Galnac lipid compounds for use in lipid nanoparticles

Abstract

Compounds having the structure of Formula (I): or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof; wherein R1, R2, R3, R4, R5, L1, L2, a and z are as defined herein, are disclosed. Use of these compounds as a component of lipid nanoparticle formulations for delivery of a therapeutic agent, compositions comprising the compounds, and methods for their use and preparation are also provided.

Description

[0001] GALNAC LIPID COMPOUNDS FOR USE IN LIPID NANOPARTICLES Technical Field The present disclosure generally relates to novel GalNAc-lipids that can be used in combination with other lipid components, such as cationic lipids, neutral lipids, cholesterol, and polymer conjugated lipids, to form lipid nanoparticles, to facilitate the intracellular delivery of therapeutic nucleic acids (e.g., oligonucleotides, messenger RNA) both in vitro and in vivo. Description of Related Art There are many challenges associated with the delivery of nucleic acids to elicit a desired response in a biological system. Nucleic acid-based therapeutics have enormous potential but there remains a need for more effective delivery of nucleic acids to appropriate sites within a cell, tissue, or an organism to realize this potential. Therapeutic nucleic acids include, e.g., messenger RNA (mRNA), antisense oligonucleotides, ribozymes, DNAzymes, plasmids, immune stimulating nucleic acids, antagomir, antimir, mimic, supermir, and aptamers. Some nucleic acids, such as mRNA or plasmids, can be used to induce expression of specific cellular products as would be useful in the treatment of, for example, diseases related to a deficiency of a protein or enzyme. The therapeutic applications of translatable nucleotide delivery are extremely broad as constructs can be synthesized to produce any chosen protein sequence, whether indigenous to the system. The expression products of the nucleic acid can augment existing levels of protein, replace missing or non-functional versions of a protein, or introduce a new protein and associated functionality in a cell or organism. BRIEF SUMMARY Nucleic acids, such as miRNA inhibitors, can be used to modulate the expression of specific cellular products that are regulated by miRNA offering therapeutic potential in the treatment of, for example, diseases associated with protein or enzyme deficiencies. The therapeutic applications of miRNA inhibition are extremely broad as constructs can be synthesized to inhibit one or more miRNA that would in turn regulate the expression of mRNA products. The inhibition of endogenous miRNA can augment its downstream target endogenous protein expression and restore proper function in a cell or organism to treat disease associated with a specific miRNA or a group of miRNA.

[0002] 1 Other nucleic acids can down-regulate intracellular levels of specific mRNA and, as a result, down-regulate the synthesis of the corresponding proteins through processes such as RNA interference (RNAi) or complementary binding of antisense RNA. The therapeutic applications of antisense oligonucleotide and RNAi are also extremely broad, since oligonucleotide constructs can be synthesized with any nucleotide sequence directed against a target mRNA. Targets may include mRNAs from normal cells, mRNAs associated with disease-states, such as cancer, and mRNAs of infectious agents, such as viruses. To date, antisense oligonucleotide constructs have shown the ability to specifically down-regulate target proteins through degradation of the cognate mRNA in both in vitro and in vivo models. In addition, antisense oligonucleotide constructs are currently being evaluated in clinical studies. However, two problems currently face using oligonucleotides in therapeutic contexts. First, free RNAs are susceptible to nuclease digestion in plasma. Second, free RNAs have limited ability to gain access to the intracellular compartment where the relevant translation machinery resides. Lipid nanoparticles formed from lipid components, such as cationic lipids, neutral lipids, cholesterol, PEG, PEGylated lipids, and oligonucleotides have been used to block degradation of the RNAs in plasma and facilitate the cellular uptake of the oligonucleotides. Lipid nanoparticles have demonstrated utility in hepatic delivery of oligonucleotides typically via low-density lipoprotein receptor-mediated endocytosis. However, there remains a need for improved lipid nanoparticles compositions for the delivery of oligonucleotides. In particular, for patients lacking sufficient low-density lipoprotein receptor activity, there remains a need for improved lipid nanoparticles for hepatic delivery of oligonucleotides. Preferably, these lipid nanoparticles would provide optimal drug to lipid ratios, protect the nucleic acid from degradation and clearance in serum, be suitable for intramuscular or systemic delivery, and provide intracellular delivery of the nucleic acid. In addition, these lipid- nucleic acid particles should be well-tolerated and provide an adequate therapeutic index, such that patient treatment at an effective dose of the nucleic acid is not associated with unacceptable toxicity and / or risk to the patient. The present disclosure provides these and related advantages. In brief, the present disclosure provides GalNAc-lipid compounds, including stereoisomers, pharmaceutically acceptable salts, stereoisomers, and tautomers thereof, which can be used alone or in combination with other lipid components such as cationic lipids, neutral lipids, charged lipids, steroids (including for example, all sterols) and / or their analogs, and / or polymer conjugated lipids to form lipid nanoparticles for the delivery of therapeutic agents. In some instances, the lipid nanoparticles are used to deliver nucleic acids such as antisense and / or messenger RNA. Methods for use of such lipid nanoparticles for treatment or prevention (e.g., vaccination) of various diseases or conditions, such as those caused by infectious entities and / or insufficiency of a protein, are also provided. Accordingly, in one embodiment is provided a compound having the structure of Formula (I): or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof; wherein R1, R2, R3, R4, R5, L1, L2, a, and z are as defined herein. Lipid nanoparticles comprising the compound of Formula (I) and a therapeutic agent, as well as compositions comprising the compound of Formula (I) and a therapeutic agent are provided in other embodiments. In some embodiments, the lipid nanoparticles and compositions further comprise one or more components selected from neutral lipids, charged lipids, steroids, and polymer conjugated lipids. Also disclosed are pharmaceutical compositions comprising the lipid nanoparticles or compositions comprising the compound of Formula (I). In other embodiments, the present disclosure provides a method for administering a therapeutic agent to a patient in need thereof, the method comprising preparing a composition of lipid nanoparticles comprising GalNAc-lipids (e.g., compounds of Formula (I)) and a therapeutic agent and delivering the composition to the patient. Such methods are useful for inducing expression of a protein in a subject, for example for expressing an antigen for purposes of vaccination or a gene editing protein. These and other aspects of the disclosure will be apparent upon reference to the following detailed description. BRIEF DESCRIPTION OF SEVERAL VIEWS OF THE DRAWINGS Figure 