heteroalkyl, optionally substituted C₃-C₈ cycloalkyl, optionally substituted C₂-C₈ heterocycloalkyl, optionally substituted C2-C28 alkenyl, and optionally substituted C2-C28 alkynyl. In certain embodiments, R^8a is R^9a. In certain embodiments, R^8a is R^9b. In certain embodiments, R^8a is -CH2CH2C(=O)OR^9a. In certain embodiments, R^8a is - CH₂CH₂C(=O)OR^9b. In certain embodiments, R^8b is R^9a. In certain embodiments, R^8b is R^9b. In certain embodiments, R^8b is -CH2CH2C(=O)OR^9a. In certain embodiments, R^8b is - CH₂CH₂C(=O)OR^9b. In certain embodiments, R^8c is R^9a. In certain embodiments, R^8c is R^9b. In certain embodiments, R^8c is -CH2CH2C(=O)OR^9a. In certain embodiments, R^8c is - - 43 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) CH2CH2C(=O)OR^9b. In certain embodiments, R^8d is R^9a. In certain embodiments, R^8d is R^9b. In certain embodiments, R^8d is -CH₂CH₂C(=O)OR^9a. In certain embodiments, R^8d is - CH2CH2C(=O)OR^9b. In certain embodiments, R^8e is R^9a. In certain embodiments, R^8e is R^9b. In certain embodiments, R^8e is -CH₂CH₂C(=O)OR^9a. In certain embodiments, R^8e is - CH2CH2C(=O)OR^9b. In certain embodiments, R^8f is R^9a. In certain embodiments, R^8f is R^9b. In certain embodiments, R^8f is -CH₂CH₂C(=O)OR^9a. In certain embodiments, R^8f is - CH2CH2C(=O)OR^9b. In certain embodiments, R^9a is optionally substituted C₁-C₂₈ alkyl. In certain embodiments, R^9a is optionally substituted C1-C28 alkenyl. In certain embodiments, R^9a is optionally substituted C₁-C₂₈ heteroalkyl. In certain embodiments, R^9b is optionally substituted C1-C28 alkyl. In certain embodiments, R^9b is optionally substituted C1-C28 alkenyl. In certain embodiments, R^9b is optionally substituted C₁-C₂₈ heteroalkyl. In certain embodiments, R^7a is -CH2CH(OH)(optionally substituted C1-C28 alkyl). In certain embodiments, R^7a is -CH₂CH(OH)(optionally substituted C₁-C₂₈ alkenyl). In certain embodiments, R^7a is -CH2CH(OH)(optionally substituted C2-C28 heteroalkyl). In certain embodiments, R^7a is -CH2CH2C(=O)O(optionally substituted C1-C28 alkyl). In certain (optionally substituted C1-C28 alkenyl). In certain (optionally substituted C2-C28 heteroalkyl). In certain substituted C₁-C₂₈ alkyl). In certain embodiments, R^7b is -CH2CH(OH)(optionally substituted C1-C28 alkenyl). In certain embodiments, R^7b is -CH₂CH(OH)(optionally substituted C₂-C₂₈ heteroalkyl). In certain embodiments, R^7b is -CH2CH2C(=O)O(optionally substituted C1-C28 alkyl). In certain embodiments, R^7b is -CH₂CH₂C(=O)O(optionally substituted C₁-C₂₈ alkenyl). In certain embodiments, R^7b is -CH2CH2C(=O)O(optionally substituted C2-C28 heteroalkyl). In certain embodiments, R^7a is -CH₂CH(OH)(CH₂)₉CH₃. In certain embodiments, R^7a is -CH2CH2C(=O)O(CH2)11CH3. In certain embodiments, R^7b is -CH2CH(OH)(CH2)9CH3. In certain embodiments, R^7b is -CH₂CH₂C(=O)O(CH₂)₁₁CH₃. In certain embodiments, the at least one ionizable lipid comprises: . - 44 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) with at least one hydrocarbyl substituent comprises about 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 10.5, 11.0, 11.5, 12.0, 12.5, 13.0, 13.5, 14.0, 14.5, 15.0, 15.5, 16.0, 16.5, 17.0, or about 17.5 mol% of the LNP. In certain embodiments the at least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent comprises less than about 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 10.5, 11.0, 11.5, 12.0, 12.5, 13.0, 13.5, 14.0, 14.5, 15.0, 15.5, 16.0, 16.5, 17.0, or about 17.5 mol% of the LNP. In certain embodiments the at least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent comprises about 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 10.5, 11.0, 11.5, 12.0, 12.5, 13.0, 13.5, 14.0, 14.5, 15.0, 15.5, 16.0, 16.5, 17.0, or about 17.5 mol% of the LNP. In certain embodiments the at least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent comprises about 1 mol% of the LNP. In certain embodiments the at least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent comprises about 1.5 mol% of the LNP. In certain embodiments the at least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent comprises about 5 mol% of the LNP. In certain embodiments the at least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent comprises about 10 mol% of the LNP. In certain embodiments the at least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent comprises about 17.5 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, 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 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, - 45 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) 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 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, or about 60 mol% of the LNP. In certain embodiments, the at least one ionizable lipid comprises more than about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or about 60 mol% of the LNP. In certain embodiments, the at least one ionizable lipid comprises more than about 34 mol% of the LNP. In certain embodiments, the at least one ionizable lipid comprises more than about 32.5 mol% of the LNP. In certain embodiments, the at least one ionizable lipid comprises more than about 30 mol% of the LNP. In certain embodiments, the at least one ionizable lipid comprises more than about 25 mol% of the LNP. In certain embodiments, the at least one ionizable lipid comprises more than about 17.5 mol% of the LNP. In certain embodiments, the at least one helper lipid comprises dioleoylphosphatidylethanolamine (DOPE). In certain embodiments, the at least one helper lipid comprises distearoylphosphatidylcholine (DSPC). 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, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or about 50 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, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or about 50 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, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or about 50 mol% of the LNP. In certain embodiments, the at least one helper lipid comprises about 16 mol% of the LNP. In certain embodiments, cholesterol comprises about 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, or about 60 mol% - 46 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) of the LNP. In certain embodiments, cholesterol comprises less than about 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, or about 60 mol% of the LNP. In certain embodiments, cholesterol comprises more than about 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, or about 60 mol% of the LNP. In certain embodiments, the cholesterol comprises about 46.5 mol% of the LNP. In certain embodiments, the at least one polymer conjugated lipid comprises 1,2- dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (DMG-PEG 2000). 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, 9.8, 9.9, 10.0, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11.0, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12.0, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13.0, 13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14.0, 14.1, 14.2, 14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, 15.0, 15.1, 15.2, 15.3, 15.4, 15.5, 15.6, 15.7, 15.8, 15.9, 16.0, 16.1, 16.2, 16.3, 16.4, 16.5, 16.6, 16.7, 16.8, 16.9, 17.0, 17.1, 17.2, 17.3, 17.4, 17.5, 17.6, 17.7, 17.8, 17.9, 18.0, 18.1, 18.2, 18.3, 18.4, 18.5, 18.6, 18.7, 18.8, 18.9, 19.0, 19.1, 19.2, 19.3, 19.4, 19.5, 19.6, 19.7, 19.8, 19.9, or about 20.0 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, 9.8, 9.9, 10.0, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11.0, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12.0, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13.0, 13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14.0, 14.1, 14.2, 14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, 15.0, 15.1, - 47 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) 15.2, 15.3, 15.4, 15.5, 15.6, 15.7, 15.8, 15.9, 16.0, 16.1, 16.2, 16.3, 16.4, 16.5, 16.6, 16.7, 16.8, 16.9, 17.0, 17.1, 17.2, 17.3, 17.4, 17.5, 17.6, 17.7, 17.8, 17.9, 18.0, 18.1, 18.2, 18.3, 18.4, 18.5, 18.6, 18.7, 18.8, 18.9, 19.0, 19.1, 19.2, 19.3, 19.4, 19.5, 19.6, 19.7, 19.8, 19.9, or about 20.0 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, 9.8, 9.9, 10.0, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11.0, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12.0, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13.0, 13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14.0, 14.1, 14.2, 14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, 15.0, 15.1, 15.2, 15.3, 15.4, 15.5, 15.6, 15.7, 15.8, 15.9, 16.0, 16.1, 16.2, 16.3, 16.4, 16.5, 16.6, 16.7, 16.8, 16.9, 17.0, 17.1, 17.2, 17.3, 17.4, 17.5, 17.6, 17.7, 17.8, 17.9, 18.0, 18.1, 18.2, 18.3, 18.4, 18.5, 18.6, 18.7, 18.8, 18.9, 19.0, 19.1, 19.2, 19.3, 19.4, 19.5, 19.6, 19.7, 19.8, 19.9, or about 20.0 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 LNP has a molar ratio of (a) : (b) : (c) : (d) : (e) of about 5 : 30 : 16 : 46.5 : 2.5. In certain embodiments, the LNP further comprises at least one cargo molecule. In certain embodiments, the cargo 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 is a nucleic acid. In certain embodiments, the nucleic acid is DNA or RNA. In certain embodiments, the nucleic acid is selected from the group consisting of mRNA, cDNA, pDNA, microRNA, 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 cargo is mRNA. In certain embodiments, the LNP has a weight ratio of total lipidoid (i.e., hydrocarbyl substituted toll-like receptor agonist and ionizable lipid) to mRNA ranging from about 5:1, - 48 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, or about 20:1 (i.e., weight ratio of (a) + (b) : mRNA). In certain embodiments, the LNP has a weight ratio of total lipidoid (i.e., hydrocarbyl substituted toll-like receptor agonist and ionizable lipid) to mRNA of about 10:1 (i.e., weight ratio of (a) + (b) : mRNA). In certain embodiments, the mRNA encodes SARS-CoV-2, an immunogenic fragment thereof (e.g., spike protein), or a modified derivative thereof. Methods In one aspect, the present disclosure provides a method of generating an innate immune response in a subject, the method comprising administering to the subject the lipid nanoparticle (LNP) of the present disclosure or a pharmaceutical composition of the present disclosure. In another aspect, the present disclosure provides a method of treating, preventing, and/or ameliorating an infection, disease, or disorder in a subject, the method comprising administering to the subject a lipid nanoparticle (LNP). In certain embodiments, the LNP comprises (a) at least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent. In certain embodiments, the LNP comprises (b) at least one ionizable lipid. In certain embodiments, the LNP comprises (c) at least one helper lipid. In certain embodiments, the LNP comprises (d) cholesterol. In certain embodiments, the LNP comprises (e) at least one polymer conjugated lipid. In certain embodiments, the LNP comprises (f) at least one cargo molecule. In certain embodiments, the LNP comprises the hydrocarbyl substituted TLR agonist is a compound of Formula (II): A(B)_n (II), wherein: A is a toll-like receptor (TLR) agonist; each occurrence of B is independently selected from the group consisting of R^6a and ; each occurrence of R^6a and R^6b is independently selected from the group consisting of - (optionally substituted C1-C6 alkylenyl)-C(=O)OR^7a, -(optionally substituted C1-C6 alkylenyl)-C(=O)N(R^7a)(R^7b), -(optionally substituted C1-C6 alkylenyl)-C(=O)R^7a, - - 49 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) (optionally substituted C1-C6 alkylenyl)-(R^7a), -C(=O)OR^7a, -C(=O)N(R^7a)(R^7b), -C(=O)R^7a, and R^7a, each occurrence of R^7a and R^7b is independently selected from the group consisting of optionally substituted C₁-C₂₈ alkyl, optionally substituted C₂-C₂₈ heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C₂-C₂₈ alkenyl, and optionally substituted C₂-C₂₈ alkynyl; L is bond, optionally substituted C1-C6 alkylenyl, optionally substituted C2-C6 alkenylenyl, optionally substituted C₂-C₆ heteroalkylenyl, optionally substituted C₃-C₈ cycloalkylenyl, optionally substituted C2-C8 heterocycloalkylenyl, optionally substituted C7- C₁₂ aralkylenyl, optionally substituted C₅-C₁₂ heteroaralkylenyl, optionally substituted C₆-C₁₀ arylenyl, and optionally substituted C2-C10 heteroarylenyl; and n is an integer selected from the group consisting of 1, 2, 3, and 4. In certain embodiments, the TLR agonist is 1-(4-(aminomethyl)benzyl)-2-butyl-1H- imidazo[4,5-c]quinolin-4-amine (TLR7/8 agonist 1). In certain embodiments, the TLR agonist is 1-isobutyl-1H-imidazo[4,5-c]quinolin-4-amine (imiquimod). In certain embodiments, the TLR agonist is 1-(4-amino-2-(ethoxymethyl)-1H-imidazo[4,5-c]quinolin- 1-yl)-2-methylpropan-2-ol (resiquimod). In certain embodiments, the TLR agonist is 1-(4- amino-2-((ethylamino)methyl)-1H-imidazo[4,5-c]quinolin-1-yl)-2-methylpropan-2-ol (gardiquimod). In certain embodiments, the least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent is the compound of Formula (I), or a salt, solvate, stereoisomer, or isotopologue thereof: , wherein: R¹ is selected from the group consisting of H, R⁵, optionally substituted C1-C6 alkyl, optionally substituted C₃-C₈ cycloalkyl, optionally substituted C₂-C₈ heterocycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted heteroaryl; R² is selected from the group consisting of H, R⁵, optionally substituted C₁-C₆ alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C₆-C₁₀ aryl, and optionally substituted heteroaryl; - 50 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) R^3a and R^3b are each independently selected from the group consisting of H, R⁵, R^6a, and optionally substituted C₁-C₆ alkyl; R^4a, R^4b, R^4c, and R^4d are each independently selected from the group consisting of H, R⁵, halogen, CN, NO₂, optionally substituted C₁-C₆ alkyl, optionally substituted C₃-C₈ cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C6-C10 aryl, optionally substituted C₂-C₁₀ heteroaryl, OR^A, N(R^A)(R^B), C(=O)N(R^A)(R^B), C(=O)R^A, C(=O)OR^A, OC(=O)R^A, OC(=O)OR^A, SR^A, S(=O)R^A, S(=O)2R^A, N(R^A)S(=O)2R^B, N C and S ₂N R⁵ is , wherein at least one of R¹, R², R^3a, R^3b, R^4a, R^4b, R^4c, and R^4d is R⁵, or at least one of R^3a and R^3b is R^6a; each occurrence of R^6a and R^6b is independently selected from the group consisting of - (optionally substituted C₁-C₆ alkylenyl)-C(=O)OR^7a, -(optionally substituted C₁-C₆ alkylenyl)-C(=O)N(R^7a)(R^7b), -(optionally substituted C1-C6 alkylenyl)-C(=O)R^7a, - (optionally substituted C1-C6 alkylenyl)-(R^7a), -C(=O)OR^7a, -C(=O)N(R^7a)(R^7b), -C(=O)R^7a, and R^7a; each occurrence of R^7a and R^7b is independently selected from the group consisting of optionally substituted C₆-C₂₈ alkyl, optionally substituted C₆-C₂₈ heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C6-C8 heterocycloalkyl, optionally substituted C₆-C₂₈ alkenyl, and optionally substituted C₆-C₂₈ alkynyl; L is selected from the group consisting of a bond, optionally substituted C1-C6 alkylenyl, optionally substituted C₂-C₆ alkenylenyl, optionally substituted C₂-C₆ heteroalkylenyl, optionally substituted C3-C8 cycloalkylenyl, optionally substituted C2-C8 heterocycloalkylenyl, optionally substituted C₇-C₁₂ aralkylenyl, optionally substituted C₅-C₁₂ heteroaralkylenyl, optionally substituted C6-C10 arylenyl, and optionally substituted C2-C10 heteroarylenyl; and each occurrence of R^A and R^B is independently selected from the group consisting of H, C(=O)(optionally substituted C₁-C₆ alkyl), C(=O)(optionally substituted C₃-C₈ cycloalkyl), C(=O)(optionally substituted C2-C8 heterocycloalkyl), C(=O)(optionally substituted C6-C10 aryl), C(=O)(optionally substituted C₂-C₁₀ heteroaryl), optionally substituted C₁-C₆ alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted C2-C10 heteroaryl. - 51 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) In certain embodiments, exactly one of R¹, R², R^3a, R^3b, R^4a, R^4b, R^4c, and R^4d is R⁵. In certain embodiments, R¹ is R⁵. In certain embodiments, R¹ is CH₂CH(CH₃)₂. In certain embodiments, R¹ is CH2C(OH)(CH3)2. In certain embodiments, the compound of Formula (I) is a compound of Formula (Ia), or a salt, solvate, stereoisomer, or isotopologue thereof: R² is selected from the C1-C6 alkyl, optionally substituted C₃-C₈ C₂-C₈ heterocycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted heteroaryl; R^3a and R^3b are each independently selected from the group consisting of H, R^6a, and optionally substituted C1-C6 alkyl; R^4a, R^4b, R^4c, and R^4d are each independently selected from the group consisting of H, halogen, CN, NO2, optionally substituted C1-C6 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C₂-C₈ heterocycloalkyl, optionally substituted C₆-C₁₀ aryl, optionally substituted C2-C10 heteroaryl, OR^A, N(R^A)(R^B), C(=O)N(R^A)(R^B), C(=O)R^A, C(=O)OR^A, OC(=O)R^A, OC(=O)OR^A, SR^A, S(=O)R^A, S(=O)₂R^A, N(R^A)S(=O)₂R^B, N(R^A)C(=O)R^B, and S(=O)2N(R^A)(R^B); each occurrence of R^6a and R^6b is independently selected from the group consisting of - (optionally substituted C₁-C₆ alkylenyl)-C(=O)OR^7a, -(optionally substituted C₁-C₆ alkylenyl)-C(=O)N(R^7a)(R^7b), -(optionally substituted C1-C6 alkylenyl)-C(=O)R^7a, - (optionally substituted C₁-C₆ alkylenyl)-(R^7a), -C(=O)OR^7a, -C(=O)N(R^7a)(R^7b), -C(=O)R^7a, and R^7a; each occurrence of R^7a and R^7b is independently selected from the group consisting of optionally substituted C1-C28 alkyl, optionally substituted C2-C28 heteroalkyl, optionally substituted C₃-C₈ cycloalkyl, optionally substituted C₂-C₈ heterocycloalkyl, optionally substituted C2-C28 alkenyl, and optionally substituted C2-C28 alkynyl; L is selected from the group consisting of a bond, optionally substituted C₁-C₆ alkylenyl, optionally substituted C2-C6 alkenylenyl, optionally substituted C2-C6 heteroalkylenyl, optionally substituted C₃-C₈ cycloalkylenyl, optionally substituted C₂-C₈ - 52 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) heterocycloalkylenyl, optionally substituted C7-C12 aralkylenyl, optionally substituted C5-C12 heteroaralkylenyl, optionally substituted C₆-C₁₀ arylenyl, and optionally substituted C₂-C₁₀ heteroarylenyl; and each occurrence of R^A and R^B is independently selected from the group consisting of H, C(=O)(optionally substituted C1-C6 alkyl), C(=O)(optionally