1 shows that LNPs comprising GalNAc lipids of the disclosure have improved activity compared to LNPs without the disclosed GalNAc and LNPs comprising a prior art GalNAc lipid in both ApoE & LDLR knock out mice at both 24 hours (1A) and 48 hours (1B). Figure 2 provides IgG expression level for non-human primates treated with LNP comprising a cationic lipid of formula III (Figure 2A; 0.5 mg / kg and 1.5 mg / kg) or formula IV (Figure 2B; 0.5 mg / kg and 1.5 mg / kg), with and without a GalNAc pegylated lipid of the present disclosure. Figure 3 shows serum IgG expression following administration of LNPII (LNP comprising a cationic lipid of Formula II) at 0.3 mg / kg (with and without compound 1-1 and compound 3) in wild type and LDLR knockout mice. For additional information and explanation, see Example 3. Figure 4 shows serum IgG expression following administration of LNPIII (LNP comprising a cationic lipid of Formula III) at 0.5 mg / kg (with and without compound 1-1 and compound 3) in wild type and LDLR knockout mice. For additional information and explanation, see Example 3. Figure 5 shows serum IgG expression at a dose of 0.15 mg / kg (Figure 5A), 0.3 mg / kg (Figure 5B), and 0.6 mg / kg (Figure 5C) for LNPs as indicated (i.e., from left to right – LNPIV, LNPIV with compound 1-1, and LNPIV with compound 3). For additional information and explanation, see Example 4. Figure 6 shows peak plasma IgG concentration following the administration of LNPII with and without a GalNAc lipid at doses of 0.5 mg / kg and 1.5 mg / kg. For additional information and explanation, see Example 5. Figure 7 shows area under the curve (AUC; Figure 7A) and Cmax (Figure 7B) plasma IgG concentrations for LNPIV dosed at 1.5 mg / kg. For additional information and explanation, see Example 6. DETAILED DESCRIPTION In the following description, certain specific details are set forth to provide a thorough understanding of various embodiments of the disclosure. However, one skilled in the art will understand that the embodiments of the disclosure may be practiced without these details. The present disclosure is based, in part, upon the discovery of novel GalNAc lipids that provide advantages when used in lipid nanoparticles (LNPs) for the in vivo delivery of an active or therapeutic agent such as a nucleic acid into a cell of an animal, such as a mammal (e.g., human). In some embodiments, the present disclosure provides nucleic acid-lipid nanoparticle compositions comprising one or more of the novel GalNAc lipids described herein that provide increased activity of the nucleic acid and improved tolerability of the compositions in vivo, resulting in a significant increase in the therapeutic index as compared to nucleic acid-lipid nanoparticle compositions previously described. In some embodiments, the present disclosure provides novel GalNAc lipids that enable the formulation of improved compositions for the in vitro and in vivo delivery of mRNA and / or other oligonucleotides. In some embodiments, these improved lipid nanoparticle compositions are useful for expression of protein encoded by mRNA. In other embodiments, these improved LNPs compositions are useful for upregulation of endogenous protein expression by delivering miRNA inhibitors targeting one specific miRNA or a group of miRNA regulating one target mRNA or several mRNA. In other embodiments, these improved lipid nanoparticle compositions are useful for down-regulating (e.g., silencing) the protein levels and / or mRNA levels of target genes. In some other embodiments, the LNPs are also useful for delivery of mRNA and plasmids for expression of transgenes. In yet other embodiments, the lipid nanoparticle compositions are useful for inducing a pharmacological effect resulting from expression of a protein, e.g., increased production of red blood cells through the delivery of a suitable erythropoietin mRNA, or protection against infection through delivery of mRNA encoding for a suitable antibody. The LNPs and compositions of the present disclosure may be used for a variety of purposes, including the delivery of encapsulated or associated (e.g., complexed) therapeutic agents such as nucleic acids to cells, both in vitro and in vivo. Accordingly, embodiments of the present disclosure provide methods of treating or preventing diseases or disorders in a subject in need thereof by contacting the subject with a lipid nanoparticle that encapsulates or is associated with a suitable therapeutic agent, wherein the lipid nanoparticle comprises one or more of the novel GalNAc lipids described herein. As described herein, embodiments of the LNPs of the present disclosure are particularly useful for the delivery of nucleic acids, including, e.g., mRNA, antisense oligonucleotide, plasmid DNA, microRNA (miRNA), miRNA inhibitors (antagomirs / antimirs), messenger-RNA- interfering complementary RNA (micRNA), DNA, multivalent RNA, dicer substrate RNA, complementary DNA (cDNA), etc. Therefore, the LNPs and compositions of the present disclosure may be used to induce expression of a desired protein both in vitro and in vivo by contacting cells with a lipid nanoparticle comprising one or more novel GalNAc lipids described herein, wherein the lipid nanoparticle encapsulates or is associated with a nucleic acid that is expressed to produce the desired protein (e.g., a messenger RNA or plasmid encoding the desired protein). Alternatively, the LNPs and compositions of the present disclosure may be used to decrease the expression of target genes and proteins both in vitro and in vivo by contacting cells with a lipid nanoparticle comprising one or more novel GalNAc lipids described herein, wherein the lipid nanoparticle encapsulates or is associated with a nucleic acid that reduces target gene expression (e.g., an antisense oligonucleotide or small interfering RNA (siRNA)). The LNPs and compositions of the present disclosure may also be used for co-delivery of different nucleic acids (e.g., mRNA and plasmid DNA) separately or in combination, such as may be useful to provide an effect requiring colocalization of different nucleic acids (e.g., mRNA encoding for a suitable gene modifying enzyme and DNA segment(s) for incorporation into the host genome). Nucleic acids for use with this disclosure may be prepared according to any available technique. For mRNA, the primary methodology of preparation is, but not limited to, enzymatic synthesis (also termed in vitro transcription) which currently represents the most efficient method to produce long sequence-specific mRNA. In vitro transcription describes a process of template-directed synthesis of RNA molecules from an engineered DNA template comprised of an upstream bacteriophage promoter sequence (e.g., including but not limited to that from the T7, T3, and SP6 coliphage) linked to a downstream sequence encoding the gene of interest. Template DNA can be prepared for in vitro transcription from a number of sources with appropriate techniques which are well known in the art including, but not limited to, plasmid DNA and polymerase chain reaction amplification (see Linpinsel, J.L. and Conn, G.L., General protocols for preparation of plasmid DNA template and Bowman, J.C., Azizi, B., Lenz, T.K., Ray, P., and Williams, L.D. in RNA in vitro transcription and RNA purification by denaturing PAGE in Recombinant and in vitro RNA syntheses Methods v.941 Conn G.L. (ed), New York, N.Y. Humana Press, 2012). Transcription of the RNA occurs in vitro using the linearized