substituted C3-C8 cycloalkyl), C(=O)(optionally substituted C₂-C₈ heterocycloalkyl), C(=O)(optionally substituted C₆-C₁₀ aryl), C(=O)(optionally substituted C2-C10 heteroaryl), optionally substituted C1-C6 alkyl, optionally substituted C₃-C₈ cycloalkyl, optionally substituted C₆-C₁₀ aryl, and optionally substituted C2-C10 heteroaryl. In certain embodiments, R² is n-butyl. In certain embodiments, R² is H. In certain embodiments, R² is CH2OCH2CH3. In certain embodiments, R² is CH2NHCH2CH3. In certain embodiments, R^3a is H. In certain embodiments, R^3b is H. In certain embodiments, at least one of R^4a, R^4b, R^4c, and R^4d is H. In certain embodiments, at least two of R^4a, R^4b, R^4c, and R^4d are H. In certain embodiments, at least three of R^4a, R^4b, R^4c, and R^4d are H. In certain embodiments, each of R^4a, R^4b, R^4c, and R^4d are H. In certain embodiments, L is optionally substituted C7-C12 aralkylenyl. In certain embodiments, L . In certain embodiments, embodiments, R^6b is R^7a. In certain embodiments, R^6a is R^7b. In certain embodiments, R^6b is R^7b. In certain embodiments, R^6a is - CH2CH2C(=O)OR^7a. In certain embodiments, R^6b is -CH2CH2C(=O)OR^7a. In certain embodiments, R^7a is optionally substituted C₆-C₂₈ alkyl. In certain embodiments, R^7a is optionally substituted C6-C28 alkenyl. In certain embodiments, R^7a is optionally substituted C₆-C₂₈ heteroalkyl. In certain embodiments, R^7b is optionally substituted C6-C28 alkyl. In certain embodiments, R^7b is optionally substituted C₆-C₂₈ alkenyl. In certain embodiments, R^7b is optionally substituted C6-C28 heteroalkyl. In certain embodiments, R^6a is -CH₂CH(OH)(optionally substituted C₆-C₂₈ alkyl). In certain embodiments, R^6a is CH2CH(OH)(optionally substituted C6-C28 alkenyl). In certain embodiments, R^6a is -CH₂CH(OH)(optionally substituted C₆-C₂₈ heteroalkyl). In certain embodiments, R^6a is -CH2CH2C(=O)O(optionally substituted C6-C28 alkyl). In certain embodiments, R^6a is -CH2CH2C(=O)O(optionally substituted C6-C28 alkenyl). In certain - 53 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) embodiments, R^6a is -CH2CH2C(=O)O(optionally substituted C6-C28 heteroalkyl). In certain embodiments, R^6b is -CH₂CH(OH)(optionally substituted C₆-C₂₈ alkyl). In certain embodiments, R^6b is CH2CH(OH)(optionally substituted C6-C28 alkenyl). In certain embodiments, R^6b is -CH₂CH(OH)(optionally substituted C₆-C₂₈ heteroalkyl). In certain embodiments, R^6b is -CH2CH2C(=O)O(optionally substituted C6-C28 alkyl). In certain embodiments, R^6b is -CH₂CH₂C(=O)O(optionally substituted C₆-C₂₈ alkenyl). In certain embodiments, R^6b is -CH2CH2C(=O)O(optionally substituted C6-C28 heteroalkyl). In certain embodiments, R^6a is -CH₂CH(OH)(CH₂)₉CH₃. In certain embodiments, R^6a is -CH2CH2C(=O)O(CH2)11CH3. In certain embodiments, R^6b is -CH₂CH(OH)(CH₂)₉CH₃. In certain embodiments, R^6b is -CH2CH2C(=O)O(CH2)11CH3. In certain embodiments, the compound is: . In . of a nucleic acid, small molecule, protein, therapeutic agent, antibody, and any combinations thereof. In certain embodiments, the cargo is a nucleic acid. In certain embodiments, the nucleic acid is DNA or RNA. In certain embodiments, the nucleic acid is selected from the group consisting of mRNA, cDNA, pDNA, microRNA, 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 cargo is mRNA. In certain embodiments, the mRNA encodes SARS-CoV-2, an immunogenic fragment thereof (e.g., spike protein), or a modified derivative thereof. - 54 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) In certain embodiments, the infection, disease, or disorder is a SARS-CoV-2 infection. In certain embodiments, an innate immune response is promoted in the subject. In certain embodiments, the subject is a mammal. In certain embodiments, the mammal is a human. Cargo In one aspect, the present disclosure relates to LNPs comprising at least one cargo molecule at least partially encapsulated therein. In certain embodiments, the at least one cargo is fully encapsulated therein. Small molecule therapeutic agents In various embodiments, the agent is a therapeutic agent. In various embodiments, the therapeutic agent is a small molecule. When the therapeutic agent is a small molecule, a small molecule may be obtained using standard methods known to the skilled artisan. Such methods include chemical organic synthesis or biological means. Biological means include purification from a biological source, recombinant synthesis and in vitro translation systems, using methods well known in the art. In certain embodiments, a small molecule therapeutic agents comprises an organic molecule, inorganic molecule, biomolecule, synthetic molecule, and the like. Combinatorial libraries of molecularly diverse chemical compounds potentially useful in treating a variety of diseases and conditions are well known in the art, as are method of making the libraries. The method may use a variety of techniques well-known to the skilled artisan including solid phase synthesis, solution methods, parallel synthesis of single compounds, synthesis of chemical mixtures, rigid core structures, flexible linear sequences, deconvolution strategies, tagging techniques, and generating unbiased molecular landscapes for lead discovery vs. biased structures for lead development. In some embodiments of the disclosure, the therapeutic agent is synthesized and/or identified using combinatorial techniques. In a general method for small library synthesis, an activated core molecule is condensed with a number of building blocks, resulting in a combinatorial library of covalently linked, core-building block ensembles. The shape and rigidity of the core determines the orientation of the building blocks in shape space. The libraries can be biased by changing the core, linkage, or building blocks to target a characterized biological structure - 55 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) ("focused libraries") or synthesized with less structural bias using flexible cores. In some embodiments of the disclosure, the therapeutic agent is synthesized via small library synthesis. The small molecule and small molecule compounds described herein may be present as salts even if salts are not depicted, and it is understood that the disclosure embraces all salts and solvates of the therapeutic agents depicted here, as well as the non-salt and non- solvate form of the therapeutic agents, as is well understood by the skilled artisan. In some embodiments, the salts of the therapeutic agents of the disclosure are pharmaceutically acceptable salts. Where tautomeric forms may be present for any of the therapeutic agents described herein, each and every tautomeric form is intended to be included in the present disclosure, even though only one or some of the tautomeric forms may be explicitly depicted. For example, when a 2-hydroxypyridyl moiety is depicted, the corresponding 2-pyridone tautomer is also intended. The disclosure also includes any or all of the stereochemical forms, including any enantiomeric or diastereomeric forms of the therapeutic agents described. The recitation of the structure or name herein is intended to embrace all possible stereoisomers of therapeutic agents depicted. All forms of the therapeutic agents are also embraced by the disclosure, such as crystalline or non-crystalline forms of the therapeutic agent. Compositions comprising a therapeutic agents of the disclosure are also intended, such as a composition of substantially pure therapeutic agent, including a specific stereochemical form thereof, or a composition comprising mixtures of therapeutic agents of the disclosure in any ratio, including two or more stereochemical forms, such as in a racemic or non-racemic mixture. The disclosure also includes any or all active analog or derivative, such as a prodrug, of any therapeutic agent described herein. In certain embodiments, the therapeutic agent is a prodrug. In certain embodiments, the small molecules described herein are candidates for derivatization. As such, in certain instances, the analogs of the small molecules described herein that have modulated potency, selectivity, and solubility are included herein and provide useful leads for drug discovery and drug development. Thus, in certain instances, during optimization new analogs are designed considering issues of drug delivery, metabolism, novelty, and safety. In some instances, small molecule therapeutic agents described herein are derivatives or analogs of known therapeutic agents, as is well known in the art of combinatorial and medicinal chemistry. The analogs or derivatives can be prepared by adding and/or - 56 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) substituting functional groups at various locations. As such, the small molecules described herein can be converted into derivatives/analogs using well known chemical synthesis procedures. For example, all of the hydrogen atoms or substituents can be selectively modified to generate new analogs. Also, the linking atoms or groups can be modified into longer or shorter linkers with carbon backbones or hetero atoms. Also, the ring groups can be changed so as to have a different number of atoms in the ring and/or to include hetero atoms. Moreover, aromatics can be converted to cyclic rings, and vice versa. For example, the rings may be from 5-7 atoms, and may be carbocyclic or heterocyclic. As used herein, the term "analog," "analogue," or "derivative" is meant to refer to a chemical compound or molecule made from a parent compound or molecule by one or more chemical reactions. As such, an analog can be a structure having a structure similar to that of the small molecule therapeutic agents described herein or can be based on a scaffold of a small molecule therapeutic agents described herein, but differing from it in respect to certain components or structural makeup, which may have a similar or opposite action metabolically. An analog or derivative of any of a small molecule inhibitor in accordance with the present disclosure can be used to treat a disease or disorder. In certain embodiments, the small molecule therapeutic agents described herein can independently be derivatized, or analogs prepared therefrom, by modifying hydrogen groups independently from each other into other substituents. That is, each atom on each molecule can be independently modified with respect to the other atoms on the same molecule. Any traditional modification for producing a derivative/analog can be used. For example, the atoms and substituents can be independently comprised of hydrogen, an alkyl, aliphatic, straight chain aliphatic, aliphatic having a chain hetero atom, branched aliphatic, substituted aliphatic, cyclic aliphatic, heterocyclic aliphatic having one or more hetero atoms, aromatic, heteroaromatic, polyaromatic, polyamino acids, peptides, polypeptides, combinations thereof, halogens, halo-substituted aliphatics, and the like. Additionally, any ring group on a compound can be derivatized to increase and/or decrease ring size as well as change the backbone atoms to carbon atoms or hetero atoms. Nucleic acid therapeutic agents In certain embodiments, the composition of the disclosure comprises an in vitro transcribed (IVT) RNA molecule. For example, in certain embodiments, the composition of the disclosure comprises an IVT RNA molecule which encodes an agent. In certain embodiments, the IVT RNA molecule of the present composition is a nucleoside-modified - 57 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) mRNA molecule. In certain embodiments, the agent is for targeting an immune cell to a pathogen or a tumor cell of interest. In certain embodiments, the IVT RNA molecule encodes a chimeric antigen receptor (CAR). In some embodiments, the CAR is specific for binding to one or more antigens. In some embodiments, the antigen comprises at least one viral antigen, a bacterial antigen, a fungal antigen, a parasitic antigen, an influenza antigen, a tumor-associated antigen, a tumor- specific antigen, or any combination thereof. However, the present disclosure is not limited to any particular agent or combination of agents. In certain embodiments, the composition comprises an adjuvant. In certain embodiments, the composition comprises a nucleic acid molecule encoding an adjuvant. In certain embodiments, the composition comprises a nucleoside-modified RNA encoding an adjuvant. In certain embodiments, the composition comprises at least one RNA molecule encoding a combination of at least two agents. In certain embodiments, the composition comprises a combination of two or more RNA molecules encoding a combination of two or more agents. In certain embodiments, the present disclosure provides a method for inducing an immune response in a subject. For example, the method can be used to provide immunity in the subject against a virus, bacteria, fungus, parasite, cancer, or the like. In some embodiments, the method comprises administering to the subject a composition comprising one or more LNP molecule formulated for in vivo targeting of an immune cell comprising one or more RNA encoding at least one antigen, an adjuvant, or a combination thereof. In certain embodiments, the present disclosure provides a method for gene editing of an immune cell of a subject. For example, the method can be used to provide one or more component of a gene editing system (e.g., a component of a CRISPR system) to an immune cell of a subject. In some embodiments, the method comprises administering to the subject a composition comprising one or more ionizable LNP molecule formulated for targeted T cell delivery comprising one or more nucleoside-modified RNA molecule for gene editing. In certain embodiments, the method comprises administration of the composition to a subject. In certain embodiments, the method comprises administering a plurality of doses to the subject. In some embodiments, the method comprises administering a single dose of the composition, where the single dose is effective in delivery of the target therapeutic agent. In other related aspects, the therapeutic agent is an isolated nucleic acid. In certain embodiments, the isolated nucleic acid molecule is one of a DNA molecule or an RNA - 58 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) molecule. In certain embodiments, the isolated nucleic acid molecule is a cDNA, mRNA, siRNA, shRNA or miRNA molecule. In certain embodiments, the isolated nucleic acid molecule encodes a therapeutic peptide such a thrombomodulin, endothelial protein C receptor (EPCR), anti-thrombotic proteins including plasminogen activators and their mutants, antioxidant proteins including catalase, superoxide dismutase (SOD) and iron- sequestering proteins. In some embodiments, the therapeutic agent is an siRNA, miRNA, shRNA, or an antisense molecule, which inhibits a targeted nucleic acid including those encoding proteins that are involved in aggravation of the pathological processes. In certain embodiments, the nucleic acid comprises a promoter/regulatory sequence such that the nucleic acid is capable of directing expression of the nucleic acid. Thus, the disclosure encompasses expression vectors and methods for the introduction of exogenous nucleic acid into cells with concomitant expression of the exogenous nucleic acid in the cells such as those described, for example, in Sambrook et al. (2012, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, New York), and in Ausubel et al. (1997, Current Protocols in Molecular Biology, John Wiley & Sons, New York) and as described elsewhere herein. In certain embodiments, siRNA is used to decrease the level of a targeted protein. RNA interference (RNAi) is a phenomenon in which the introduction of double-stranded RNA (dsRNA) into a diverse range of organisms and cell types causes degradation of the complementary mRNA. In the cell, long dsRNAs are cleaved into short 21-25 nucleotide small interfering RNAs, or siRNAs, by a ribonuclease known as Dicer. The siRNAs subsequently assemble with protein components into an RNA-induced silencing complex (RISC), unwinding in the process. Activated RISC then binds to complementary transcript by base pairing interactions between the siRNA antisense strand and the mRNA. The bound mRNA is cleaved and sequence specific degradation of mRNA results in gene silencing. See, for example, U.S. Patent No.6,506,559; Fire et al., 1998, Nature 391(19):306-311; Timmons et al., 1998, Nature 395:854; Montgomery et al., 1998, TIG 14 (7):255-258; David R. Engelke, Ed., RNA Interference (RNAi) Nuts & Bolts of RNAi Technology, DNA Press, Eagleville, PA (2003); and Gregory J. Hannon, Ed., RNAi A Guide to Gene Silencing, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (2003). Soutschek et al. (2004, Nature 432:173-178) describe a chemical modification to siRNAs that aids in intravenous systemic delivery. Optimizing siRNAs involves consideration of overall G/C content, C/T content at the termini, Tm and the nucleotide content of the 3' overhang. See, for instance, Schwartz et al., 2003, Cell, 115:199-208 and Khvorova et al., 2003, Cell 115:209-216. - 59 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) Therefore, the present disclosure also includes methods of decreasing levels of PTPN22 using RNAi technology. In one aspect, the disclosure includes a vector comprising an siRNA or an antisense polynucleotide. Preferably, the siRNA or antisense polynucleotide is capable of inhibiting the expression of a target polypeptide. The incorporation of a desired polynucleotide into a vector and the choice of vectors are well-known in the art as described in, for example, Sambrook et al. (2012), and in Ausubel et al. (1997), and elsewhere herein. In certain embodiments, the expression vectors described herein encode a short hairpin RNA (shRNA) therapeutic agents. shRNA molecules are well known in the art and are directed against the mRNA of a target, thereby decreasing the expression of the target. In certain embodiments, the encoded shRNA is expressed by a cell, and is then processed into siRNA. For example, in certain instances, the cell possesses native enzymes (e.g., dicer) that cleave the shRNA to form siRNA. In order to assess the expression of the siRNA, shRNA, or antisense polynucleotide, the expression vector to be introduced into a cell can also contain either a selectable marker gene or a reporter gene or both to facilitate identification of expressing cells from the population of cells sought to be transfected or infected using a the delivery vehicle of the disclosure. In other embodiments, the selectable marker may be carried on a separate piece of DNA and also be contained within the delivery vehicle. Both selectable markers and reporter genes may be flanked with appropriate regulatory sequences to enable expression in the host cells. Useful selectable markers are known in the art and include, for example, antibiotic- resistance genes, such as neomycin resistance and the like. Therefore, in one aspect, the delivery vehicle may contain a vector, comprising the nucleotide sequence or the construct to be delivered. The choice of the vector will depend on the host cell in which it is to be subsequently introduced. In a particular embodiment, the vector of the disclosure is an expression vector. Suitable host cells include a wide variety of prokaryotic and eukaryotic host cells. In specific embodiments, the expression vector is selected from the group consisting of a viral vector, a bacterial vector and a mammalian cell vector. Prokaryote- and/or eukaryote-vector based systems can be employed for use with the present disclosure to produce polynucleotides, or their cognate polypeptides. Many such systems are commercially and widely available. By way of illustration, the vector in which the nucleic acid sequence is introduced can be a plasmid, which is or is not integrated in the genome of a host cell when it is introduced in the cell. Illustrative, non-limiting examples of vectors in which the nucleotide sequence of - 60 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) the disclosure or the gene construct of the disclosure can be inserted include a tet-on inducible vector for expression in eukaryote cells. The vector may be obtained by conventional methods known by persons skilled in the art (Sambrook et al., 2012). In a particular embodiment, the vector is a vector useful for transforming animal cells. In certain embodiments, the recombinant expression vectors may also contain nucleic acid molecules, which encode a peptide or peptidomimetic. A promoter may be one naturally associated with a gene or polynucleotide sequence, as may be obtained by isolating the 5' non-coding sequences located upstream of the coding segment and/or exon. Such a promoter can be referred to as "endogenous." Similarly, an enhancer may be one naturally associated with a polynucleotide sequence, located either downstream or upstream of that sequence. Alternatively, certain advantages will be gained by positioning the coding polynucleotide segment under the control of a recombinant or heterologous promoter, which refers to a promoter that is not normally associated with a polynucleotide sequence in its natural environment. A recombinant or heterologous enhancer refers also to an enhancer not normally associated with a polynucleotide sequence in its natural environment. Such promoters or enhancers may include promoters or enhancers of other genes, and promoters or enhancers isolated from any other prokaryotic, viral, or eukaryotic cell, and promoters or enhancers not "naturally occurring," i.e., containing different elements of different transcriptional regulatory regions, and/or mutations that alter expression. In addition to producing nucleic acid sequences of promoters and enhancers synthetically, sequences may be produced using recombinant cloning and/or nucleic acid amplification technology, including PCR™, in connection with the compositions disclosed herein (U.S. Patent 4,683,202, U.S. Patent 5,928,906). Furthermore, it is contemplated the control sequences that direct transcription and/or expression of sequences within non-nuclear organelles such as mitochondria, chloroplasts, and the like, can be employed as well. Naturally, it will be important to employ a promoter and/or enhancer that effectively directs the expression of the DNA segment in the cell type, organelle, and organism chosen for expression. Those of skill in the art of molecular biology generally know how to use promoters, enhancers, and cell type combinations for protein expression, for example, see Sambrook et al. (2012). The promoters employed may be constitutive, tissue-specific, inducible, and/or useful under the appropriate conditions to direct high level expression of the introduced DNA segment, such as is advantageous in the large-scale production of recombinant proteins and/or peptides. The promoter may be heterologous or endogenous. - 61 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) The recombinant expression vectors may also contain a selectable marker gene, which facilitates the selection of host cells. Suitable selectable marker genes are genes encoding proteins such as G418 and hygromycin, which confer resistance to certain drugs, β- galactosidase, chloramphenicol acetyltransferase, firefly luciferase, or an immunoglobulin or portion thereof such as the Fc portion of an immunoglobulin preferably IgG. The selectable markers may be introduced on a separate vector from the nucleic acid of interest. Following the generation of the siRNA polynucleotide, a skilled artisan will understand that the siRNA polynucleotide will have certain characteristics that can be modified to improve the siRNA as a therapeutic compound. Therefore, the siRNA polynucleotide may be further designed to resist degradation by modifying it to include phosphorothioate, or other linkages, methylphosphonate, sulfone, sulfate, ketyl, phosphorodithioate, phosphoramidate, phosphate esters, and the like (see, e.g., Agrawal et al., 1987, Tetrahedron Lett.28:3539-3542; Stec et al., 1985 Tetrahedron Lett.26:2191-2194; Moody et al., 1989 Nucleic Acids Res.12:4769-4782; Eckstein, 1989 Trends Biol. Sci. 14:97-100; Stein, In: Oligodeoxynucleotides. Antisense Inhibitors of Gene Expression, Cohen, ed., Macmillan Press, London, pp.97-117 (1989)). Any polynucleotide may be further modified to increase its stability in vivo. Possible modifications include, but are not limited to, the addition of flanking sequences at the 5' and/or 3' ends; the use of phosphorothioate or 2' O-methyl rather than phosphodiester linkages in the backbone; and/or the inclusion of nontraditional bases such as inosine, queuosine, and wybutosine and the like, as well as acetyl- methyl-, thio- and other modified forms of adenine, cytidine, guanine, thymine, and uridine. In certain embodiments of the disclosure, an antisense nucleic acid sequence, which is expressed by a plasmid vector is used as a therapeutic agent to inhibit the expression of a target protein. The antisense expressing vector is used to transfect a mammalian cell or the mammal itself, thereby causing reduced endogenous expression of the target protein. Antisense molecules and their use for inhibiting gene expression are well known in the art (see, e.g., Cohen, 1989, In: Oligodeoxyribonucleotides, Antisense Inhibitors of Gene Expression, CRC Press). Antisense nucleic acids are DNA or RNA molecules that are complementary, as that term is defined elsewhere herein, to at least a portion of a specific mRNA molecule (Weintraub, 1990, Scientific American 262:40). In the cell, antisense nucleic acids hybridize to the corresponding mRNA, forming a double-stranded molecule thereby inhibiting the translation of genes. The use of antisense methods to inhibit the translation of genes is known in the art, - 62 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) and is described, for example, in Marcus-Sakura (1988, Anal. Biochem.172:289). Such antisense molecules may be provided to the cell via genetic expression using DNA encoding the antisense molecule as taught by Inoue, 1993, U.S. Patent No.5,190,931. Alternatively, antisense molecules of the disclosure may be made synthetically and then provided to the cell. Antisense oligomers of between about 10 to about 30, and more preferably about 15 nucleotides, are preferred, since they are easily synthesized and introduced into a target cell. Synthetic antisense molecules contemplated by the disclosure include oligonucleotide derivatives known in the art which have improved biological activity compared to unmodified oligonucleotides (see U.S. Patent No.5,023,243). In certain embodiments of the disclosure, a ribozyme is used as a therapeutic agent to inhibit expression of a target protein. Ribozymes useful for inhibiting the expression of a target molecule may be designed by incorporating target sequences into the basic ribozyme structure, which are complementary, for example, to the mRNA sequence encoding the target molecule. Ribozymes targeting the target molecule, may be synthesized using commercially available reagents (Applied Biosystems, Inc., Foster City, CA) or they may be genetically expressed from DNA encoding them. In certain embodiments, the therapeutic agent may comprise one or more components of a CRISPR-Cas system, where a guide RNA (gRNA) targeted to a gene encoding a target molecule, and a CRISPR-associated (Cas) peptide form a complex to induce mutations within the targeted gene. In certain embodiments, the therapeutic agent comprises a gRNA or a nucleic acid molecule encoding a gRNA. In certain embodiments, the therapeutic agent comprises a Cas peptide or a nucleic acid molecule encoding a Cas peptide. In certain embodiments, the agent comprises a miRNA or a mimic of a miRNA. In certain embodiments, the agent comprises a nucleic acid molecule that encodes a miRNA or mimic of a miRNA. MiRNAs are small non-coding RNA molecules that are capable of causing post- transcriptional silencing of specific genes in cells by the inhibition of translation or through degradation of the targeted mRNA. A miRNA can be completely complementary or can have a region of noncomplementarity with a target nucleic acid, consequently resulting in a "bulge" at the region of non-complementarity. A miRNA can inhibit gene expression by repressing translation, such as when the miRNA is not completely complementary to the target nucleic acid, or by causing target RNA degradation, which is believed to occur only when the miRNA binds its target with perfect complementarity. The disclosure also can include double-stranded precursors of miRNA. A miRNA or pri-miRNA can be 18- 100 - 63 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) nucleotides in length, or from 18-80 nucleotides in length. Mature miRNAs can have a length of 19-30 nucleotides, or 21-25 nucleotides, particularly 21, 22, 23, 24, or 25 nucleotides. MiRNA precursors typically have a length of about 70-100 nucleotides and have a hairpin conformation. miRNAs are generated in vivo from pre- miRNAs by the enzymes Dicer and Drosha, which specifically process long pre-miRNA into functional miRNA. The hairpin or mature microRNAs, or pri-microRNA agents featured in the disclosure can be synthesized in vivo by a cell-based system or in vitro by chemical synthesis. In various embodiments, the agent comprises an oligonucleotide that comprises the nucleotide sequence of a disease-associated miRNA. In certain embodiments, the oligonucleotide comprises the nucleotide sequence of a disease-associated miRNA in a pre - microRNA, mature or hairpin form. In other embodiments, a combination of oligonucleotides comprising a sequence of one or more disease-associated miRNAs, any pre -miRNA, any fragment, or any combination thereof is envisioned. MiRNAs can be synthesized to include a modification that imparts a desired characteristic. For example, the modification can improve stability, hybridization thermodynamics with a target nucleic acid, targeting to a particular tissue or cell -type, or cell permeability, e.g., by an endocytosis-dependent or -independent mechanism. Modifications can also increase sequence specificity, and consequently decrease off- site targeting. Methods of synthesis and chemical modifications are described in greater detail below. If desired, miRNA molecules may be modified to stabilize the miRNAs against degradation, to enhance half-life, or to otherwise improve efficacy. Desirable modifications are described, for example, in U.S. Patent Publication Nos.20070213292, 20060287260, 20060035254.20060008822. and 2005028824, each of which is hereby incorporated by reference in its entirety. For increased nuclease resistance and/or binding affinity to the target, the single- stranded oligonucleotide agents featured in the disclosure can include 2'-O- methyl, 2'-fluorine, 2'-O-methoxyethyl, 2'-O-aminopropyl, 2'-amino, and/or phosphorothioate linkages. Inclusion of locked nucleic acids (LNA), ethylene nucleic acids (ENA), e.g., 2'-4'- ethylene- bridged nucleic acids, and certain nucleotide modifications can also increase binding affinity to the target. The inclusion of pyranose sugars in the oligonucleotide backbone can also decrease endonucleolytic cleavage. An oligonucleotide can be further modified by including a 3' cationic group, or by inverting the nucleoside at the 3'-terminus with a 3 -3' linkage. In another alternative, the 3 '-terminus can be blocked with an aminoalkyl group. Other 3' conjugates can inhibit 3'-5' exonucleolytic cleavage. While not being bound by theory, a 3' may inhibit exonucleolytic cleavage by sterically blocking the - 64 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) exonuclease from binding to the 3' end of the oligonucleotide. Even small alkyl chains, aryl groups, or heterocyclic conjugates or modified sugars (D-ribose, deoxyribose, glucose, and so forth) can block 3'-5'-exonucleases. In certain embodiments, the miRNA includes a 2'-modified oligonucleotide containing oligodeoxynucleotide gaps with some or all internucleotide linkages modified to phosphorothioates for nuclease resistance. The presence of methylphosphonate modifications increases the affinity of the oligonucleotide for its target RNA and thus reduces the IC5Q. This modification also increases the nuclease resistance of the modified oligonucleotide. It is understood that the methods and reagents of the present disclosure may be used in conjunction with any technologies that may be developed to enhance the stability or efficacy of an inhibitory nucleic acid molecule. miRNA molecules include nucleotide oligomers containing modified backbones or non-natural internucleoside linkages. Oligomers having modified backbones include those that retain a phosphorus atom in the backbone and those that do not have a phosphorus atom in the backbone. For the purposes of this disclosure, modified oligonucleotides that do not have a phosphorus atom in their internucleoside backbone are also considered to be nucleotide oligomers. Nucleotide oligomers that have modified oligonucleotide backbones include, for example, phosphorothioates, chiral phosphorothioates, phosphorodithioates, phosphotriesters, aminoalkyl-phosphotriesters, methyl and other alkyl phosphonates including 3'-alkylene phosphonates and chiral phosphonates, phosphinates, phosphoramidates, thionophosphoramidates, thionoalkylphosphonates, thionoalkylphosphotriest- ers, and boranophosphates. Various salts, mixed salts and free acid forms are also included. A miRNA described herein, which may be in the mature or hairpin form, may be provided as a naked oligonucleotide. In some cases, it may be desirable to utilize a formulation that aids in the delivery of a miRNA or other nucleotide oligomer to cells (see, e.g., U.S. Patent Nos.5,656,611, 5,753,613, 5,785,992, 6,120,798, 6,221,959, 6,346,613, and 6,353,055, each of which is hereby incorporated by reference). In some examples, the miRNA composition is at least partially crystalline, uniformly crystalline, and/or anhydrous (e.g., less than 80, 50, 30, 20, or 10% water). In another example, the miRNA composition is in an aqueous phase, e.g., in a solution that includes water. The aqueous phase or the crystalline compositions can be incorporated into a delivery vehicle, e.g., a liposome (particularly for the aqueous phase), or a particle (e.g., a microparticle as can be appropriate for a crystalline composition). Generally, the miRNA - 65 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) composition is formulated in a manner that is compatible with the intended method of administration. A miRNA composition can be formulated in combination with another agent, e.g., another therapeutic agent or an agent that stabilizes an oligonucleotide agent, e.g., a protein that complexes with the oligonucleotide agent. Still other agents include chelators, e.g., EDTA (e.g., to remove divalent cations such as Mg), salts, and RNAse inhibitors (e.g., a broad specificity RNAse inhibitor). In certain embodiments, the miRNA composition includes another miRNA, e.g., a second miRNA composition (e.g., a microRNA that is distinct from the first). Still other preparations can include at least three, five, ten, twenty, fifty, or a hundred or more different oligonucleotide species. In certain embodiments, the composition comprises an oligonucleotide composition that mimics the activity of a miRNA. In certain embodiments, the composition comprises oligonucleotides having nucleobase identity to the nucleobase sequence of a miRNA, and are thus designed to mimic the activity of the miRNA. In certain embodiments, the oligonucleotide composition that mimics miRNA activity comprises a double-stranded RNA molecule which mimics the mature miRNA hairpins or processed miRNA duplexes. In certain embodiments, the oligonucleotide shares identity with endogenous miRNA or miRNA precursor nucleobase sequences. An oligonucleotide selected for inclusion in a composition of the present disclosure may be one of a number of lengths. Such an oligonucleotide can be from 7 to 100 linked nucleosides in length. For example, an oligonucleotide sharing nucleobase identity with a miRNA may be from 7 to 30 linked nucleosides in length. An oligonucleotide sharing identity with a miRNA precursor may be up to 100 linked nucleosides in length. In certain embodiments, an oligonucleotide comprises 7 to 30 linked nucleosides. In certain embodiments, an oligonucleotide comprises 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 28, 29, or 30 linked nucleotides. In certain embodiments, an oligonucleotide comprises 19 to 23 linked nucleosides. In certain embodiments, an oligonucleotide is from 40 up to 50, 60, 70, 80, 90, or 100 linked nucleosides in length. In certain embodiments, an oligonucleotide has a sequence that has a certain identity to a miRNA or a precursor thereof. Nucleobase sequences of mature miRNAs and their corresponding stem-loop sequences described herein