DNA template in the presence of the corresponding RNA polymerase and adenosine, guanosine, uridine, and cytidine ribonucleoside triphosphates (rNTPs) under conditions that support polymerase activity while minimizing potential degradation of the resultant mRNA transcripts. In vitro transcription can be performed using a variety of commercially available kits including but not limited to RiboMax Large Scale RNA Production System (Promega), MegaScript Transcription kits (Life Technologies) as well as with commercially available reagents including RNA polymerases and rNTPs. The methodology for in vitro transcription of mRNA is well known in the art. (see, e.g., Losick, R., 1972, In vitro transcription, Ann Rev Biochem v.41409-46; Kamakaka, R. T. and Kraus, W. L.2001. In Vitro Transcription. Current Protocols in Cell Biology.2:11.6:11.6.1– 11.6.17; Beckert, B. And Masquida, B., (2010) Synthesis of RNA by In Vitro Transcription in RNA in Methods in Molecular Biology v.703 (Neilson, H. Ed), New York, N.Y. Humana Press, 2010; Brunelle, J.L. and Green, R., 2013, Chapter Five – In vitro transcription from plasmid or PCR-amplified DNA, Methods in Enzymology v.530, 101-114; all of which are incorporated herein by reference). The desired in vitro transcribed mRNA is then purified from the undesired components of the transcription or associated reactions (including unincorporated rNTPs, protein enzyme, salts, short RNA oligos etc.). Techniques for the isolation of the mRNA transcripts are well known in the art. Well known procedures include phenol / chloroform extraction or precipitation with either alcohol (ethanol, isopropanol) in the presence of monovalent cations or lithium chloride. Additional, non-limiting examples of purification procedures which can be used include size exclusion chromatography (Lukavsky, P.J. and Puglisi, J.D., 2004, Large-scale preparation and purification of polyacrylamide-free RNA oligonucleotides, RNA v.10, 889-893), silica-based affinity chromatography and polyacrylamide gel electrophoresis (Bowman, J.C., Azizi, B., Lenz, T.K., Ray, P., and Williams, L.D. in RNA in vitro transcription and RNA purification by denaturing PAGE in Recombinant and in vitro RNA syntheses Methods v.941 Conn G.L. (ed), New York, N.Y. Humana Press, 2012 ). Purification can be performed using a variety of commercially available kits including but not limited to SV Total Isolation System (Promega) and In Vitro Transcription Cleanup and Concentration Kit (Norgen Biotek). Furthermore, while reverse transcription can yield large quantities of mRNA, the products can contain several aberrant RNA impurities associated with undesired polymerase activity which may need to be removed from the full-length mRNA preparation. These include short RNAs that result from abortive transcription initiation as well as double-stranded RNA (dsRNA) generated by RNA-dependent RNA polymerase activity, RNA-primed transcription from RNA templates and self-complementary 3'-extension. It has been demonstrated that these contaminants with dsRNA structures can lead to undesired immunostimulatory activity through interaction with various innate immune sensors in eukaryotic cells that function to recognize specific nucleic acid structures and induce potent immune responses. This in turn can dramatically reduce mRNA translation since protein synthesis is reduced during the innate cellular immune response. Therefore, additional techniques to remove these dsRNA contaminants have been developed and are known in the art including but not limited to scaleable HPLC purification (see, e.g., Kariko, K., Muramatsu, H., Ludwig, J. And Weissman, D., 2011, Generating the optimal mRNA for therapy: HPLC purification eliminates immune activation and improves translation of nucleoside-modified, protein-encoding mRNA, Nucl Acid Res, v.39 e142; Weissman, D., Pardi, N., Muramatsu, H., and Kariko, K., HPLC Purification of in vitro transcribed long RNA in Synthetic Messenger RNA and Cell Metabolism Modulation in Methods in Molecular Biology v.969 (Rabinovich, P.H. Ed), 2013). HPLC purified mRNA has been reported to be translated at much greater levels, particularly in primary cells and in vivo. A significant variety of modifications have been described in the art which are used to alter specific properties of in vitro transcribed mRNA and improve its utility. These include but are not limited to modifications to the 5'- and 3'-termini of the mRNA. Endogenous eukaryotic mRNA typically contains a cap structure on the 5′-end of a mature molecule which plays an important role in mediating binding of the mRNA Cap Binding Protein (CBP), which is in turn responsible for enhancing mRNA stability in the cell and efficiency of mRNA translation. Therefore, the highest levels of protein expression are achieved with capped mRNA transcripts. The 5′-cap contains a 5′-5′-triphosphate linkage between the 5′-most nucleotide and guanine nucleotide. The conjugated guanine nucleotide is methylated at the N7 position. Additional modifications include methylation of the ultimate and penultimate most 5′-nucleotides on the 2′- hydroxyl group. Multiple distinct cap structures can be used to generate the 5′-cap of in vitro transcribed synthetic mRNA.5'-capping of synthetic mRNA can be performed co-transcriptionally with chemical cap analogs (i.e., capping during in vitro transcription). For example, the Anti-Reverse Cap Analog (ARCA) cap contains a 5′-5′-triphosphate guanine-guanine linkage where one guanine contains an N7 methyl group as well as a 3′-O-methyl group. However, up to 20% of transcripts remain uncapped during this co-transcriptional process and the synthetic cap analog is not identical to the 5′-cap structure of an authentic cellular mRNA, potentially reducing translatability and cellular stability. Alternatively, synthetic mRNA molecules may also be enzymatically capped post-transcriptionally. These may generate a more authentic 5′-cap structure that more closely mimics, either structurally or functionally, the endogenous 5'-cap which have enhanced binding of cap binding proteins, increased half-life, reduced susceptibility to 5′ endonucleases and / or reduced 5′ de-capping. Numerous synthetic 5'-cap analogs have been developed and are known in the art to enhance mRNA stability and translatability (see, e.g., Grudzien-Nogalska, E., Kowalska, J., Su, W., Kuhn, A.N., Slepenkov, S.V., Darynkiewicz, E., Sahin, U., Jemielity, J., and Rhoads, R.E., Synthetic mRNAs with superior translation and stability properties in Synthetic Messenger RNA and Cell Metabolism Modulation in Methods in Molecular Biology v.969 (Rabinovich, P.H. Ed), 2013). On the 3'-terminus, a long chain of adenine nucleotides (poly-A tail) is normally added to mRNA molecules during RNA processing. Immediately after transcription, the 3′ end of the transcript is cleaved to free a 3′ hydroxyl to which poly-A polymerase adds a chain of adenine nucleotides to the RNA in a process called polyadenylation. The poly-A tail has been extensively shown to enhance both translational efficiency and stability of mRNA (see Bernstein, P. and Ross, J., 1989, Poly (A), poly (A) binding protein and the regulation of mRNA stability, Trends Bio Sci v.14373-377; Guhaniyogi, J. And Brewer, G., 2001, Regulation of mRNA stability in mammalian cells, Gene, v.265, 11-23; Dreyfus, M. And Regnier, P., 2002, The poly (A) tail of mRNAs: Bodyguard in eukaryotes, scavenger in bacteria, Cell, v.111, 611-613). Poly (A) tailing of in vitro