are the sequences found in miRBase, an online searchable database of miRNA sequences and annotation. Entries in the miRBase Sequence database represent a predicted hairpin portion of a miRNA transcript (the stem- loop), with information on the location and sequence of the mature miRNA sequence. The miRNA stem-loop sequences in the database are not strictly precursor miRNAs (pre- - 66 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) miRNAs), and may in some instances include the pre-miRNA and some flanking sequence from the presumed primary transcript. The miRNA nucleobase sequences described herein encompass any version of the miRNA, including the sequences described in Release 10.0 of the miRBase sequence database and sequences described in any earlier Release of the miRBase sequence database. A sequence database release may result in the re-naming of certain miRNAs. A sequence database release may result in a variation of a mature miRNA sequence. The compositions of the present disclosure encompass oligomeric compound comprising oligonucleotides having a certain identity to any nucleobase sequence version of a miRNAs described herein. In certain embodiments, an oligonucleotide has a nucleobase sequence at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98% or 99% identical to the miRNA over a region of 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleobases. Accordingly, in certain embodiments the nucleobase sequence of an oligonucleotide may have one or more non-identical nucleobases with respect to the miRNA. In certain embodiments, the composition comprises a nucleic acid molecule encoding a miRNA, precursor, mimic, or fragment thereof. For example, the composition may comprise a viral vector, plasmid, cosmid, or other expression vector suitable for expressing the miRNA, precursor, mimic, or fragment thereof in a desired mammalian cell or tissue. Combinations In certain embodiments, the composition of the present disclosure comprises a combination of agents described herein. In certain embodiments, a composition comprising a combination of agents described herein has an additive effect, wherein the overall effect of the combination is approximately equal to the sum of the effects of each individual agent. In other embodiments, a composition comprising a combination of agents described herein has a synergistic effect, wherein the overall effect of the combination is greater than the sum of the effects of each individual agent. A composition comprising a combination of agents comprises individual agents in any suitable ratio. For example, In certain embodiments, the composition comprises a 1:1 ratio of two individual agents. However, the combination is not limited to any particular ratio. Rather any ratio that is shown to be effective is encompassed. Pharmaceutical Compositions The formulations of the pharmaceutical compositions described herein may be - 67 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) prepared by any method known or hereafter developed in the art of pharmacology. 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- or multi-dose unit. Although the description of pharmaceutical compositions provided herein are principally directed to pharmaceutical compositions which are 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 disclosure is contemplated include, but are not limited to, humans and other primates, mammals including commercially relevant mammals such as non-human primates, cattle, pigs, horses, sheep, cats, and dogs. Pharmaceutical compositions that are useful in the methods of the disclosure may be prepared, packaged, or sold in formulations suitable for ophthalmic, oral, rectal, vaginal, parenteral, topical, pulmonary, intranasal, buccal, intravenous, intracerebroventricular, intradermal, intramuscular, or another route of administration. Other contemplated formulations include projected nanoparticles, liposomal preparations, resealed erythrocytes containing the active ingredient, and immunogenic-based formulations. A pharmaceutical composition of the disclosure may be prepared, packaged, or sold in bulk, as a single unit dose, or as a plurality of single unit doses. As used herein, a "unit dose" is 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 which 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 relative amounts of the active ingredient, the pharmaceutically acceptable carrier, and any additional ingredients in a pharmaceutical composition of the disclosure 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. In addition to the active ingredient, a pharmaceutical composition of the disclosure may further comprise one or more additional pharmaceutically active agents. - 68 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) Controlled- or sustained-release formulations of a pharmaceutical composition of the disclosure may be made using conventional technology. 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, intraocular, intravitreal, subcutaneous, intraperitoneal, intramuscular, intradermal, intrasternal injection, intratumoral, intravenous, intracerebroventricular 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 multi-dose containers containing a preservative. Formulations for parenteral administration include, but are not limited to, suspensions, solutions, emulsions in oily or aqueous vehicles, pastes, and implantable sustained-release or biodegradable formulations. Such formulations may further comprise one or more additional ingredients including, but not limited to, suspending, stabilizing, or dispersing agents. In certain embodiments of a formulation for parenteral administration, the active ingredient is provided in dry (i.e. powder or granular) form for reconstitution with a suitable vehicle (e.g. sterile pyrogen-free water) prior to parenteral administration of the reconstituted composition. The pharmaceutical compositions may be prepared, packaged, or sold in the form of a sterile injectable aqueous or oily suspension or solution. This suspension or solution may be formulated according to the known art, and may comprise, in addition to the active ingredient, additional ingredients such as the dispersing agents, wetting agents, or suspending agents described herein. Such sterile injectable formulations may be prepared using a non-toxic parenterally-acceptable diluent or solvent, such as water or 1,3-butane diol, 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 - 69 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) comprise the active ingredient in microcrystalline form, in a liposomal preparation, or as a component of a biodegradable polymer systems. 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. A pharmaceutical composition of the disclosure may be prepared, packaged, or sold in a formulation suitable for pulmonary administration via the buccal cavity. Such a formulation may comprise dry particles which comprise the active ingredient and which have a diameter in the range from about 0.5 to about 7 micrometers, and preferably from about 1 to about 6 micrometers. Such compositions are conveniently in the form of dry powders for administration using a device comprising a dry powder reservoir to which a stream of propellant may be directed to disperse the powder or using a self-propelling solvent/powder-dispensing container such as a device comprising the active ingredient dissolved or suspended in a low-boiling propellant in a sealed container. Preferably, such powders comprise particles wherein at least 98% of the particles by weight have a diameter greater than 0.5 micrometers and at least 95% of the particles by number have a diameter less than 7 micrometers. More preferably, at least 95% of the particles by weight have a diameter greater than 1 micrometer and at least 90% of the particles by number have a diameter less than 6 micrometers. Dry powder compositions preferably include a solid fine powder diluent such as sugar and are conveniently provided in a unit dose form. Low boiling propellants generally include liquid propellants having a boiling point of below 65°F at atmospheric pressure. Generally the propellant may constitute 50 to 99.9% (w/w) of the composition, and the active ingredient may constitute 0.1 to 20% (w/w) of the composition. The propellant may further comprise additional ingredients such as a liquid non-ionic or solid anionic surfactant or a solid diluent (preferably having a particle size of the same order as particles comprising the active ingredient). 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 multi-dose containers containing a preservative. Formulations for parenteral administration include, but are not limited to, suspensions, solutions, emulsions in oily or aqueous vehicles, pastes, and implantable sustained-release or biodegradable formulations. Such formulations - 70 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) 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-butane diol, 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 that are useful include those that comprise the active ingredient in microcrystalline form, 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. As used herein, "additional ingredients" include, but are not limited to, one or more of the following: excipients; surface active agents; dispersing agents; inert diluents; granulating and disintegrating agents; binding agents; lubricating agents; sweetening agents; flavoring agents; coloring agents; preservatives; physiologically degradable compositions such as gelatin; aqueous vehicles and solvents; oily vehicles and solvents; suspending agents; dispersing or wetting agents; emulsifying agents, demulcents; buffers; salts; thickening agents; fillers; emulsifying agents; antioxidants; antibiotics; antifungal agents; stabilizing agents; and pharmaceutically acceptable polymeric or hydrophobic materials. Other "additional ingredients" which may be included in the pharmaceutical compositions of the disclosure are known in the art and described, for example in Remington's Pharmaceutical Sciences (1985, Genaro, ed., Mack Publishing Co., Easton, PA), which is incorporated herein by reference. Administration/Dosing The regimen of administration may affect what constitutes an effective amount. The - 71 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) therapeutic formulations may be administered to the patient either prior to or after the onset of a disease or disorder. Further, several divided dosages, as well as staggered dosages may be administered daily or sequentially, or the dose may be continuously infused, or may be a bolus injection. Further, the dosages of the therapeutic formulations may be proportionally increased or decreased as indicated by the exigencies of the therapeutic or prophylactic situation. Administration of the compositions of the present disclosure to a patient, such as a mammal, such as a human, may be carried out using known procedures, at dosages and for periods of time effective to treat a disease or disorder contemplated herein. An effective amount of therapeutic (i.e., composition) necessary to achieve a therapeutic effect may vary according to factors such as the activity of the particular therapeutic employed; the time of administration; the rate of excretion of the composition; the duration of the treatment; other drugs, compounds or materials used in combination with the composition; the state of the disease or disorder, age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well-known in the medical arts. Dosage regimens may be adjusted to provide the optimum therapeutic response. For example, several divided doses may be administered daily or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation. A non-limiting example of an effective dose range for a therapeutic composition of the disclosure is from about 0.01 mg/kg to 100 mg/kg of body weight/per day of active agent (i.e., nucleic acid). One of ordinary skill in the art would be able to study the relevant factors and make the determination regarding the effective amount of the therapeutic composition without undue experimentation. The composition may be administered to an animal as frequently as several times daily, or it may be administered less frequently, such as once a day, once a week, once every two weeks, once a month, or even less frequently, such as once every several months or even once a year or less. It is understood that the amount of composition dosed per day may be 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 - 72 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) this disclosure may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient. A medical doctor, e.g., physician or veterinarian, having ordinary skill in the art may readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the compounds of the disclosure employed in the pharmaceutical composition at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. In particular embodiments, it is especially advantageous to formulate the compound in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the patients to be treated; each unit containing a predetermined quantity of therapeutic composition to produce the desired therapeutic effect in association with the required pharmaceutical vehicle. The dosage unit forms of the disclosure are dictated by and directly dependent on (a) the unique characteristics of the therapeutic composition and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding/formulating such a therapeutic composition for the treatment of a disease or disorder in a patient. In certain embodiments, the compositions of the disclosure are administered to the patient in dosages that range from one to five times per day or more. In other embodiments, the compositions of the disclosure are administered to the patient in range of dosages that include, but are not limited to, once every day, every two days, every three days to once a week, and once every two weeks. It will be readily apparent to one skilled in the art that the frequency of administration of the various combination compositions of the disclosure will vary from subject to subject depending on many factors including, but not limited to, age, disease or disorder to be treated, gender, overall health, and other factors. Thus, the 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 - 73 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) about 1,000 mg, about 10 mg to about 750 mg, about 20 mg to about 600 mg, about 30 mg to about 500 mg, about 40 mg to about 400 mg, about 50 mg to about 300 mg, about 60 mg to about 250 mg, about 70 mg to about 200 mg, about 80 mg to about 150 mg, and any and all whole or partial increments there-in-between. In some embodiments, the dose of active agent (i.e., nucleic acid) present in the composition of the disclosure is from about 0.5 µg and about 5,000 mg. In some embodiments, a dose of active agent present in the composition of the disclosure used in compositions described herein is less than about 5,000 mg, or less than about 4,000 mg, or less than about 3,000 mg, or less than about 2,000 mg, or less than about 1,000 mg, or less than about 800 mg, or less than about 600 mg, or less than about 500 mg, or less than about 200 mg, or less than about 50 mg. Similarly, in some embodiments, a dose of a second compound as described herein is less than about 1,000 mg, or less than about 800 mg, or less than about 600 mg, or less than about 500 mg, or less than about 400 mg, or less than about 300 mg, or less than about 200 mg, or less than about 100 mg, or less than about 50 mg, or less than about 40 mg, or less than about 30 mg, or less than about 25 mg, or less than about 20 mg, or less than about 15 mg, or less than about 10 mg, or less than about 5 mg, or less than about 2 mg, or less than about 1 mg, or less than about 0.5 mg, and any and all whole or partial increments thereof. In certain embodiments, the present disclosure is directed to a packaged pharmaceutical composition comprising a container holding a therapeutically effective amount of the composition of the disclosure, alone or in combination with a second pharmaceutical agent; and instructions for using the compound to treat, prevent, or reduce one or more symptoms of a disease or disorder in a patient. The term "container" includes any receptacle for holding the pharmaceutical composition or for managing stability or water uptake. For example, in certain embodiments, the container is the packaging that contains the pharmaceutical composition, such as liquid (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 - 74 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) functional relationship to the packaged product. However, it should be understood that the instructions may contain information pertaining to the compound's ability to perform its intended function, e.g., treating, preventing, or reducing a disease or disorder in a patient. Administration Routes of administration of any of the compositions of the disclosure include inhalational, oral, nasal, rectal, parenteral, sublingual, transdermal, transmucosal (e.g., sublingual, lingual, (trans)buccal, (trans)urethral, vaginal (e.g., trans- and perivaginally), (intra)nasal, and (trans)rectal), intravesical, intrapulmonary, intraduodenal, intragastrical, intrathecal, epidural, intrapleural, intraperitoneal, subcutaneous, intramuscular, intradermal, intra-arterial, intravenous, intrabronchial, inhalation, and topical administration. Suitable compositions and dosage forms include, for example, tablets, capsules, caplets, pills, gel caps, troches, emulsions, dispersions, suspensions, solutions, syrups, granules, beads, transdermal patches, gels, powders, pellets, magmas, lozenges, creams, pastes, plasters, lotions, discs, suppositories, liquid sprays for nasal or oral administration, dry powder or aerosolized formulations for inhalation, compositions and formulations for intravesical administration and the like. It should be understood that the formulations and compositions that would be useful in the present disclosure are not limited to the particular formulations and compositions that are described herein. Parenteral Administration As used herein, "parenteral administration" of a pharmaceutical composition includes any route of administration characterized by physical breaching of a tissue of a subject and administration of the pharmaceutical composition through the breach in the tissue. Parenteral administration thus includes, but is not limited to, administration of a pharmaceutical composition by injection of the composition, by application of the composition through a surgical incision, by application of the composition through a tissue-penetrating non-surgical wound, and the like. In particular, parenteral administration is contemplated to include, but is not limited to, intracerebroventricular, subcutaneous, intravenous, intraperitoneal, intramuscular, intrasternal injection, and kidney dialytic infusion techniques. In certain embodiments, the composition of the present disclosure is administered intracerebroventricularly. 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 - 75 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) in a form suitable for bolus administration or for continuous administration. Injectable formulations may be prepared, packaged, or sold in unit dosage form, such as in ampules or in multidose containers containing a preservative. Injectable formulations may also be prepared, packaged, or sold in devices such as patient-controlled analgesia (PCA) devices. Formulations for parenteral administration include, but are not limited to, suspensions, solutions, emulsions in oily or aqueous vehicles, pastes, and implantable sustained-release or biodegradable formulations. Such formulations may further comprise one or more additional ingredients including, but not limited to, suspending, stabilizing, or dispersing agents. In certain embodiments of a formulation for parenteral administration, the active ingredient is provided in dry (i.e., powder or granular) form for reconstitution with a suitable vehicle (e.g., sterile pyrogen-free water) prior to parenteral administration of the reconstituted composition. The pharmaceutical compositions may be prepared, packaged, or sold in the form of a sterile injectable aqueous or oily suspension or solution. This suspension or solution may be formulated according to the known art, and may comprise, in addition to the active ingredient, additional ingredients such as the dispersing agents, wetting agents, or suspending agents described herein. Such sterile injectable formulations may be prepared using a non- toxic parenterally acceptable diluent or solvent, such as water or 1,3-butanediol, for example. Other acceptable diluents and solvents include, but are not limited to, Ringer's solution, isotonic sodium chloride solution, and fixed oils such as synthetic mono- or di-glycerides. Other parentally-administrable formulations which are useful include those which comprise the active ingredient in microcrystalline form in a recombinant human albumin, a fluidized gelatin, in a liposomal preparation, or as a component of a biodegradable polymer system. Compositions for sustained release or implantation may comprise pharmaceutically acceptable polymeric or hydrophobic materials such as an emulsion, an ion exchange resin, a sparingly soluble polymer, or a sparingly soluble salt. EXAMPLES Various embodiments of the present application can be better understood by reference 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 TLR7/8 agonist 1 dihydrochloride was purchased from Cayman Chemical (Ann - 76 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) Arbor, MI).1,2-epoxydodecane (C12) and cholesterol was obtained from Sigma Aldrich (St Louis, MO). Core 200 was customized from Enamine (Monmouth Junction, NJ) and other polyamine cores were purchased from Sigma Aldrich and TCI (Tokyo, Japan). Anti-mouse CD16/32 antibody, APC anti-mouse CD11c antibody, FITC anti-mouse CD80 antibody and PE anti-mouse CD86 antibody were purchased from Biolegend (San Diego, CA). Mouse IL-1 beta uncoated ELISA, mouse IL-12 p70 uncoated ELISA, mouse TNF alpha uncoated ELISA, LysoTracker® Deep Red, 3,3’-dioctadecyloxacarbocyanine perchlorate (DiO) and 1,1'-Dioctadecyl-3,3,3',3'-Tetramethylindotricarbocyanine Iodide (DiR) were bought from Invitrogen (Carlsbad, CA).1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2- dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (DMG-PEG 2000) and cholesterol were obtained from Avanti Polar Lipids (Alabaster, AL). DLin-MC3-DMA was purchased from MedChem Express (Monmouth Junction, NJ). m1ψ-modified Luciferase mRNA and SARS-CoV-2 mRNA were produced as previously described in the literature according to methods known to those of ordinary skill in the art. Synthesis of adjuvant lipidoid Adjuvant lipidoid C12-TLRa was synthesized by reacting epoxydodecane (C12) with TLR7/8 agonist 1 dihydrochloride using the ring-opening reaction (i.e., SN2 reaction of an epoxide and amine). Briefly, 10 mg of TLR7/8 agonist 1 dihydrochloride was dissolved in 0.8 mL of ethanol in a glass vial with a magnetic stir bar.8 μL of triethylamine was added to neutralize the hydrochloride before adding 20 mg of C12. The vial was sealed and the mixture was stirred for 48 h at 80 ℃. The crude product was purified by a CombiFlash NextGen 300+ chromatography system (Teledyne ISCO, Lincoln, NE) with gradient elution from CH2Cl2 to 75:22:3 CH2Cl2/MeOH/NH4OH (aq). The desired fraction was collected (yield 44%). C12-TLRa was characterized by mass spectrometry (calculated MS: 728.12, found [M+2H]²⁺ = 365.25) and nuclear magnetic resonance spectroscopy.¹H-NMR (400 MHz, DMSO-d₆) δ: 7.78 (d, J = 8.3 Hz, 1H), 7.57 (dd, J = 8.4, 1.3 Hz, 1H), 7.35 – 7.29 (m, 1H), 7.27 (d, J = 7.9 Hz, 2H), 7.05 – 7.00 (m, 1H), 6.98 (d, J = 8.0 Hz, 2H), 5.84 (s, 2H), 3.61 – 3.47 (m, 2H), 3.44 (s, 2H), 2.90 (t, J = 7.7 Hz, 2H), 2.68 (q, J = 1.9 Hz, 2H), 2.34 (t, J = 2.8 Hz, 2H), 1.69 (p, J = 7.6 Hz, 2H), 1.37 (dt, J = 14.9, 7.5 Hz, 2H), 1.22 (s, 36H), 0.90 – 0.81 (m, 9H). General method for the synthesis of polyamine-derived lipidoids The polyamine cores were reacted with excess moles of C12 needed to saturate the - 77 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) amines as previously described. Take C12-113 as an example, 113 core (1 equiv.) was mixed with C12 (4.8 equiv.) for 48 h at 80 ℃ in a neat condition. The crude product was used for the initial library screening. To purify the top-performing C12-113 lipidoid, the crude product was separated using a CombiFlash NextGen 300+ chromatography system as described above, and the fully saturated product was collected and identified by mass spectrometry _{(calculated MS: 854.49, found [M+2H]} ²⁺ _{= 429.13) and used for subsequent experiments.} Structural simulation of agonist-TLR7 interaction The structures of TLR7/8 agonist 1 and C12-TLRa were first optimized by molecular dynamic simulation at CHARMm force field. The exact TLR7 protein crystal structure was derived from the structural of TLR7/R848 complex (PDB ID: 5GMH) removing any ligands or solvent molecules. Structural simulation between TLR7 dimer and agonists was carried out by CDocker docking simulation and in situ structural superimposition. Potential non-covalent interactions, binding pockets and overviews of the binding sites between TLR7 dimer and the corresponding agonists were generated using BIOVIA Discovery Studio 2018. LNP formulation LNPs were formulated by microfluidic mixing as described previously. Briefly, an ethanol phase containing lipidoid (with or without C12-TLRa substitution), DOPE, cholesterol and DMG-PEG at a molar ratio of 35:16:46.5:2.5 was mixed with an aqueous phase (10 mM citrate buffer, pH 3) containing mRNA at a flow rate of 1:3 and a lipidoid/RNA weight ratio of 10:1 in a microfluidic chip device. LNPs were dialyzed against 1x PBS in a 20 kDa MWCO cassette for 2 h, sterilized through a 0.22 μM filter and stored at 4 ℃. DiO- or DIR-labeled LNPs were obtained by mixing DiO or DiR (1 mol% of total lipids) with LNPs before dialysis. LNP characterization The hydrodynamic size, polydispersity index (PDI) and zeta potential of LNPs were measured using a Zetasizer Nano ZS90 (Malvern Instruments, Malvern, UK). The morphology of LNPs was characterized by transmission electron microscopy (JEOL 1010, Tokyo, Japan). 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. - 78 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) Cell culture and animal studies HEK-Blue mTLR7 cell line was obtained from InvivoGen (#hkb-mtlr7). These cells were maintained according to vendor’s instruction. Murine macrophage DC2.4 cell line was obtained 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. All cells were cultured at 37 ℃ in a humidified incubator of 5% CO2, and routinely tested for mycoplasma contamination. BMDCs were generated from C57BL/6 mice. Briefly, bone marrow cells were flushed from mouse femurs and tibias, lysed by ACK buffer to remove red blood cells, and then cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin and 100 μg/mL streptomycin, 1% HEPES, 0.1% mM β-mercaptoethanol, 20 ng/mL interleukin-4 (IL-4, #214-14, PeproTech) and 20 ng/mL granulocyte-macrophage colony stimulating factor (GM-CSF, #315-03, PeproTech). On day 6, non-adherent and loosely adherent cells were collected for studies. C57BL/6 female mice (6-8 weeks age, 18-20 g body weight) were purchased from Jackson Laboratory. In vitro mLuc delivery DC cells or BMDCs were seed into a 96-well plate at a density of 10,000 per well overnight and then mLuc-loaded LNPs were used to treat cells at the indicated doses for 24 h. Luciferase expression was evaluated by Luciferase Reporter 1000 Assay System (E4550, Promega) and cell viability was measured using a CellTiter-Glo Luminescent Cell Viability Assay (G7572, Promega) according to manufacturer’s protocols. The relative luciferase expression was reported as relative light unit (RLU) normalized to cell viability. Free mRNA was used as a control. TLR7 reporter assay The TLR7-agonistic activity of C12-TLRa was tested on HEK-Blue mTLR7 reporter cells using a HEK-Blue detection kit (#hb-det2, InvivoGen) according to the manufacturer’s instructions. TLR7/8 agonist 1 was used as a positive control. Similarly, the TLR7-agonistic activity of C12-113 LNP or C12-113/TLRa LNP was tested. Cellular uptake DC2.4 cells were seeded into 35-mm glass-bottom dishes for 24 h and then treated - 79 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) with DiO-labelled C12-113 LNP or DiO-labelled C12-113/TLRa LNP at an mRNA concentration of 500 ng/mL for 2 h. Cells were sequentially stained with LysoTracker Deep Red (100 nM) for 30 min and Hoechst 33342 (10 μg/mL) for 5 min. Images were taken immediately using a confocal laser scanning microscope (LSM 710, Zeiss). Analysis of DC maturation and cytokine production in vitro D_{C2.4 cells were seeded in a 12-well plate at a density of 1 x 10} ⁶ _{cells/well overnight} and then treated with SARS-CoV-2 mRNA-loaded LNPs (500 ng/mL) for 24 h. Cell cultures were collected for ELISA analysis of TNF-α, IL-12p70 and IL-1β. Cells were collected, blocked with anti-mouse CD16/32 antibody, and then stained with FITC anti-mouse-CD80 antibody and PE anti-mouse-CD86 antibody for 30 min at 4 ℃ before analyzed by flow cytometry (BD, LSR II). BMDCs were seeded in a 12-well plate at a density of 1 x 10⁶ cells/well and incubated with SARS-CoV-2 mRNA-loaded LNPs (500 ng/mL) for 24 h. Cell cultures were collected for ELISA analysis. Cells were collected, blocked with anti-mouse CD16/32 antibody, and then stained with APC anti-mouse CD11c Antibody, FITC anti- mouse-CD8- antibody, and PE anti-mouse-CD86 antibody for 30 min at 4 ℃ before analysis by flow cytometry. Analysis of DC maturation and cytokine production in vivo Two iLNs from each mouse were harvested at 24 h post-injection of SARS-CoV-2 mRNA-loaded LNPs (5 μg mRNA/mouse) at the tail base and were gently mechanically disrupted using sterile pestles in 0.1 mL of RPMI complete medium in a 1.5 mL tube. The resulting cell suspensions were collected, blocked with anti-mouse CD16/32 antibody, and then stained with APC anti-mouse CD11c Antibody, FITC anti-mouse-CD80 antibody and PE anti-mouse-CD86 antibody for 30 min at 4 ℃ before analyzed by flow cytometry. Blood was collected into serum separator tubes (BD #365967) through the retro- orbital route at 6 and 24 h post immunization. Serum was separated from blood following an incubation period of 30 minutes at room temperature, and samples were centrifuged at 10,000 g for 5 minutes. The serum was stored at -20 ℃ until use. To analyze the intralymphatic cytokine production, the resulting cell suspensions from iLNs were placed to a 96-well plate at a density of 10,000 cells/100 μL/well and cultured for 8 h. Supernatant was collected for ELISA analysis of TNF-α, IL-12p70 and IL-1β together with serum samples. Distribution and transfection of LNPs in vivo - 80 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) Mice were s.c. injected at the tail base with mLuc-loaded LNPs at a dose of 5 μg mRNA per mouse. At 6 h or 24 h post-injection, mice were intraperitoneally (i.p.) injected with D-luciferin potassium salt (150 mg/kg) and bioluminescence imaging was performed on an IVIS imaging system (PerkinElmer). To enable concurrent bioluminescence and fluorescence imaging, DiR-labelled, mLuc-loaded LNPs were injected into mice. At 24 h post-injection, mice were (i.p.) injected with D-luciferin potassium salt, and major organs as well as iLNs were collected for bioluminescence and fluorescence imaging. In vivo immunization Mice were s.c. immunized with SARS-CoV-2 mRNA-loaded LNPs at a dose of 5 μg mRNA per mouse twice using a prime-boost strategy at a three-week interval. Body weight was recorded twice a week during the experiment. Serum was collected using serum separator tubes as described above, stored at -20 ℃ and used for ELISA and virus neutralization assays. Two weeks post the second vaccination, mice were isoflurane- anesthetized and spleens were collected for flow cytometry analysis. Determination of anti-RBD antibody titers using ELISA High Bind Stripwell™ Corning 96 Well Clear Polystyrene Microplates were coated overnight with 1 μg/mL purified SARS-CoV-2 His tagged RBD. Plates were washed once with wash buffer (0.05% Tween-20 in PBS), and blocked for two hours at room temperature using a solution of heat inactivated, IgG depleted, protease free bovine serum albumin (2% w/v BSA in PBS). After blocking, plates were washed three times, and mouse sera was serially diluted in the blocking solution and incubated for 2 h at room temperature. Plates were washed three times before the addition of HRP conjugated anti-mouse secondary antibody specific to total IgG (1:10,000) or subclasses (IgG1: 1:10,000, IgG2c: 10,000) in blocking buffer. Plates were incubated for 1.5 h, washed three times before the addition of 100 µL per well of KPL TMB substrate for 8 min. The reaction was stopped by adding 50 µL of 2N sulfuric acid, and the absorbance was measured at 450 nm using a SpectraMax™ 190 microplate reader. RBD-specific IgG end-point dilution titer was defined as the highest dilution of serum to give an OD greater than the cut-of OD value determined using the Frey method. Pseudovirus neutralization assay Production of VSV pseudotype with SARS-CoV-2 S was performed as described. - 81 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) Antibody neutralization assay using VSVΔG-RFP SARS-CoV-2: Vero E6 cells stably expressing TMPRSS2 were seeded in 100 μL at 2.5 x 10⁴ cells/well in a 96-well collagen coated plate. The next day, 2-fold serially diluted serum samples were mixed with VSVΔG- RFP SARS-CoV-2 pseudotype virus (50-200 focus forming units/well) encoding the spike of D614G, Beta or Delta variant and incubated for 1 hour at 37 °C. Also included in this mixture to neutralize any potential VSV-G carryover virus was 8G5F11, a mouse anti-VSV Indiana G, at a concentration of 100 ng/ml (Absolute Antibody #Ab01401-2.0). The antibody-virus mixture was then used to replace the media on VeroE6 TMPRSS2 cells.20 hours post infection, the cells were washed and fixed with 4% PFA before visualization on an S6 FluoroSpot Analyzer (CTL, Shaker Heights OH). Individual infected foci were enumerated and the values compared to control wells without antibody. The focus reduction neutralization titer 50% (FRNT₅₀) was measured as the greatest serum dilution at which focus count was reduced by at least 50% relative to control cells that were infected with pseudotype virus in the absence of mouse serum. FRNT₅₀ titers for each sample were measured in two technical replicates performed on separate days. Flow cytometry analysis of T cells and B cells T cell. Spleens were collected, processed, as single cells, filtered using a 70 µm cell strainers in complete RPMI-1640, centrifuged, and red blood cells lysed in ACK lysis buffer to obtain a clear single cell suspension. To measure antigen-specific T cells, 2 million splenocytes were stimulated with 2.5 µg/mL of SARS-CoV-2 spike peptide pools (JPT PM- WCPV-S) in a FACS tube for 6 h at 37 ℃, 5% CO2 with 2 mg/ml anti-CD28 (Tonbo #40- 0281-M001) providing co-stimulation. Stimulations proceeded for 1 h before adding 5 mg/mL brefeldin A (Biolegend #420601), 2 mM monensin (Biolegend #420701), and 5 mg/ml anti-CD107a Alexa Fluor 647 (Biolegend #121610) for 5 h. DMSO served as a negative control and the combination of 50 mg/ml phorbol 12-myristate 13-acetate and 1 mg/ml ionomycin served as a positive control. After a total of 6 h, samples were washed with PBS, stained with Live/Dead Aqua for 5 minutes, blocked using anti-mouse CD16/32 antibody for 20 min, and strained extracellularly for 30 min using antibodies (FIG.21D). Cells were washed in FACS buffer, fixed and permeabilized using the Cytofix/Cytoperm kit (BD Biosciences #554714), and stained intracellularly using antibodies for 30 min (FIG. 21D). After intracellular staining, cells were washed twice, fixed with 300 µL (1% paraformaldehyde) and samples were acquired on a BD LSR II equipped with 4 laser lines and 18 PMTs. The gating strategy (FIGs.21A-21C), as well as the antibody list and - 82 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) fluorophore information is provided in Table 1. Table 1. Antibody panel for T cells Marker Fluorophore CD107a FITC CD4 PerCP-Cy5.5 CD8a Pac Blue Aqua live/dead - CD3e BV605 IL-4 BV786 IL-2 APC IFN-g AF700 IL-17 APC/Cy7 IL-5 PE TNF-a PE/Cy7 Memory B cells. Spleens were collected, processed as single cells, filtered using a 40 µm cell strainers in complete RPMI-1640, centrifuged at 300g for 5 minutes, and red blood cells lysed with ACK (1 min), washed twice, counted and 2 million cells per sample incubated with anti-mouse CD16/32 antibody for 20 min at 4 °C. Cells were then washed