transcribed mRNA can be achieved using various approaches including, but not limited to, cloning of a poly (T) tract into the DNA template or by post- transcriptional addition using Poly (A) polymerase. The first case allows in vitro transcription of mRNA with poly (A) tails of defined length, depending on the size of the poly (T) tract, but requires additional manipulation of the template. The latter case involves the enzymatic addition of a poly (A) tail to in vitro transcribed mRNA using poly (A) polymerase which catalyzes the incorporation of adenine residues onto the 3'-termini of RNA, requiring no additional manipulation of the DNA template, but results in mRNA with poly(A) tails of heterogenous length.5'-capping and 3'-poly (A) tailing can be performed using a variety of commercially available kits including, but not limited to Poly (A) Polymerase Tailing kit (EpiCenter), mMESSAGE mMACHINE T7 Ultra kit and Poly (A) Tailing kit (Life Technologies) as well as with commercially available reagents, various ARCA caps, Poly (A) polymerase, etc. In addition to 5'-cap and 3'-poly adenylation, other modifications of the in vitro transcripts have been reported to provide benefits as related to efficiency of translation and stability. It is well known in the art that pathogenic DNA and RNA can be recognized by a variety of sensors within eukaryotes and trigger potent innate immune responses. The ability to discriminate between pathogenic and self-DNA and RNA has been shown to be based, at least in part, on structure and nucleoside modifications since most nucleic acids from natural sources contain modified nucleosides. In contrast, in vitro synthesized RNA lacks these modifications, thus rendering it immunostimulatory which in turn can inhibit effective mRNA translation as outlined above. The introduction of modified nucleosides into in vitro transcribed mRNA can be used to prevent recognition and activation of RNA sensors, thus mitigating this undesired immunostimulatory activity and enhancing translation capacity (see, e.g., Kariko, K. And Weissman, D.2007, Naturally occurring nucleoside modifications suppress the immunostimulatory activity of RNA: implication for therapeutic RNA development, Curr Opin Drug Discov Devel, v.10523-532; Pardi, N., Muramatsu, H., Weissman, D., Kariko, K., In vitro transcription of long RNA containing modified nucleosides in Synthetic Messenger RNA and Cell Metabolism Modulation in Methods in Molecular Biology v.969 (Rabinovich, P.H. Ed), 2013); Kariko, K., Muramatsu, H., Welsh, F.A., Ludwig, J., Kato, H., Akira, S., Weissman, D., 2008, Incorporation of Pseudouridine Into mRNA Yields Superior Nonimmunogenic Vector With Increased Translational Capacity and Biological Stability, Mol Ther v.16, 1833-1840. The modified nucleosides and nucleotides used in the synthesis of modified RNAs can be prepared, monitored and utilized using general methods and procedures known in the art. A large variety of nucleoside modifications are available that may be incorporated alone or in combination with other modified nucleosides to some extent into the in vitro transcribed mRNA (see, e.g., US Publication No.2012 / 0251618). In vitro synthesis of nucleoside-modified mRNA has been reported to have reduced ability to activate immune sensors with a concomitant enhanced translational capacity. Other components of mRNA which can be modified to provide benefit in terms of translatability and stability include the 5′- and 3'-untranslated regions (UTR). Optimization of the UTRs (favorable 5'- and 3'-UTRs can be obtained from cellular or viral RNAs), either both or independently, have been shown to increase mRNA stability and translational efficiency of in vitro transcribed mRNA (see, e.g., Pardi, N., Muramatsu, H., Weissman, D., Kariko, K., In vitro transcription of long RNA containing modified nucleosides in Synthetic Messenger RNA and Cell Metabolism Modulation in Methods in Molecular Biology v.969 (Rabinovich, P.H. Ed), 2013). In addition to mRNA, other nucleic acid payloads may be used for this disclosure. For oligonucleotides, methods of preparation include but are not limited to chemical synthesis and enzymatic, chemical cleavage of a longer precursor, in vitro transcription as described above, etc. Methods of synthesizing DNA and RNA nucleotides are widely used and well known in the art (see, e.g., Gait, M. J. (ed.)Oligonucleotide synthesis: a practical approach, Oxford [Oxfordshire], Washington, D.C.: IRL Press, 1984; and Herdewijn, P. (ed.) Oligonucleotide synthesis: methods and applications, Methods in Molecular Biology, v.288 (Clifton, N.J.) Totowa, N.J.: Humana Press, 2005; both of which are incorporated herein by reference). For plasmid DNA, preparation for use with this disclosure commonly utilizes but is not limited to expansion and isolation of the plasmid DNA in vitro in a liquid culture of bacteria containing the plasmid of interest. The presence of a gene in the plasmid of interest that encodes resistance to a particular antibiotic (penicillin, kanamycin, etc.) allows those bacteria containing the plasmid of interest to selective grow in antibiotic-containing cultures. Methods of isolating plasmid DNA are widely used and well known in the art (see, e.g., Heilig, J., Elbing, K. L. and Brent, R (2001) Large-Scale Preparation of Plasmid DNA. Current Protocols in Molecular Biology.41:II:1.7:1.7.1–1.7.16; Rozkov, A., Larsson, B., Gillström, S., Björnestedt, R. and Schmidt, S. R. (2008), Large-scale production of endotoxin-free plasmids for transient expression in mammalian cell culture. Biotechnol. Bioeng., 99: 557–566; and US Patent No. 6,197,553). Plasmid isolation can be performed using a variety of commercially available kits including, but not limited to Plasmid Plus (Qiagen), GenJET plasmid MaxiPrep (Thermo) and PureYield MaxiPrep (Promega) kits as well as with commercially available reagents. Various exemplary embodiments of the GalNAc lipids of the present disclosure, LNPs and compositions comprising the same, and their use to deliver active or therapeutic agents such as nucleic acids to modulate gene and protein expression, are described in further detail below. As used herein, the following terms have the meanings ascribed to them unless specified otherwise. Unless the context requires otherwise, throughout the present specification and claims, the word “comprise” and variations thereof, such as, “comprises” and “comprising” are to be construed in an open and inclusive sense, that is, as “including, but not limited to”. Reference throughout this specification to “one embodiment” or “an embodiment” means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the present disclosure. Thus, the appearances of the phrases “in one embodiment” or “in an embodiment” in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the features, structures, or characteristics may be combined in any suitable manner in one or more embodiments. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which this disclosure belongs. As used in the specification and claims, the singular form “a”, “an” and “the” include plural references unless the context clearly dictates otherwise. The phrase “induce expression of a desired protein” refers to the ability of a nucleic acid to increase expression of the desired protein. To examine the extent of protein expression, a test sample (e.g., a sample of cells in culture expressing the desired protein) or a test mammal (e.g., a mammal such as a human or an animal model such as a rodent (e.g., mouse) or a non-human primate (e.g., monkey) model) is contacted with a nucleic acid (e.g., nucleic acid in combination with a lipid of the present disclosure). Expression of the desired protein in the test sample or test animal is compared to expression of the desired protein in a control sample (e.g., a sample of cells in culture expressing the desired protein) or a control mammal (e.g., a mammal such as a human or an animal model such as a rodent (e.g., mouse) or non-human primate (e.g., monkey) model) that is not contacted with or administered the nucleic acid. When the desired protein is present in a control sample or a control mammal, the expression of a desired protein in a control sample or a control mammal may be assigned a value of 1.0. In some embodiments, inducing expression of a desired protein is achieved when the ratio of desired protein expression in the test sample or the test mammal to the level of desired protein expression in the control sample or the control mammal is greater than 1, for example, about 1.1, 1.5, 2.0.5.0 or 10.0. When a desired protein is not present in a control sample or a control mammal, inducing expression of a desired protein is achieved when any measurable level of the desired protein in the test sample or the test mammal is detected. One of ordinary skill in the art will understand appropriate assays to determine the level of protein expression in a sample, for example dot blots, northern blots, in situ hybridization, ELISA, immunoprecipitation, enzyme function, and phenotypic assays, or assays based on reporter proteins that can produce fluorescence or luminescence under appropriate conditions. 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, RNA, and hybrids thereof. DNA may be in the form of antisense molecules, plasmid DNA, cDNA, PCR products, or vectors. RNA may be in the form of small hairpin RNA (shRNA), messenger RNA (mRNA), antisense RNA, miRNA, micRNA, multivalent RNA, dicer substrate RNA or 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, single nucleotide polymorphisms, 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)). “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 alkylhalides. The term “gene” refers to a nucleic acid (e.g., DNA or RNA) sequence that comprises partial length or entire length coding sequences necessary to produce a polypeptide or precursor polypeptide. The term “lipid” refers to a group of organic compounds that include, but are not limited to, esters of fatty acids and are generally characterized by being poorly soluble in water, but soluble in many organic solvents. A “GalNAc-lipid” refers to a lipid conjugated to one or more N-acetylgalactosamine moieties. GlaNAc lipids optionally include additional moieties, such as, but not limited to, ethylene oxide groups, polyethylene oxide groups, alkylene groups, amide groups, ester groups, and the like. A “steroid” is a compound comprising the following carbon skeleton: . Non-limiting examples of steroids include cholesterol, and the like. A “cationic lipid” refers to a lipid capable of being positively charged. Exemplary cationic lipids include one or more amine group(s) which bear the positive charge. Preferred cationic lipids are ionizable such that they can exist in a positively charged or neutral form depending on pH. The ionization of the cationic lipid affects the surface charge of the lipid nanoparticle under different pH conditions. This charge state can influence plasma protein absorption, blood clearance and tissue distribution (Semple, S.C., et al., Adv. Drug Deliv Rev 32:3-17 (1998)) as well as the ability to form endosomolytic non-bilayer structures (Hafez, I.M., et al., Gene Ther 8:1188-1196 (2001)) critical to the intracellular delivery of nucleic acids. The term “lipid nanoparticle” or “LNP” refers to particles having at least one dimension on the order of nanometers (e.g., 1-1,000 nm), which may include one or more of the GalNAc lipids of the present disclosure (e.g., compounds of Formula (I)) or other specified GalNAc lipids). In some embodiments, LNPs are included in a 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 (e.g., cell, tissue, organ, tumor, and the like). In some embodiments, the LNPs of the disclosure comprise a nucleic acid. Such LNPs typically comprise a compound of Formula (I) and one or more components selected from cationic lipids, neutral lipids, charged lipids, steroids, and polymer conjugated lipids. In some embodiments, the active agent or therapeutic agent, such as a nucleic acid, may be encapsulated in the lipid portion of the lipid nanoparticle or an aqueous space enveloped by some or all the lipid portion of the lipid nanoparticle, thereby protecting it from enzymatic degradation or other undesirable effects induced by the mechanisms of the host organism or cells, e.g., an adverse immune response. In various embodiments, the LNPs have a mean diameter of from about 30 nm to about 150 nm, from about 40 nm to about 150 nm, from about 50 nm to about 150 nm, from about 60 nm to about 130 nm, from about 70 nm to about 110 nm, from about 70 nm to about 100 nm, from about 80 nm to about 100 nm, from about 90 nm to about 100 nm, from about 70 to about 90 nm, from about 80 nm to about 90 nm, from about 70 nm to about 80 nm, or about 30 nm, 35 nm, 40 nm, 45 nm, 50 nm, 55 nm, 60 nm, 65 nm, 70 nm, 75 nm, 80 nm, 85 nm, 90 nm, 95 nm, 100 nm, 105 nm, 110 nm, 115 nm, 120 nm, 125 nm, 130 nm, 135 nm, 140 nm, 145 nm, or 150 nm, and are substantially non-toxic. In certain embodiments, nucleic acids, when present in the LNPs, are resistant in aqueous solution to degradation with a nuclease. Lipid nanoparticles comprising nucleic acids and their method of preparation are disclosed in, e.g., U.S. Patent Publication Nos.2004 / 0142025, 2007 / 0042031 and PCT Pub. Nos. WO 2013 / 016058 and WO 2013 / 086373, the full disclosures of which are herein incorporated by reference in their entirety for all purposes. “GalNAc” refers to N-acetylgalactoseamine, i.e., 2-(Acetylamino)-2-deoxy-D-galactose. 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. A “GalNAc pegylated lipid” is a pegylated lipid that includes one or more GalNAc moieties. Pegylated lipids are known in the art and include 1-(monomethoxy-polyethyleneglycol)-2,3-dimyristoylglycerol (PEG-DMG) and the like. The term “neutral lipid” refers to any of several lipid species that exist either in an uncharged or neutral zwitterionic form at a selected pH. At physiological pH, such lipids include, but are not limited to, phosphotidylcholines such as 1,2-Distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-Dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-Dimyristoyl-sn-glycero-3- phosphocholine (DMPC), 1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2- dioleoyl-sn-glycero-3-phosphocholine (DOPC), phophatidylethanolamines such as 1,2-Dioleoyl- sn-glycero-3-phosphoethanolamine (DOPE), sphingomyelins (SM), ceramides, steroids such as sterols and their derivatives. Neutral lipids may be synthetic or naturally derived. The term “charged lipid” refers to any of a number of lipid species that exist in either a positively charged or negatively charged form independent of the pH within a useful physiological range, e.g., pH ~3 to pH ~9. Charged lipids may be synthetic or naturally derived. Examples of charged lipids include phosphatidylserines, phosphatidic acids, phosphatidylglycerols, phosphatidylinositols, sterol hemisuccinates, dialkyl