with FACS buffer (1% BSA in PBS), and stained for 1 h using the panel in (Table 2). Following staining, cells were washed twice, fixed with 300 µL (1% paraformaldehyde) and samples were acquired on a BD LSR II equipped with 4 laser lines and 18 PMTs. The gating strategy, as well as the antibody list, in-house fluorescent RBD probes are provided herein (FIGs.23A-23C and Table 2). Table 2. Antibody panel for T cells Marker Fluorophore Aqua live/dead - CD80 BV650 IgD BV711 B220 BV785 CD38 AF700 - 83 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) PD-L2 PE IgM PECF594 CD4 PECy5 CD8 PECy5 Gr-1 PECy5 F4/80 PECy5 CD19 PECy5.5 GL7 PECy7 RBD BV421 RBD AF647 ELISpot assay Bone marrow was flushed from femurs and tibia into FACS buffer and filtered through 63 µm Nitex mesh. Red blood cells were lysed in ACK buffer for 5 minutes on ice, and washed twice with FACS buffer. Resulting cells were counted using a Beckman Coulter ViCell. MultiScreenHTS IP Filter Plate, 0.45 µm (Millipore Sigma, Catalog: MSIPS4W10) were coated with RBD protein antigen at 10 μg/mL in Sodium Carbonate/Sodium Bicarbonate buffer pH 9.6 (35 mM NaHCO₃ and 15 mM Na₂CO₃) for one hour at 37 ℃. Plates were then washed with 200 µL PBS/ well 3 times and blocked at 37 ℃ in complete RPMI for 30 min. Bone marrow cells were plated in 6 halving dilutions beginning with 1 million total BM cells per well and incubated overnight in complete RPMI. Plates were then washed with wash buffer (1x PBS+0.1% Tween 20) 5 times and biotinylated anti-IgG detection antibody (Goat Anti-Mouse IgG Human ads-BIOT: Southern Biotech cat# 1030-08) was added at final dilution of 3 μg/mL in PBS+2%BSA and incubated at room temperature for one hour. Plates were once again washed 5 times and streptavidin-alkaline phosphatase (1:20000 dilution in PBS+2%BSA) was added prior to incubation at RT for 30 minutes. Plates were then washed 5 times with wash buffer and 50 µL/well BCIP/NBT single solution (Sigma B1911-100 mL) was added for ~10 minutes or until spots developed at which time the reaction was quenched with 100 µL 1M sodium phosphate monobasic solution. After plates were rinsed with deionized H2O and dried overnight they were scanned and counted using CTL Immunospot hardware and software. Statistical analysis - 84 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) 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. Example 1: Synthesis of adjuvant lipidoid and polyamine-derived lipidoids Adjuvant lipidoid C12-TLRa was synthesized by the ring-opening reaction between amine-containing TLR7/8 agonist 1 and C12 epoxide (FIG.2A). The purity and structure of C12-TLRa were verified by liquid chromatography-mass spectrometry and proton nuclear magnetic resonance. Structural simulation via computational analysis demonstrated that C12- _{TLRa could form multiple interactions with TLR7 (e.g., hydrogen bonding with Asp} ⁵⁵⁵ _and Thr⁵⁸⁶ and π-π stacking with Phe⁴⁰⁸) at the first binding site similar to TLR7/8 agonist 1 (FIGs.7A-7F), suggesting that such modifications cause minimal interruption, if any, regarding the agonist-receptor interaction. The TLR7-agonistic activity of C12-TLRa was further verified on HEK-Blue reporter cells stably expressing the TLR7 receptor. As a positive control, free TLR7/8 agonist 1 exhibited a bimodal dose-response profile of TLR7- agonistic activity characterized by an initial dose-dependent increase followed by a decrease of analytes (FIG.8). However, monotonically dose-dependent TLR7-agonistic activity of C12-TLRa was observed at the same dose range, indicating that it has a more predictable pharmaceutical property. Next, adjuvant lipidoid, DOPE, cholesterol and DMG-PEG, along with mRNA, were formulated into C12-TLRa LNP at a molar ratio of 35:16:46.5:2.5 (lipidoid:DOPE:chol:DMG-PEG) using microfluidic mixing. The mRNA encapsulation efficiency was approximately 75%, suggesting that this adjuvant lipidoid is able to complex and encapsulate negatively charged mRNA into LNP like other ionizable lipidoids. m1ψ- modified firefly luciferase mRNA (mLuc)-encapsulated C12-TLRa LNP was then used to treat DC2.4 dendritic cells. While free mLuc was unable to transfect cells, C12-TLRa LNP demonstrated dose-dependent mRNA transfection (FIG.2B), demonstrating that adjuvant lipidoid can successfully deliver mRNA into cells. Table 3. Composition of exemplary LNPs LNP Ionizable lipid Helper lipid Chol.^a PCL^b C12-TLRa C12-TLRa DOPE Cholesterol DMG-PEG - 85 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) (35 mol%) (16 mol%) (46.5 mol%) (2.5 mol%) C12-113 DOPE Cholesterol DMG-PEG C12-113 (35 mol%) (16 mol%) (46.5 mol%) (2.5 mol%) C12- C12-113:C12-TLRa DOPE Cholesterol DMG-PEG 113/TLRa (30:5 mol%) (16 mol%) (46.5 mol%) (2.5 mol%) MC3/DSPC DSPC Cholesterol DMG-PEG MC3 (50 mol%) (10 mol%) (38.5 mol%) (1.5 mol%) MC3/C12-TLRa DSPC Cholesterol DMG-PEG MC3/TLRa (45:5 mol%) (10 mol%) (38.5 mol%) (1.5 mol%) To further evaluate the potency of C12-TLRa, the LNP was compared with a series of polyamine-derived lipidoids synthesized utilizing analogous methods (FIG.2C ). In vitro mLuc transfection results demonstrated that C12-TLRa had a relatively low transfection ability especially when compared to the top performer C12-113 (FIG.2D). Example 2: Optimization and characterization of adjuvant lipidoid substituted LNPs Since abundant antigen expression is favorable for mRNA vaccines, the partial lipidoid in C12-113 LNP (i.e., C12-113) was replaced with C12-TLRa so as to provide the LNP with TLR7/8-agonistic properties without compromising its potency in mRNA delivery. Therefore, a series of LNPs having increasing ratios of C12-113 to TLRa LNP C12-113 (e.g., 1 to 17.5 mol%) were formulated and subjected to in vitro mLuc delivery. Interestingly, along with the increased substitution with C12-TLRa, the mRNA transfection efficiency of C12-113/TLRa LNP first increased and then decreased (FIG.2E). The highest transfection was achieved by C12-113/TLRa LNP with 5 mol % of C12-TLRa, which was chosen for subsequent studies. The adjuvant activity of mRNA-loaded C12-113/TLRa LNP was then verified in HEK-Blue reporter cells. While C12-113 LNP possessed no TLR7-agonistic activity, C12- 113/TLRa LNP exhibited dose-dependent TLR7-agonistic activity (FIG.2F). Consequently, C12-113/TLRa LNP stimulated pro-inflammatory cytokine TNF-α production in DC2.4 cells in a dose-dependent manner, which significantly outperformed C12-113 LNP (FIG.9). To further confirm the general applicability of this strategy, DLin-MC3-DMA (MC3) LNP and adjuvant lipidoid-substituted MC3 LNP (i.e., MC3/TLRa LNP) were formulated and tested. Consistently, MC3/TLRa LNP, but not MC3 LNP, showed TLR7-agonistic activity (FIG. - 86 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) 10). These results demonstrate that the substitution of partial lipidoid with C12-TLRa provides LNPs with adjuvant activity. Next, the physicochemical properties of C12-113/TLRa LNP were characterized. The LNP was found to have a hydrodynamic diameter of approximately 52 nm and a polydispersity index (PDI) of 0.127. Additionally, this LNP had a neutral surface charge and a pKa of 6.42, which meet the criteria for potent in vivo mRNA delivery. The encapsulation efficiency of mRNA in C12-113/TLRa LNP was above 93%. The transmission electron microscopy (TEM) image showed that C12-113/TLRa LNP had a uniform spherical structure (FIG.2G). C12-113 LNP without C12-TLRa substitution had very similar parameters (FIG. 11), suggesting that substitution with 5% C12-TLRa did not change the LNP physicochemical properties. Table 4. Characterization data for exemplary LNPs Diameter Zeta potential Encapsulation LNP PDI pKa (nm) (mV) Efficiency (%) C12-TLRa 62.3 ± 2.1 0.134 0.84 ± 3.1 74.5 ± 1.6 7.15 C12-113 47.2 ± 0.6 0.123 -1.4 ± 5.1 93.5 ± 0.3 6.53 C12- 51.8 ± 1.1 0.127 0.51 ± 4.2 93.4 ± 0.4 6.42 113/TLRa MC3 60.2 ± 1.7 0.118 -1.4 ± 3.4 96.5 ± 0.3 6.60 MC3/TLRa 63.8 ± 0.9 0.108 0.66 ± 3.0 96.0 ± 0.2 6.61 Example 3: In vitro mRNA delivery and DC activation The in vitro mRNA delivery and adjuvant activity of C12-113/TLRa LNP was next systematically investigated. Both C12-113 LNP and C12-113/TLRa LNP exhibited dose- dependent mLuc transfection in DC2.4 cells (FIG.3A), however, in line with FIG.2E, C12- 113/TLRa LNP consistently outperformed C12-113 LNP at any mRNA dose tested. No major cytotoxicity was observed for both LNPs (FIGs.12A-12B). Similarly, in primary bone marrow-derived dendritic cells (BMDCs), C12-113/TLRa LNP consistently exhibited higher mRNA transfection efficiency than C12-113 LNP (FIG.3B). Accordantly, MC3/TLRa significantly outperformed MC3 LNP in mLuc delivery as well (FIG.13). These results strongly confirm that C12-TLRa substitution can increase LNP-mediated mRNA delivery in vitro. - 87 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) In order to investigate the improved mRNA transfection following C12-TLRa incorporation into LNP, the endosomal escape of LNPs was evaluated. Lipid-like fluorescent dye DiO-labelled LNPs were used to treat DC2.4 cells and their subcellular distribution was visualized after endo/lysosomes and nuclei were counterstained (FIG.3C). In C12-113 LNP- treated cells, there were massive yellow spots, indicating severe entrapment of C12-113 LNP (green) inside endo/lysosomes (red). In contrast, C12-113/TLRa LNP-treated cells exhibited more homogenous green signal across the cytoplasm, suggesting a more efficient escape from endo/lysosomes. These results suggest that C12-TLRa substitution enhances endosomal escape of LNP, which could explain the improved mRNA transfection (FIG.3A-3B). Since multiple interactions are observed between C12-TLRa and TLR7 based on the molecular simulation result (FIGs.7A-7E), it is reasonable to assume that the strong affinity between the incorporated C12-TLRa and endosomal TLR7/8 could reinforce the physical interaction between LNP and endosomal membrane, which leads to the enhanced endosomal disruption (FIG.3D). Next, the immunostimulatory effect of C12-113/TLRa LNP carrying m1ψ-modified SARS-CoV-2 mRNA encoding the diproline modified spike glycoprotein of SARS-CoV-2 was evaluated. Notably, the coding sequence of this nucleoside-modified mRNA is identical to the mRNA used in two FDA-approved vaccines (i.e., mRNA-1273 and BNT162b2). Since TLR7/8 agonist can stimulate DC maturation and pro-inflammatory cytokine release, _{matured DCs (CD80} ⁺ _CD86 ⁺ _{) were analyzed using flow cytometry and secreted TNF-α, IL-} 12p70, and IL-1β using enzyme-linked immunosorbent assay (ELISA) at 24 h post-treatment. C12-113/TLRa LNP markedly increased the percentage of matured DCs in both DC2.4 cells and BMDCs (FIGs.3E-3F and FIGs.14A-14B). In contrast, C12-113 LNP modestly induced DC maturation. Moreover, C12-113/TLRa LNP stimulated a significant increase in TNF-α, IL-12p70 and IL-1β levels compared to C12-113 LNP in both DC2.4 cells and BMDCs (FIGs.3G-3H). Taken together, these results suggest that C12-TLRa substitution greatly enhances the adjuvanticity of LNPs and DC activation. Example 4: In vivo mRNA transfection and innate immune responses Next, it was examined whether or not increased mRNA transfection and innate immune responses by adjuvant lipidoid-substituted LNPs could reproduced in vivo. C57BL/6 mice were subcutaneously (s.c.) immunized at the tail bases of mice with mLuc-loaded C12- 113 LNP or C12-113/TLRa LNP, and luciferase expression was visualized by in vivo bioluminescence imaging at 6 and 24 h post-treatment (FIG.4A). Both LNPs mainly - 88 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) transfected injection sites, and could also transfect inguinal lymph nodes (iLNs). In line with in vitro transfection results, C12-113/TLRa LNP achieved greater mRNA transfection than C12-113 LNP at both injection sites and iLNs (FIG.4A). To avoid systemic toxicity, it is important to spatially restrict adjuvant activity and mRNA transfection to the sites of vaccine administration and draining lymph nodes. Therefore, the distribution and transfection of LNPs in major organs and iLNs was studied at 24 h post-injection of lipid-like fluorescent dye DiR-labelled, mLuc-loaded LNPs. Ex vivo fluorescence and luminescence imaging confirmed that no obvious accumulation and transfection in major organs (i.e., liver, heart, spleen, lung, and kidney) were observed for both LNPs (FIG.4B). Interestingly, although the two LNPs achieved comparable accumulation in iLNs based on the fluorescence quantification (FIG.4C), C12-113/TLRa LNP enabled stronger mRNA transfection in iLNs than C12-113 LNP, which was in line with in vivo ventral luminescence imaging (FIG.4A). These results also correlate well with in vitro mRNA transfection (FIGs.3A-3B). Moreover, C12-113/TLRa LNP-mediated enhanced mRNA expression at both the injection site and iLNs were highly reproducible from different LNP batches (FIG.15). Finally, the duration and translational kinetics of LNP-formulated mRNA was investigated. C12-113/TLRa LNP enabled more durable (over 14 days) and consistently stronger mRNA expression at the injection site than C12-113 LNP (FIGs.16A-16B). Such long-lasting expression is attributed to both nucleoside modification and LNP formulation. Together, these results suggest that C12-TLRa substitution can greatly enhance the expression of LNP-formulated mRNA at both the injection site and draining LNs with minimal systemic off-target distribution or expression. To assess the in vivo innate immune responses, iLNs were harvested from mice at 24 h post-immunization with SARS-CoV-2 mRNA-loaded LNPs, and intralymphatic DC maturation was analyzed by flow cytometry. C12-113/TLRa LNP dramatically increased the percentage of matured DCs (FIGs.4D-4E and FIG.17), which was significantly higher than C12-113 LNP. Next, iLNs and serum cytokines (TNF-α, IL-12p70 and IL-1β) were analyzed by ELISA at 6 and 24 h post-vaccination. While C12-113 LNP moderately and transiently induced intralymphatic cytokine production, C12-113/TLRa LNP elicited higher-magnitude and more persistent cytokine responses (FIGs.4F-4H). As expected, due to minimal systemic exposure of LNPs (FIGs.4B-4C), serum cytokine levels did not increase for both LNPs (FIGs.18A-18C). These results demonstrate that C12-TLRa substitution can greatly enhance - 89 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) the magnitude and duration of local innate immune responses triggered by SARS-CoV-2 mRNA-LNP vaccine without inducing systemic inflammation. Example 5: Enhanced cellular immune responses against SARS-CoV-2 The adaptive immune responses elicited by original and adjuvant lipidoid-substituted SARS-CoV-2 mRNA-LNP vaccines was further investigated. Mice were vaccinated twice using a prime-boost strategy at a three-week interval (FIG.5A). Of note, neither skin abnormalities at the injection sites (FIG.19) nor body weight loss was observed after immunization (FIG.20). Two weeks after the boost dose, spleens from immunized mice were collected, and splenocytes stimulated with SARS-CoV-2 receptor binding domain (RBD) peptide pools. Antigen-specific CD4⁺ and CD8⁺ T cells were measured by flow cytometry _{(FIGs. 5B-5C and FIGs. 21A-21C). Both LNPs elicited RBD-specific CD4} ⁺ _{T cells} expressing Th1 cytokines (interferon [IFN]-γ, IL-2 and TNF-α), however, the magnitude of C12-113/TLRa LNP was significantly higher compared to C12-113 LNP (FIG.5B). _{Moreover, neither vaccine formulation induced CD4} ⁺ _{T cells expressing type 2 (Th2)} cytokines (IL-4, IL-5 and IL-17), supporting a Th1-biased T cell immune response. C12- _{113/TLRa LNP also elicited more RBD-specific CD8} ⁺ _{T cells expressing Th1 immune} response cytokines (IFN-γ, IL-2 and TNF-α) and cytotoxic marker (CD107α) (FIG.5C). These results suggest that C12-TLRa-substituted SARS-CoV-2 mRNA-LNP vaccines _{can induce stronger RBD-specific Th1 and CD8} ⁺ _{T cell immune responses. Polyfuncitonal T} cells are recognized as a better immune correlate for protection against pathogens. Next, the polyfunctionality of these RBD-specific T cell responses were evaluated. C12-113/TLRa LNP induced a significantly higher proportion of double and triple positive CD4⁺ and CD8⁺ T cells compared to the C12-113 LNP (FIG.5D). The pattern of cytokines secretion of the _{double positive cells was different between CD4} ⁺ _(IL-2 ⁺ _TNF-α ⁺ _{) and CD8} ⁺ _(INF-γ ⁺ _TNF-α ⁺ ₎ cells and consistent with the role of the helper and cytotoxic activities of these two different lymphocyte populations. Example 6: Enhanced humoral immune responses against SARS-CoV-2 Next, SARS-CoV-2 mRNA-LNP vaccine-induced humoral immune responses were analyzed (FIG.6A). Serum from vaccinated mice was subjected to endpoint dilution ELISA to determine the total RBD-specific IgG binding antibody titers. Both mRNA-LNP vaccines triggered high levels of RBD-specific IgG (FIG.6B), however, the IgG titer for C12- - 90 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) 113/TLRa LNP was one-order higher than C12-113 LNP (1.22 x 10⁷ vs.1.35 x 10⁶). Moreover, the ELISA analysis of subsets of RBD-specific IgG showed that C12-113/TLRa LNP elicited similar IgG1 titers and higher IgG2c titers with a higher IgG2c/IgG1 ratio compared to C12-113 LNP (FIG.22), further highlighting that adjuvant lipidoid-substituted LNP tended to trigger stronger Th1-biased immune responses. Afterwards, neutralizing antibody (NAb) levels were measured by a vesicular stomatitis virus (VSV)-based pseudovirus neutralization assay (FIG.6C). Both mRNA-LNP vaccines induced high levels of broad NAbs against SARS-CoV-2 D614G-mutated ancestor strain, beta (B.1.351) and (B.1.617.2) variants. Specifically, the 50% foci reduction neutralization titer (FRNT50) approached ~1/5608 against SARS-CoV-2 beta variant (B.1.351) for the C12-113/TLRa LNP vaccine, which was significantly higher than the FRNT50 of ~1/2640 for the C12-113 LNP vaccine. To investigate the induction of B cell memory, splenic B cell responses were assessed using fluorescent RBD probes (FIGs.23A-23C). C12-113/TLRa LNP vaccination significantly increased the number of RBD-specific B cells compared to C12-113 LNP (FIG. 6D), and these cells displayed a memory phenotype (CD38⁺GL7-; FIG.6E). The majority of RBD-specific B cells in C12-113/TLRa LNP-vaccinated mice co-expressed the memory markers PD-L2 and CD80 (FIG.6E), consistent with a potential for rapid differentiation into antibody-secreting cells (ASCs) upon re-challenge. Thus, C12-113/TLRa LNP vaccination generates a larger but phenotypically similar RBD-specific memory B cell pool. Finally, bone marrow-resident long-lived plasma cells (LLPCs) were analyzed three months after the second vaccination, which can mediate durable protection from infection by persistently producing antigen-specific antibodies. Bone marrow from immunized mice was collected and various subsets of RBD-specific ASCs were characterized by enzyme-linked immunospot assay (ELISPOT). While C12-113 LNP modestly induced the generation of RBD-specific IgG1-, IgG2a-, and IgG2b-expressing ASCs (FIGs.6F-6H), C12-113/TLRa LNP induced 6.7-, 2.4-, and 6.5-fold more IgG₁-, IgG_2a- and IgG_2b-expressing ASCs, respectively. Altogether, these results suggest that adjuvant lipidoid-substituted mRNA-LNP vaccine elicits stronger humoral immune responses and LLPC responses. Enumerated Embodiments The following exemplary embodiments are provided, the numbering of which is not to be construed as designating levels of importance: - 91 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) Embodiment 1 provides a compound of Formula (I), or a salt, solvate, stereoisomer, or isotopologue thereof: , wherein: R¹ is selected from the substituted C₁-C₆ alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C₆-C₁₀ aryl, and optionally substituted heteroaryl; R² is selected from the group consisting of H, R⁵, optionally substituted C1-C6 alkyl, optionally substituted C₃-C₈ cycloalkyl, optionally substituted C₂-C₈ heterocycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted heteroaryl; R^3a and R^3b are each independently selected from the group consisting of H, R⁵, R^6a, and optionally substituted C1-C6 alkyl; R^4a, R^4b, R^4c, and R^4d are each independently selected from the group consisting of H, R⁵, halogen, CN, NO2, optionally substituted C1-C6 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C₂-C₈ heterocycloalkyl, optionally substituted C₆-C₁₀ aryl, optionally substituted C2-C10 heteroaryl, OR^A, N(R^A)(R^B), C(=O)N(R^A)(R^B), C(=O)R^A, C(=O)OR^A, OC(=O)R^A, OC(=O)OR^A, SR^A, S(=O)R^A, S(=O)₂R^A, N(R^A)S(=O)₂R^B, or at least one of R^3a and R^3b is R^6a; each occurrence of R^6a and R^6b is independently selected from the group consisting of -(optionally substituted C₁-C₆ alkylenyl)-C(=O)OR^7a, -(optionally substituted C₁-C₆ alkylenyl)-C(=O)N(R^7a)(R^7b), -(optionally substituted C1-C6 alkylenyl)-C(=O)R^7a, - (optionally substituted C₁-C₆ alkylenyl)-(R^7a), -C(=O)OR^7a, -C(=O)N(R^7a)(R^7b), -C(=O)R^7a, and R^7a; each occurrence of R^7a and R^7b is independently selected from the group consisting of optionally substituted C6-C28 alkyl, optionally substituted C6-C28 heteroalkyl, optionally - 92 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) substituted C3-C8 cycloalkyl, optionally substituted C6-C8 heterocycloalkyl, optionally substituted C₆-C₂₈ alkenyl, and optionally substituted C₆-C₂₈ alkynyl; L is selected from the group consisting of a bond, optionally substituted C1-C6 alkylenyl, optionally substituted C₂-C₆ alkenylenyl, optionally substituted C₂-C₆ heteroalkylenyl, optionally substituted C3-C8 cycloalkylenyl, optionally substituted C2-C8 heterocycloalkylenyl, optionally substituted C₇-C₁₂ aralkylenyl, optionally substituted C₅-C₁₂ heteroaralkylenyl, optionally substituted C6-C10 arylenyl, and optionally substituted C2-C10 heteroarylenyl; and each occurrence of R^A and R^B is independently selected from the group consisting of H, C(=O)(optionally substituted C₁-C₆ alkyl), C(=O)(optionally substituted C₃-C₈ cycloalkyl), C(=O)(optionally substituted C2-C8 heterocycloalkyl), C(=O)(optionally substituted C₆-C₁₀ aryl), C(=O)(optionally substituted C₂-C₁₀ heteroaryl), optionally substituted C1-C6 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted C₂-C₁₀ heteroaryl. Embodiment 2 provides the compound of Embodiment 1, wherein exactly one of R¹, R², R^3a, R^3b, R^4a, R^4b, R^4c, and R^4d is R⁵. of Embodiment 1 or 2, wherein the compound of Formula (I) is a compound of Formula (Ia), or a salt, solvate, stereoisomer, or isotopologue thereof: , R² is selected from the substituted C₁-C₆ alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C₆-C₁₀ aryl, and optionally substituted heteroaryl; R^3a and R^3b are each independently selected from the group consisting of H, and optionally substituted C₁-C₆ alkyl; R^4a, R^4b, R^4c, and R^4d are each independently selected from the group consisting of H, halogen, CN, C₁-C₆ alkyl, optionally substituted C₃-C₈ cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C6-C10 aryl, optionally substituted C₂-C₁₀ heteroaryl, OR^A, N(R^A)(R^B), C(=O)N(R^A)(R^B), C(=O)R^A, - 93 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) C(=O)OR^A, OC(=O)R^A, OC(=O)OR^A, SR^A, S(=O)R^A, S(=O)2R^A, N(R^A)S(=O)2R^B, N(R^A)C(=O)R^B, and S(=O)₂N(R^A)(R^B); R^6a and R^6b are each independently selected from the group consisting of -(optionally substituted C₁-C₆ alkylenyl)-C(=O)OR^7a, -(optionally substituted C₁-C₆ alkylenyl)- C(=O)N(R^7a)(R^7b), -(optionally substituted C1-C6 alkylenyl)-C(=O)R^7a, -(optionally substituted C₁-C₆ alkylenyl)-(R^7a), -C(=O)OR^7a, -C(=O)N(R^7a)(R^7b), -C(=O)R^7a, and R^7a; each occurrence of R^7a and R^7b is independently selected from the group consisting of optionally substituted C₁-C₂₈ alkyl, optionally substituted C₂-C₂₈ heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C₂-C₂₈ alkenyl, and optionally substituted C₂-C₂₈ alkynyl; L is selected from the group consisting of a bond, optionally substituted C1-C6 alkylenyl, optionally substituted C₂-C₆ alkenylenyl, optionally substituted C₂-C₆ heteroalkylenyl, optionally substituted C3-C8 cycloalkylenyl, optionally substituted C2-C8 heterocycloalkylenyl, optionally substituted C₇-C₁₂ aralkylenyl, optionally substituted C₅-C₁₂ heteroaralkylenyl, optionally substituted C6-C10 arylenyl, and optionally substituted C2-C10 heteroarylenyl; and each occurrence of R^A and R^B is independently selected from the group consisting of H, C(=O)(optionally substituted C1-C6 alkyl), C(=O)(optionally substituted C3-C8 cycloalkyl), C(=O)(optionally substituted C₂-C₈ heterocycloalkyl), C(=O)(optionally substituted C6-C10 aryl), C(=O)(optionally substituted C2-C10 heteroaryl), optionally substituted C₁-C₆ alkyl, optionally substituted C₃-C₈ cycloalkyl, optionally substituted C₆-C₁₀ aryl, and optionally substituted C2-C10 heteroaryl. Embodiment 4 provides the compound of any one of Embodiments 1-3, wherein R² is n-butyl. Embodiment 5 provides the compound of any one of Embodiments 1-4, wherein R^3a and R^3b are each independently H. Embodiment 6 provides the compound of any one of Embodiments 1-5, wherein at least one of the following applies: (a) at least one of R^4a, R^4b, R^4c, and R^4d is H; (b) at least two of R^4a, R^4b, R^4c, and R^4d are H; (c) at least three of R^4a, R^4b, R^4c, and R^4d are H; (d) each of R^4a, R^4b, R^4c, and R^4d are H. Embodiment 7 provides the compound of any one of Embodiments 1-6, wherein L is optionally substituted C7-C12 aralkylenyl. - 94 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) Embodiment 8 provides the compound of any one of Embodiments 1-7, wherein L is . 9 provides the compound of any one of Embodiments 1-8, wherein R^6a and R^6b are each independently selected from the group consisting of R^7a and - CH₂CH₂C(=O)OR^7a. Embodiment 10 provides the compound of any one of Embodiments 1-9, wherein R^7a and R^7b are each independently selected from the group consisting of optionally substituted C6-C28 alkyl, optionally substituted C6-C28 alkenyl, and optionally substituted C6-C28 heteroalkyl. Embodiment 11 provides the compound of any one of Embodiments 1-10, wherein R^6a and R^6b are each independently selected from the group consisting of - CH2CH(OH)(optionally substituted C6-C28 alkyl), -CH2CH(OH)(optionally substituted C6- C₂₈ alkenyl), -CH₂CH(OH)(optionally substituted C₆-C₂₈ heteroalkyl), - CH2CH2C(=O)O(optionally substituted C6-C28 alkyl), -CH2CH2C(=O)O(optionally substituted C6-C28 alkenyl), and -CH2CH2C(=O)O(optionally substituted C6-C28 heteroalkyl). Embodiment 12 provides the compound of any one of Embodiments 1-11, wherein R^6a and R^6b are each independently selected from the group consisting of - CH₂CH(OH)(CH₂)₉CH₃ and -CH₂CH₂C(=O)O(CH₂)₁₁CH₃. Embodiment 13 provides the compound of any one of Embodiments 1-12, which is selected from the group consisting of: . (a) at least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent; - 95 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) (b) at least one ionizable lipid; (c) at least one helper lipid; (d) cholesterol; and (e) at least one polymer conjugated lipid. Embodiment 15 provides the LNP of Embodiment 14, wherein the hydrocarbyl substituted TLR agonist is a compound of Formula (II): A(B)n (II), wherein: A is a toll-like receptor (TLR) agonist; each occurrence of B is independently selected from the group consisting of R^6a and ; of R^6a and R^6b is independently selected from the group consisting of -(optionally substituted C1-C6 alkylenyl)-C(=O)OR^7a, -(optionally substituted C1-C6 alkylenyl)-C(=O)N(R^7a)(R^7b), -(optionally substituted C1-C6 alkylenyl)-C(=O)R^7a, - (optionally substituted C₁-C₆ alkylenyl)-(R^7a), -C(=O)OR^7a, -C(=O)N(R^7a)(R^7b), -C(=O)R^7a, and R^7a, each occurrence of R^7a and R^7b is independently selected from the group consisting of optionally substituted C1-C28 alkyl, optionally substituted C2-C28 heteroalkyl, optionally substituted C₃-C₈ cycloalkyl, optionally substituted C₂-C₈ heterocycloalkyl, optionally substituted C2-C28 alkenyl, and optionally substituted C2-C28 alkynyl; L is bond, optionally substituted C₁-C₆ alkylenyl, optionally substituted C₂-C₆ alkenylenyl, optionally substituted C2-C6 heteroalkylenyl, optionally substituted C3-C8 cycloalkylenyl, optionally substituted C₂-C₈ heterocycloalkylenyl, optionally substituted C₇- C12 aralkylenyl, optionally substituted C5-C12 heteroaralkylenyl, optionally substituted C6-C10 arylenyl, and optionally substituted C₂-C₁₀ heteroarylenyl; and n is an integer selected from the group consisting of 1, 2, 3, and 4. Embodiment 16 provides the LNP of Embodiment 14 or 15, wherein the TLR agonist is selected from the group consisting of 1-(4-(aminomethyl)benzyl)-2-butyl-1H-imidazo[4,5- c]quinolin-4-amine (TLR7/8 agonist 1), 1-isobutyl-1H-imidazo[4,5-c]quinolin-4-amine (imiquimod), 1-(4-amino-2-(ethoxymethyl)-1H-imidazo[4,5-c]quinolin-1-yl)-2- methylpropan-2-ol (resiquimod), and 1-(4-amino-2-((ethylamino)methyl)-1H-imidazo[4,5- - 96 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) c]quinolin-1-yl)-2-methylpropan-2-ol (gardiquimod). Embodiment 17 provides the LNP of Embodiment 14, wherein the least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent is the compound of Formula (I) of any one of Embodiments 1-13. Embodiment 18 provides the LNP of any one of Embodiments 14-17, wherein the at least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent is selected from the group consisting of: . the ionizable lipid is at least one selected from the group consisting of: , R^8a, R^8b, R^8c, R^8d, R^8e, and R^8f are each independently selected from the group consisting of -(optionally substituted C1-C6 alkylenyl)-C(=O)OR^9a, -(optionally substituted C₁-C₆ alkylenyl)-C(=O)N(R^9a)(R^9b), -(optionally substituted C₁-C₆ alkylenyl)-C(=O)R^9a, - - 97 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) (optionally substituted C1-C6 alkylenyl)-(R^9a), -C(=O)OR^9a, -C(=O)N(R^9a)(R^9b), -C(=O)R^9a, and R^9a; and each occurrence of R^9a and R^9b is independently selected from the group consisting of optionally substituted C₁-C₂₈ alkyl, optionally substituted C₂-C₂₈ heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C₂-C₂₈ alkenyl, and optionally substituted C₂-C₂₈ alkynyl. Embodiment 20 provides the LNP of Embodiment 19, wherein R^8a, R^8b, R^8c, R^8d, R^8e, and R^8f are each independently selected from the group consisting of R^9a and - CH2CH2C(=O)OR^9a. Embodiment 21 provides the LNP of Embodiment 19 or 20, wherein R^9a and R^9b are each independently selected from the group consisting of optionally substituted C1-C28 alkyl, optionally substituted C₁-C₂₈ alkenyl, and optionally substituted C₁-C₂₈ heteroalkyl. Embodiment 22 provides the LNP of any one of Embodiments 19-21, wherein R^7a and R^7b are each independently selected from the group consisting of - CH2CH(OH)(optionally substituted C1-C28 alkyl), -CH2CH(OH)(optionally substituted C1- C28 alkenyl), -CH2CH(OH)(optionally substituted C2-C28 heteroalkyl), - CH2CH2C(=O)O(optionally substituted C1-C28 alkyl), -CH2CH2C(=O)O(optionally substituted C1-C28 alkenyl), and -CH2CH2C(=O)O(optionally substituted C2-C28 heteroalkyl). Embodiment 23 provides the LNP of any one of Embodiments 19-22, wherein R^7a and R^7b are each independently selected from the group consisting of - CH₂CH(OH)(CH₂)₉CH₃ and -CH₂CH₂C(=O)O(CH₂)₁₁CH₃. Embodiment 24 provides the LNP of any one of Embodiments 14-23, wherein the at least one ionizable lipid comprises: . at least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent comprises about 1 mol% to about 17.5 mol% of the LNP. Embodiment 26 provides the LNP of any one of Embodiments 14-25, wherein the at least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent comprises about 1, 2.5, 5, 10, or about 17.5 mol% of the LNP. - 98 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) Embodiment 27 provides the LNP of any one of Embodiments 14-26, wherein the at least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent comprises about 5 mol% of the LNP. Embodiment 28 provides the LNP of any one of Embodiments 14-27, wherein the at least one ionizable lipid comprises about 10 mol% to about 60 mol% of the LNP. Embodiment 29 provides the LNP of any one of Embodiments 14-28, wherein the at least one ionizable lipid comprises about 34, 32.5, 30, 25, or about 17.5 mol% of the LNP. Embodiment 30 provides the LNP of any one of Embodiments 14-29, wherein the at least one ionizable lipid comprises about 30 mol% of the LNP. Embodiment 31 provides the LNP of any one of Embodiments 14-30, wherein the at least one helper lipid comprises at least one selected from the group consisting of dioleoylphosphatidylethanolamine (DOPE) and distearoylphosphatidylcholine (DSPC). Embodiment 32 provides the LNP of any one of Embodiments 14-31, wherein the at least one helper lipid comprises about 1 mol% to about 50 mol% of the LNP. Embodiment 33 provides the LNP of any one of Embodiments 14-32, wherein the at least one helper lipid comprises about 16 mol% of the LNP. Embodiment 34 provides the LNP of any one of Embodiments 14-33, wherein the cholesterol comprises about 5 mol% to about 60 mol% of the LNP. Embodiment 35 provides the LNP of any one of Embodiments 14-34, wherein the cholesterol comprises about 46.5 mol% of the LNP. Embodiment 36 provides the LNP of any one of Embodiments 14-35, wherein the at least one polymer conjugated lipid comprises 1,2-dimyristoyl-rac-glycero-3- methoxypolyethylene glycol-2000 (DMG-PEG 2000). Embodiment 37 provides the LNP of any one of Embodiments 14-36, wherein the at least one polymer conjugated lipid comprises about 0.1 mol% to about 20 mol% of the LNP. Embodiment 38 provides the LNP of any one of Embodiments 14-37, wherein the at least one polymer conjugated lipid comprises about 2.5 mol% of the LNP. Embodiment 39 provides the LNP of any one of Embodiments 14-38, wherein the LNP has a molar ratio of (a) : (b) : (c) : (d) : (e) of about 5 : 30 : 16 : 46.5 : 2.5. Embodiment 40 provides the LNP of any one of Embodiments 14-39, wherein the LNP further comprises at least one cargo molecule. Embodiment 41 provides the LNP of Embodiment 40, wherein the cargo is at least one selected from the group consisting of a nucleic acid, small molecule, protein, therapeutic agent, antibody, and any combinations thereof. - 99 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) Embodiment 42 provides the LNP of Embodiment 40 or 41, wherein the cargo is a nucleic acid. Embodiment 43 provides the LNP of Embodiment 41 or 42, wherein the nucleic acid is DNA or RNA. Embodiment 44 provides the LNP of any one of Embodiments 41-43, wherein the nucleic acid is selected from the group consisting of mRNA, cDNA, pDNA, microRNA, siRNA, modified RNA, antagomir, antisense molecule, and any combinations thereof. Embodiment 45 provides the LNP of any one of Embodiments 40-44, wherein the cargo is at least partially encapsulated in the LNP. Embodiment 46 provides the LNP of any one of Embodiments 40-45, wherein the cargo is mRNA. Embodiment 47 provides the LNP of Embodiment 46, wherein the LNP has a weight ratio of total lipidoid (i.e., hydrocarbyl substituted toll-like receptor agonist and ionizable lipid) to mRNA ranging from about 5:1 to about 20:1 (i.e., weight ratio of (a) + (b) : mRNA). Embodiment 48 provides the LNP of Embodiment 46 or 47, wherein the LNP has a weight ratio of total lipidoid (i.e., hydrocarbyl substituted toll-like receptor agonist and ionizable lipid) to mRNA of about 10:1 (i.e., weight ratio of (a) + (b) : mRNA). Embodiment 49 provides the LNP of any one of Embodiments 44-48, wherein the mRNA encodes SARS-CoV-2, an immunogenic fragment thereof (e.g., spike protein), or a modified derivative thereof. Embodiment 50 provides a pharmaceutical composition comprising the lipid nanoparticle (LNP) of any one of Embodiments 14-49 and a pharmaceutically acceptable carrier. Embodiment 51 provides a method of generating an innate immune response in a subject, the method comprising administering to the subject the lipid nanoparticle (LNP) of any one of Embodiments 14-49 or the pharmaceutical composition of Embodiment 50. Embodiment 52 provides a method of treating, preventing, and/or ameliorating an infection, disease, or disorder in a subject, the method comprising administering to the subject a lipid nanoparticle (LNP) comprising: (a) at least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent; (b) at least one ionizable lipid; (c) at least one helper lipid; (d) cholesterol; - 100 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) (e) at least one polymer conjugated lipid; and (f) at least one cargo molecule. Embodiment 53 provides the method of Embodiment 52, wherein the cargo is a nucleic acid. Embodiment 54 provides the method of Embodiment 52 or 53, wherein the nucleic acid is DNA or RNA. Embodiment 55 provides the method of any one of Embodiments 52-54, wherein the nucleic acid is selected from the group consisting of mRNA, cDNA, pDNA, microRNA, siRNA, modified RNA, antagomir, antisense molecule, and any combinations thereof. Embodiment 56 provides the method of any one of Embodiments 52-55, wherein the cargo is at least partially encapsulated in the LNP. Embodiment 57 provides the method of any one of Embodiments 52-56, wherein the cargo is mRNA. Embodiment 58 provides the method of Embodiment 57, wherein the mRNA encodes SARS-CoV-2, an immunogenic fragment thereof (e.g., spike protein), or a modified derivative thereof. Embodiment 59 provides the method of Embodiment 58, wherein the infection, disease, or disorder is a SARS-CoV-2 infection. Embodiment 60 provides the method of any one of Embodiments 52-59, wherein an innate immune response is promoted in the subject. Embodiment 61 provides the method of any one of Embodiments 51-60, wherein the subject is a mammal. Embodiment 62 provides the method of Embodiment 61, 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. - 101 - 52436892.1