trimethylammonium-propanes, (e.g., DOTAP, DOTMA), dialkyl dimethylaminopropanes, ethyl phosphocholines, dimethylaminoethane carbamoyl sterols (e.g., DC-Chol). As used herein, the term “aqueous solution” refers to a composition comprising water. “Alkyl” refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms that is saturated (i.e., contains no double and / or triple bonds), having from one to twenty-four carbon atoms (C1-C24 alkyl), one to sixteen carbon atoms (C1-C16 alkyl),one to twelve carbon atoms (C1-C12 alkyl), six to twenty-four carbon atoms (C6-C24 alkyl), one to eight carbon atoms (C1-C8 alkyl) or one to six carbon atoms (C1-C6 alkyl) and which is attached to the rest of the molecule by a single bond, e.g., methyl, ethyl, n-propyl, 1-methylethyl (iso propyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl), 3-methylhexyl, 2-methylhexyl, and the like. Unless stated otherwise specifically in the specification, an alkyl group is optionally substituted. “Alkenyl” refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms that contains at least one carbon-carbon double, having from one to twenty-four carbon atoms (C2-C24 alkenyl), one to twelve carbon atoms (C2-C12 alkenyl), six to twenty-four carbon atoms (C6-C24 alkenyl), two to sixteen carbon atoms (C2-C16 alkenyl), four to twelve carbon atoms (C4-C12 alkenyl), one to eight carbon atoms (C2-C8 alkenyl) or one to six carbon atoms (C2-C6 alkenyl) and which is attached to the rest of the molecule by a single bond, e.g., ethenyl, n-propenyl, 1-methylethenyl, n-butenyl, n-pentenyl, 1,1-dimethylethenyl, 3- methylhexenyl, 2-methylhexenyl, and the like. Unless stated otherwise specifically in the specification, an alkenyl group is optionally substituted. “Alkynyl” refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms that contains at least one carbon-carbon triple bond, having from one to twenty-four carbon atoms (C2-C24 alkynyl), one to twelve carbon atoms (C2-C12 alkynyl), one to eight carbon atoms (C2-C8 alkynyl) or one to six carbon atoms (C2-C6 alkynyl) and which is attached to the rest of the molecule by a single bond, e.g., ethynyl, n-propynyl, 1-methylethynyl, n-butynyl, n-pentynyl, 1,1-dimethylethynyl, 3-methylhexynyl, 2-methylhexynyl, and the like. Unless stated otherwise specifically in the specification, an alkynyl group is optionally substituted. “Alkylene” or “alkylene chain” refers to a straight or branched divalent hydrocarbon chain linking the rest of the molecule to a radical group, consisting solely of carbon and hydrogen and having, for example, from one to twenty-four carbon atoms (C1-C24 alkylene), one to fifteen carbon atoms (C1-C15 alkylene),one to twelve carbon atoms (C1-C12 alkylene), one to eight carbon atoms (C1-C8 alkylene), one to six carbon atoms (C1-C6 alkylene), two to four carbon atoms (C2-C4 alkylene), one to two carbon atoms (C1-C2 alkylene), e.g., methylene, ethylene, propylene, n-butylene, and the like. The alkylene chain is attached to the rest of the molecule through a single or double bond and to the radical group through a single or double bond. The points of attachment of the alkylene chain to the rest of the molecule and to the radical group can be through one carbon or any two carbons within the chain. Unless stated otherwise specifically in the specification, an alkylene chain may be optionally substituted. “Cycloalkyl” or “carbocyclic ring” refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon radical consisting solely of carbon and hydrogen atoms, which may include fused or bridged ring systems, having from three to fifteen carbon atoms, preferably having from three to ten carbon atoms, and which is saturated or unsaturated and attached to the rest of the molecule by a single bond. Monocyclic radicals include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic radicals include, for example, adamantyl, norbornyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. Unless otherwise stated specifically in the specification, a cycloalkyl group may be optionally substituted. “Cycloalkenyl” is a cycloalkyl group comprising at least one carbon-carbon double bond withing the cycloalkyl ring. Unless otherwise stated specifically in the specification, a cycloalkenyl group may be optionally substituted. “Cycloalkylene” is a divalent cycloalkyl group. Unless otherwise stated specifically in the specification, a cycloalkylene group may be optionally substituted. “Cycloalkenylene” is a divalent cycloalkenyl group. Unless otherwise stated specifically in the specification, a cycloalkenylene group may be optionally substituted. “Halo” refers to a halogen substituent (i.e., F, Cl, Br, or I). “Heterocyclyl” or “heterocyclic ring” refers to a stable 3- to 18-membered (e.g., 5, 6 or 7- membered) non-aromatic ring radical having one to twelve ring carbon atoms (e.g., two to twelve) and from one to six ring heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur. Unless stated otherwise specifically in the specification, the heterocyclyl radical may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include fused or bridged ring systems; and the nitrogen, carbon or sulfur atoms in the heterocyclyl radical may be optionally oxidized; the nitrogen atom may be optionally quaternized; and the heterocyclyl radical may be partially or fully saturated. Examples of such heterocyclyl radicals include, but are not limited to, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Unless stated otherwise specifically in the specification, a heterocyclyl group may be optionally substituted. "Heteroaryl" refers to a 5- to 14-membered ring system radical comprising hydrogen atoms, one to thirteen ring carbon atoms, one to six ring heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur, and at least one aromatic ring. Examples include, but are not limited to, pyrrolyl, imidazolyl, triazolyl, thiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, and the like. Unless stated otherwise specifically in the specification, a heteroaryl group may be optionally substituted. The term “substituted” used herein means any of the above groups (e.g., alkyl, alkenyl, alkynyl, alkylene, cycloalkyl, cycloalkenyl, cycloalkylene, cycloalkenylene, heterocyclyl and / or heteroaryl) wherein at least one hydrogen atom is replaced by a bond to a non-hydrogen atoms such as, but not limited to: a halogen atom, such as F, Cl, Br, and I, cyano, -OH, or - NH2.