Claims

Attorney Docket No.046483-7434WO1(03633) CLAIMS What is claimed is: 1. A compound of Formula (I), or a salt, solvate, stereoisomer, or isotopologue thereof: , wherein: 1 R is selected from the group substituted C₁-C₆ alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C₆-C₁₀ aryl, and optionally substituted heteroaryl; R² is selected from the group consisting of H, R⁵, optionally substituted C1-C6 alkyl, optionally substituted C₃-C₈ cycloalkyl, optionally substituted C₂-C₈ heterocycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted heteroaryl; R^3a and R^3b are each independently selected from the group consisting of H, R⁵, R^6a, and optionally substituted C1-C6 alkyl; R^4a, R^4b, R^4c, and R^4d are each independently selected from the group consisting of H, R⁵, halogen, CN, NO2, optionally substituted C1-C6 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C₂-C₈ heterocycloalkyl, optionally substituted C₆-C₁₀ aryl, optionally substituted C2-C10 heteroaryl, OR^A, N(R^A)(R^B), C(=O)N(R^A)(R^B), C(=O)R^A, C(=O)OR^A, OC(=O)R^A, OC(=O)OR^A, SR^A, S(=O)R^A, S(=O)2R^A, N(R^A)S(=O)2R^B, R⁵ is , wherein at least one of R¹, R², R^3a, R^3b, R^4a, R^4b, R^4c, and R^4d is R⁵, or at least one of R^3a and R^3b is R^6a; each occurrence of R^6a and R^6b is independently selected from the group consisting of -(optionally substituted C₁-C₆ alkylenyl)-C(=O)OR^7a, -(optionally substituted C₁-C₆ alkylenyl)-C(=O)N(R^7a)(R^7b), -(optionally substituted C1-C6 alkylenyl)-C(=O)R^7a, - (optionally substituted C₁-C₆ alkylenyl)-(R^7a), -C(=O)OR^7a, -C(=O)N(R^7a)(R^7b), -C(=O)R^7a, and R^7a; - 102 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) each occurrence of R^7a and R^7b is independently selected from the group consisting of optionally substituted C₆-C₂₈ alkyl, optionally substituted C₆-C₂₈ heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C6-C8 heterocycloalkyl, optionally substituted C₆-C₂₈ alkenyl, and optionally substituted C₆-C₂₈ alkynyl; L is selected from the group consisting of a bond, optionally substituted C1-C6 alkylenyl, optionally substituted C₂-C₆ alkenylenyl, optionally substituted C₂-C₆ heteroalkylenyl, optionally substituted C3-C8 cycloalkylenyl, optionally substituted C2-C8 heterocycloalkylenyl, optionally substituted C₇-C₁₂ aralkylenyl, optionally substituted C₅-C₁₂ heteroaralkylenyl, optionally substituted C6-C10 arylenyl, and optionally substituted C2-C10 heteroarylenyl; and each occurrence of R^A and R^B is independently selected from the group consisting of H, C(=O)(optionally substituted C₁-C₆ alkyl), C(=O)(optionally substituted C₃-C₈ cycloalkyl), C(=O)(optionally substituted C2-C8 heterocycloalkyl), C(=O)(optionally substituted C₆-C₁₀ aryl), C(=O)(optionally substituted C₂-C₁₀ heteroaryl), optionally substituted C1-C6 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted C2-C10 heteroaryl. 2. The compound of claim 1, wherein exactly one of R¹, R², R^3a, R^3b, R^4a, R^4b, R^4c, and R^4d is R⁵. 3. The compound of claim 1 or 2, wherein the compound of Formula (I) is a compound of Formula (Ia), or a salt, solvate, stereoisomer, or isotopologue thereof: , R² is selected from the substituted C1-C6 alkyl, optionally substituted C₃-C₈ cycloalkyl, optionally substituted C₂-C₈ heterocycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted heteroaryl; R^3a and R^3b are each independently selected from the group consisting of H, and optionally substituted C1-C6 alkyl; R^4a, R^4b, R^4c, and R^4d are each independently selected from the group consisting of H, - 103 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) halogen, CN, NO2, optionally substituted C1-C6 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C₂-C₈ heterocycloalkyl, optionally substituted C₆-C₁₀ aryl, optionally substituted C2-C10 heteroaryl, OR^A, N(R^A)(R^B), C(=O)N(R^A)(R^B), C(=O)R^A, C(=O)OR^A, OC(=O)R^A, OC(=O)OR^A, SR^A, S(=O)R^A, S(=O)₂R^A, N(R^A)S(=O)₂R^B, N(R^A)C(=O)R^B, and S(=O)2N(R^A)(R^B); R^6a and R^6b are each independently selected from the group consisting of -(optionally substituted C1-C6 alkylenyl)-C(=O)OR^7a, -(optionally substituted C1-C6 alkylenyl)- C(=O)N(R^7a)(R^7b), -(optionally substituted C₁-C₆ alkylenyl)-C(=O)R^7a, -(optionally substituted C1-C6 alkylenyl)-(R^7a), -C(=O)OR^7a, -C(=O)N(R^7a)(R^7b), -C(=O)R^7a, and R^7a; each occurrence of R^7a and R^7b is independently selected from the group consisting of optionally substituted C1-C28 alkyl, optionally substituted C2-C28 heteroalkyl, optionally substituted C₃-C₈ cycloalkyl, optionally substituted C₂-C₈ heterocycloalkyl, optionally substituted C2-C28 alkenyl, and optionally substituted C2-C28 alkynyl; L is selected from the group consisting of a bond, optionally substituted C₁-C₆ alkylenyl, optionally substituted C2-C6 alkenylenyl, optionally substituted C2-C6 heteroalkylenyl, optionally substituted C3-C8 cycloalkylenyl, optionally substituted C2-C8 heterocycloalkylenyl, optionally substituted C7-C12 aralkylenyl, optionally substituted C5-C12 heteroaralkylenyl, optionally substituted C6-C10 arylenyl, and optionally substituted C2-C10 heteroarylenyl; and each occurrence of R^A and R^B is independently selected from the group consisting of H, C(=O)(optionally substituted C₁-C₆ alkyl), C(=O)(optionally substituted C₃-C₈ cycloalkyl), C(=O)(optionally substituted C2-C8 heterocycloalkyl), C(=O)(optionally substituted C₆-C₁₀ aryl), C(=O)(optionally substituted C₂-C₁₀ heteroaryl), optionally substituted C1-C6 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C6-C10 aryl, and optionally substituted C₂-C₁₀ heteroaryl. 4. The compound of any one of claims 1-3, wherein R² is n-butyl. 5. The compound of any one of claims 1-4, wherein R^3a and R^3b are each independently H. 6. The compound of any one of claims 1-5, wherein at least one of the following applies: (a) at least one of R^4a, R^4b, R^4c, and R^4d is H; (b) at least two of R^4a, R^4b, R^4c, and R^4d are H; - 104 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) (c) at least three of R^4a, R^4b, R^4c, and R^4d are H; (d) each of R^4a, R^4b, R^4c, and R^4d are H. 7. The compound of any one of claims 1-6, wherein L is optionally substituted C₇-C₁₂ aralkylenyl. 8. The compound of any one of claims 1-7, wherein L . 9. The compound of any one of claims 1-8, wherein R^6a and R^6b are each independently selected from the group consisting of R^7a and -CH2CH2C(=O)OR^7a. 10. The compound of any one of claims 1-9, wherein R^7a and R^7b are each independently selected from the group consisting of optionally substituted C₆-C₂₈ alkyl, optionally substituted C6-C28 alkenyl, and optionally substituted C6-C28 heteroalkyl. 11. The compound of any one of claims 1-10, wherein R^6a and R^6b are each independently selected from the group consisting of -CH2CH(OH)(optionally substituted C6-C28 alkyl), - CH₂CH(OH)(optionally substituted C₆-C₂₈ alkenyl), -CH₂CH(OH)(optionally substituted C₆- C28 heteroalkyl), -CH2CH2C(=O)O(optionally substituted C6-C28 alkyl), - CH₂CH₂C(=O)O(optionally substituted C₆-C₂₈ alkenyl), and -CH₂CH₂C(=O)O(optionally substituted C6-C28 heteroalkyl). 12. The compound of any one of claims 1-11, wherein R^6a and R^6b are each independently selected from the group consisting of -CH₂CH(OH)(CH₂)₉CH₃ and - CH2CH2C(=O)O(CH2)11CH3. 13. The compound of any one of claims 1-12, which is selected from the group consisting of: - 105 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) . 14. (a) at least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent; (b) at least one ionizable lipid; (c) at least one helper lipid; (d) cholesterol; and (e) at least one polymer conjugated lipid. 15. The LNP of claim 14, wherein the hydrocarbyl substituted TLR agonist is a compound of Formula (II): A(B)n (II), wherein: A is a toll-like receptor (TLR) agonist; each occurrence of B is independently selected from the group consisting of R^6a and ; each occurrence of R^6a and R^6b is independently selected from the group consisting of -(optionally substituted C1-C6 alkylenyl)-C(=O)OR^7a, -(optionally substituted C1-C6 alkylenyl)-C(=O)N(R^7a)(R^7b), -(optionally substituted C₁-C₆ alkylenyl)-C(=O)R^7a, - (optionally substituted C1-C6 alkylenyl)-(R^7a), -C(=O)OR^7a, -C(=O)N(R^7a)(R^7b), -C(=O)R^7a, and R^7a, each occurrence of R^7a and R^7b is independently selected from the group consisting of - 106 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) optionally substituted C1-C28 alkyl, optionally substituted C2-C28 heteroalkyl, optionally substituted C₃-C₈ cycloalkyl, optionally substituted C₂-C₈ heterocycloalkyl, optionally substituted C2-C28 alkenyl, and optionally substituted C2-C28 alkynyl; L is bond, optionally substituted C₁-C₆ alkylenyl, optionally substituted C₂-C₆ alkenylenyl, optionally substituted C2-C6 heteroalkylenyl, optionally substituted C3-C8 cycloalkylenyl, optionally substituted C₂-C₈ heterocycloalkylenyl, optionally substituted C₇- C12 aralkylenyl, optionally substituted C5-C12 heteroaralkylenyl, optionally substituted C6-C10 arylenyl, and optionally substituted C₂-C₁₀ heteroarylenyl; and n is an integer selected from the group consisting of 1, 2, 3, and 4. 16. The LNP of claim 14 or 15, wherein the TLR agonist is selected from the group consisting of 1-(4-(aminomethyl)benzyl)-2-butyl-1H-imidazo[4,5-c]quinolin-4-amine (TLR7/8 agonist 1), 1-isobutyl-1H-imidazo[4,5-c]quinolin-4-amine (imiquimod), 1-(4- amino-2-(ethoxymethyl)-1H-imidazo[4,5-c]quinolin-1-yl)-2-methylpropan-2-ol (resiquimod), and 1-(4-amino-2-((ethylamino)methyl)-1H-imidazo[4,5-c]quinolin-1-yl)-2-methylpropan-2- ol (gardiquimod). 17. The LNP of claim 14, wherein the least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent is the compound of Formula (I) of any one of claims 1-13. 18. The LNP of any one of claims 14-17, wherein the at least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent is selected from the group consisting of: . - 107 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) 19. The LNP of any one of claims 14-18, wherein the ionizable lipid is at least one selected from the group consisting of: , R^8a, R^8b, R^8c, R^8d, R^8e, and R^8f are each independently selected from the group consisting of -(optionally substituted C1-C6 alkylenyl)-C(=O)OR^9a, -(optionally substituted C₁-C₆ alkylenyl)-C(=O)N(R^9a)(R^9b), -(optionally substituted C₁-C₆ alkylenyl)-C(=O)R^9a, - (optionally substituted C1-C6 alkylenyl)-(R^9a), -C(=O)OR^9a, -C(=O)N(R^9a)(R^9b), -C(=O)R^9a, and R^9a; and each occurrence of R^9a and R^9b is independently selected from the group consisting of optionally substituted C₁-C₂₈ alkyl, optionally substituted C₂-C₂₈ heteroalkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C₂-C₂₈ alkenyl, and optionally substituted C₂-C₂₈ alkynyl. 20. The LNP of claim 19, wherein R^8a, R^8b, R^8c, R^8d, R^8e, and R^8f are each independently selected from the group consisting of R^9a and -CH2CH2C(=O)OR^9a. 21. The LNP of claim 19 or 20, wherein R^9a and R^9b are each independently selected from the group consisting of optionally substituted C1-C28 alkyl, optionally substituted C1-C28 alkenyl, and optionally substituted C₁-C₂₈ heteroalkyl. 22. The LNP of any one of claims 19-21, wherein R^7a and R^7b are each independently selected from the group consisting of -CH2CH(OH)(optionally substituted C1-C28 alkyl), - - 108 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) CH2CH(OH)(optionally substituted C1-C28 alkenyl), -CH2CH(OH)(optionally substituted C2- C₂₈ heteroalkyl), -CH₂CH₂C(=O)O(optionally substituted C₁-C₂₈ alkyl), - CH2CH2C(=O)O(optionally substituted C1-C28 alkenyl), and -CH2CH2C(=O)O(optionally substituted C₂-C₂₈ heteroalkyl). 23. The LNP of any one of claims 19-22, wherein R^7a and R^7b are each independently selected from the group consisting of -CH2CH(OH)(CH2)9CH3 and - CH₂CH₂C(=O)O(CH₂)₁₁CH₃. 24. The LNP of any one of claims 14-23, wherein the at least one ionizable lipid comprises: . 25. The LNP of any one of claims 14-24, wherein the at least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent comprises about 1 mol% to about 17.5 mol% of the LNP. 26. The LNP of any one of claims 14-25, wherein the at least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent comprises about 1, 2.5, 5, 10, or about 17.5 mol% of the LNP. 27. The LNP of any one of claims 14-26, wherein the at least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent comprises about 5 mol% of the LNP. 28. The LNP of any one of claims 14-27, wherein the at least one ionizable lipid comprises about 10 mol% to about 60 mol% of the LNP. 29. The LNP of any one of claims 14-28, wherein the at least one ionizable lipid comprises about 34, 32.5, 30, 25, or about 17.5 mol% of the LNP. - 109 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) 30. The LNP of any one of claims 14-29, wherein the at least one ionizable lipid comprises about 30 mol% of the LNP. 31. The LNP of any one of claims 14-30, wherein the at least one helper lipid comprises at least one selected from the group consisting of dioleoylphosphatidylethanolamine (DOPE) and distearoylphosphatidylcholine (DSPC). 32. The LNP of any one of claims 14-31, wherein the at least one helper lipid comprises about 1 mol% to about 50 mol% of the LNP. 33. The LNP of any one of claims 14-32, wherein the at least one helper lipid comprises about 16 mol% of the LNP. 34. The LNP of any one of claims 14-33, wherein the cholesterol comprises about 5 mol% to about 60 mol% of the LNP. 35. The LNP of any one of claims 14-34, wherein the cholesterol comprises about 46.5 mol% of the LNP. 36. The LNP of any one of claims 14-35, wherein the at least one polymer conjugated lipid comprises 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (DMG-PEG 2000). 37. The LNP of any one of claims 14-36, wherein the at least one polymer conjugated lipid comprises about 0.1 mol% to about 20 mol% of the LNP. 38. The LNP of any one of claims 14-37, wherein the at least one polymer conjugated lipid comprises about 2.5 mol% of the LNP. 39. The LNP of any one of claims 14-38, wherein the LNP has a molar ratio of (a) : (b) : (c) : (d) : (e) of about 5 : 30 : 16 : 46.5 : 2.5. 40. The LNP of any one of claims 14-39, wherein the LNP further comprises at least one - 110 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) cargo molecule. 41. The LNP of claim 40, wherein the cargo is at least one selected from the group consisting of a nucleic acid, small molecule, protein, therapeutic agent, antibody, and any combinations thereof. 42. The LNP of claim 40 or 41, wherein the cargo is a nucleic acid. 43. The LNP of claim 41 or 42, wherein the nucleic acid is DNA or RNA. 44. The LNP of any one of claims 41-43, wherein the nucleic acid is selected from the group consisting of mRNA, cDNA, pDNA, microRNA, siRNA, modified RNA, antagomir, antisense molecule, and any combinations thereof. 45. The LNP of any one of claims 40-44, wherein the cargo is at least partially encapsulated in the LNP. 46. The LNP of any one of claims 40-45, wherein the cargo is mRNA. 47. The LNP of claim 46, wherein the LNP has a weight ratio of total lipidoid (i.e., hydrocarbyl substituted toll-like receptor agonist and ionizable lipid) to mRNA ranging from about 5:1 to about 20:1 (i.e., weight ratio of (a) + (b) : mRNA). 48. The LNP of claim 46 or 47, wherein the LNP has a weight ratio of total lipidoid (i.e., hydrocarbyl substituted toll-like receptor agonist and ionizable lipid) to mRNA of about 10:1 (i.e., weight ratio of (a) + (b) : mRNA). 49. The LNP of any one of claims 44-48, wherein the mRNA encodes SARS-CoV-2, an immunogenic fragment thereof (e.g., spike protein), or a modified derivative thereof. 50. A pharmaceutical composition comprising the lipid nanoparticle (LNP) of any one of claims 14-49 and a pharmaceutically acceptable carrier. - 111 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) 51. A method of generating an innate immune response in a subject, the method comprising administering to the subject the lipid nanoparticle (LNP) of any one of claims 14- 49 or the pharmaceutical composition of claim 50. 52. A method of treating, preventing, and/or ameliorating an infection, disease, or disorder in a subject, the method comprising administering to the subject a lipid nanoparticle (LNP) comprising: (a) at least one toll-like receptor (TLR) agonist substituted with at least one hydrocarbyl substituent; (b) at least one ionizable lipid; (c) at least one helper lipid; (d) cholesterol; (e) at least one polymer conjugated lipid; and (f) at least one cargo molecule. 53. The method of claim 52, wherein the cargo is a nucleic acid. 54. The method of claim 52 or 53, wherein the nucleic acid is DNA or RNA. 55. The method of any one of claims 52-54, wherein the nucleic acid is selected from the group consisting of mRNA, cDNA, pDNA, microRNA, siRNA, modified RNA, antagomir, antisense molecule, and any combinations thereof. 56. The method of any one of claims 52-55, wherein the cargo is at least partially encapsulated in the LNP. 57. The method of any one of claims 52-56, wherein the cargo is mRNA. 58. The method of claim 57, wherein the mRNA encodes SARS-CoV-2, an immunogenic fragment thereof (e.g., spike protein), or a modified derivative thereof. 59. The method of claim 58, wherein the infection, disease, or disorder is a SARS-CoV-2 - 112 - 52436892.1 Attorney Docket No.046483-7434WO1(03633) infection. 60. The method of any one of claims 52-59, wherein an innate immune response is promoted in the subject. 61. The method of any one of claims 51-60, wherein the subject is a mammal. 62. The method of claim 61, wherein the mammal is a human. - 113 - 52436892.1