“Optional” or “optionally” (e.g., optionally substituted) means that the subsequently described event of circumstances may or may not occur, and that the description includes instances where said event or circumstance occurs and instances in which it does not. For example, “optionally substituted alkyl” means that the alkyl radical may or may not be substituted and that the description includes both substituted alkyl radicals and alkyl radicals having no substitution. In some embodiments, “optionally substituted” means a particular radical is substituted with one or more substituents selected from halo (e.g., F, Cl, Br, and I). This disclosure is also meant to encompass all pharmaceutically acceptable compounds of the present disclosure being isotopically labelled by having one or more atoms replaced by an atom having a different atomic mass or mass number. Examples of isotopes that can be incorporated into the disclosed compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, chlorine, and iodine, such as2H,3H,11C,13C,14C,13N,15N,15O,17O,18O,31P,32P,35S,18F,36Cl,123I, and125I, respectively. These radiolabeled compounds could be useful to help determine or measure the effectiveness of the compounds, by characterizing, for example, the site or mode of action, or binding affinity to pharmacologically important site of action. Certain isotopically labelled compounds of the present disclosure (e.g., compounds of Formula (I)), for example, those incorporating a radioactive isotope, are useful in drug and / or substrate tissue distribution studies. The radioactive isotopes tritium, i.e.,3H, and carbon-14, i.e.,14C, are particularly useful for this purpose in view of their ease of incorporation and ready means of detection. Substitution with heavier isotopes such as deuterium, i.e.,2H, may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances. Substitution with positron emitting isotopes, such as11C,18F,15O and13N, can be useful in Positron Emission Topography (PET) studies for examining substrate receptor occupancy. Isotopically labeled compounds of the present disclosure (e.g., compounds of Formula (I)) can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the Preparations and Examples as set out below using an appropriate isotopically labeled reagent in place of the non-labeled reagent previously employed. This disclosure is also meant to encompass the in vivo metabolic products of the disclosed compounds. Such products may result from, for example, the oxidation, reduction, hydrolysis, amidation, esterification, and the like of the administered compound, primarily due to enzymatic processes. Accordingly, the disclosure includes compounds produced by a process comprising administering a compound of this disclosure to a mammal for a period sufficient to yield a metabolic product thereof. Such products are typically identified by administering a radiolabeled compound of the present disclosure in a detectable dose to an animal, such as rat, mouse, guinea pig, monkey, or to human, allowing sufficient time for metabolism to occur, and isolating its conversion products from the urine, blood, or other biological samples. “Stable compound” and “stable structure” are meant to indicate a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent. “Mammal” includes humans and both domestic animals such as laboratory animals and household pets (e.g., cats, dogs, swine, cattle, sheep, goats, horses, rabbits), and non-domestic animals such as wildlife and the like. “Pharmaceutically acceptable carrier, diluent, or excipient” includes without limitation any adjuvant, carrier, excipient, glidant, sweetening agent, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier which has been approved by the United States Food and Drug Administration as being acceptable for use in humans or domestic animals. “Pharmaceutically acceptable salt” includes both acid and base addition salts. “Pharmaceutically acceptable acid addition salt” refers to those salts which retain the biological effectiveness and properties of the free bases, which are not biologically or otherwise undesirable, and which are formed with inorganic acids such as, but are not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, and organic acids such as, but not limited to, acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, camphoric acid, camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, 2- oxo-glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, undecylenic acid, and the like. “Pharmaceutically acceptable base addition salt” refers to those salts which retain the biological effectiveness and properties of the free acids, which are not biologically or otherwise undesirable. These salts are prepared from addition of an inorganic base or an organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, the sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts and the like. Preferred inorganic salts are the ammonium, sodium, potassium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, deanol, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, benethamine, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins and the like. Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline and caffeine. A “pharmaceutical composition” refers to a formulation of a compound of the present disclosure and a medium generally accepted in the art for the delivery of the biologically active compound to mammals, e.g., humans. Such a medium includes all pharmaceutically acceptable carriers, diluents, or excipients, therefore. An “effective amount” or “therapeutically effective amount” of an active agent or therapeutic agent such as a therapeutic nucleic acid is an amount sufficient to produce the desired effect, e.g., an increase or inhibition of expression of a target sequence in comparison to the normal expression level detected in the absence of the nucleic acid. An increase in expression of a target sequence is achieved when any measurable level is detected in the case of an expression product that is not present in the absence of the nucleic acid. In the case where the expression product is present at some level prior to contact with the nucleic acid, an in increase in expression is achieved when the fold increase in value obtained with a nucleic acid such as mRNA relative to control is about 1.05, 1.1, 1.2, 1.3, 1.4, 1.5, 1.75, 2, 2.5, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 75, 100, 250, 500, 750, 1000, 5000, 10000 or greater. Inhibition of expression of a target gene or target sequence is achieved when the value obtained with a nucleic acid such as antisense oligonucleotide relative to the control is about 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5%, or 0%. Suitable assays for measuring expression of a target gene or target sequence include, e.g., examination of protein or RNA levels using techniques known to those of skill in the art such as dot blots, northern blots, in situ hybridization, ELISA, immunoprecipitation, enzyme function, fluorescence or luminescence of suitable reporter proteins, as well as phenotypic assays known to those of skill in the art. “Treating” or “treatment” as used herein covers the treatment of the disease or condition of interest in a mammal, preferably a human, having the disease or condition of interest, and includes: (i) preventing the disease or condition from occurring in a mammal, in particular, when such mammal is predisposed to the condition but has not yet been diagnosed as having it; (ii) inhibiting the disease or condition, i.e., arresting its development; (iii) relieving the disease or condition, i.e., causing regression of the disease or condition; or (iv) relieving the symptoms resulting from the disease or condition, i.e., relieving pain without addressing the underlying disease or condition. As used herein, the terms “disease” and “condition” may be used interchangeably or may be different in that the particular malady or condition may not have a known causative agent (so that etiology has not yet been worked out) and it is therefore not yet recognized as a disease but only as an undesirable condition or syndrome, wherein a more or less specific set of symptoms have been identified by clinicians. The compounds of the present disclosure, or their pharmaceutically acceptable salts may contain one or more asymmetric centers and may thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that may be defined, in terms of absolute stereochemistry, as (R)- or (S)- or, as (D)- or (L)- for amino acids. The present disclosure is meant to include all such possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (-), (R)- and (S)-, or (D)- and (L)- isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, for example, chromatography and fractional crystallization. Conventional techniques for the preparation / isolation of individual enantiomers include chiral synthesis from a suitable optically pure precursor or resolution of the racemate (or the racemate of a salt or derivative) using, for example, chiral high-pressure liquid chromatography (HPLC). When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, and unless specified otherwise, it is intended that the compounds include both E and Z geometric isomers. Likewise, all tautomeric forms are also intended to be included. A “stereoisomer” refers to a compound made up of the same atoms bonded by the same bonds but having different three-dimensional structures, which are not interchangeable. The present disclosure contemplates various stereoisomers and mixtures thereof and includes “enantiomers”, which refers to two stereoisomers whose molecules are non-superimposable mirror images of one another. A “tautomer” refers to a proton shift from one atom of a molecule to another atom of the same molecule. The present disclosure includes tautomers of any said compounds. Compounds In an aspect, the disclosure provides novel GalNAc lipid compounds which can combine with other components such as cationic lipids, neutral lipids, charged lipids, steroids, and / or polymer conjugated lipids to form LNPs, which may optionally encapsulate therapeutic agents, such as oligonucleotides. Without wishing to be bound by theory, it is thought that these LNPs shield oligonucleotides from degradation in the serum and provide for effective delivery of oligonucleotides to cells in vitro and in vivo. In some embodiments is provided a compound having the structure of Formula (I): or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof (i.e., “compounds of the present disclosure”), wherein: R1, R2, and R3are each, independently L1, L2, L3, and L4are, at each occurrence, independently −NH(C=O)− or –(C=O)NH−; a is an integer from 4 to 12; b and c are each independently an integer from 1 to 12; d is an integer from 1 to 10; z has a mean value ranging from 1 to 60; and R4and R5are each independently a straight or branched alkyl, alkenyl or alkynyl chain containing from 10 to 30 carbon atoms, wherein the alkyl, alkenyl or alkynyl chain is optionally interrupted by one or more ester bonds. Compounds of Formula (I), and subembodiments thereof, are also referred to herein as “compounds of the present disclosure” and / or “GalNAc-pegylated lipids.” In certain embodiments, L1is −NH(C=O)−, wherein the nitrogen atom of L1is bound to the carbon bearing the R1, R2and R3groups. In other embodiments, L2is –(C=O)NH−, wherein the nitrogen atom of L2is bound to the carbon atom of the adjacent ethylene oxide group. In different embodiments, L3, at each occurrence, is –(C=O)NH−, wherein the carbon atom of L3is bound to the carbon atom of the group. In still other embodiments, L4, at each occurrence, is –NH(C=O)−, wherein the nitrogen atom of L4is bound to the carbon atom of In more specific embodiments, the compound has the structure of Formula (IA): or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof; wherein: R1, R2, and R3are each, independently a is an integer from 6 to 10; b and c are each independently an integer from 1 to 6; d is an integer from 1 to 5; z has a mean value ranging from 1 to 60; and R4and R5are each independently a straight or branched alkyl, alkenyl or alkynyl chain containing from 10 to 30 carbon atoms, wherein the alkyl, alkenyl or alkynyl chain is optionally interrupted by one or more ester bonds. In various embodiments, a is 8. In other embodiments, b and c are, at each occurrence, independently 3 or 4. In more embodiments, d is 1. In some other different embodiments, R4and R5are each independently straight alkyl chains containing from 12 to 18 carbon atoms. One other embodiment provides a compound having one of the following structures of Table 1: Table 1

[0003] or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein: z has a mean value ranging from 1 to 60. In some of the foregoing embodiments, z has a mean value ranging from 15 to 60, 30 to 60 or 40 to 50. In some embodiments, the mean value of z ranges from 40 to 55. In some embodiments, the mean value of z ranges from 40 to 50, or 42 to 48. In some embodiments, the mean value of z ranges from 30 to 55, 30 to 50, 30 to 45, 30 to 40, or 30 to 35. In some embodiments, the mean value of z ranges from 35 to 55, 40 to 55, 42 to 55, 45 to 55, or 48 to 55. In some of the foregoing embodiments, z spans a range that is selected such that the PEG portion of (I) or (IA) has an average molecular weight of about 400 to about 6000 g / mol. For example, in certain embodiments z has a mean value of about 45. In another embodiment, z has a mean value of about 48. One embodiment provides a compound having the following structure: or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof. In some embodiments, compounds of the present disclosure are optionally substituted. In certain embodiments, compounds of the present disclosure are optionally substituted with one or more substituents selected from the group consisting of halo, cyano, −OH, or −NH2. In certain embodiments, compounds of the present disclosure are substituted with one or more fluoro substituents. It is understood that any embodiment of the compounds of the present disclosure, as set forth above, and any specific substituent (e.g., an alkyl group) may be optionally substituted with one or more substituents (e.g., halo, −OH, −NH2). It is understood that such substitutions are permissible only if the substitution results in stable compounds. The compounds of the present disclosure may be used as components of LNPs, which in turn may be used for delivery of therapeutic agents, such as nucleic acids. The compounds of the present disclosure are present in the LNPs in an amount which is effective to form an LNP and deliver a therapeutic agent, e.g., for treating a particular disease or condition of interest. Appropriate concentration and dosages can be readily determined by one skilled in the art. One embodiment provides an LNP comprising a compound having the structure of Formula (I): or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof (i.e., “compounds of the present disclosure”); wherein: R1, R2, and R3are each, independently L1, L2, L3, and L4are, at each occurrence, independently −NH(C=O)− or –(C=O)NH−; a is an integer from 4 to 12; b and c are each independently an integer from 1 to 12; d is an integer from 1 to 10; z has a mean value ranging from 1 to 60; and R4and R5are each independently a straight or branched alkyl, alkenyl or alkynyl chain containing from 10 to 30 carbon atoms, wherein the alkyl, alkenyl or alkynyl chain is optionally interrupted by one or more ester bonds. In certain embodiments of the LNPs, L1is −NH(C=O)−, wherein the nitrogen atom of L1is bound to the carbon bearing the R1, R2and R3groups. In other LNP embodiments, L2is – (C=O)NH−, wherein the nitrogen atom of L2is bound to the carbon atom of the adjacent ethylene oxide group. In different embodiments of LNPs, L3, at each occurrence, is –(C=O)NH−, wherein the carbon atom of L3is bound to the carbon atom of the group. In still other embodiments of LNPs, L4, at each occurrence, is –NH(C=O)−, wherein the nitrogen atom of L4is bound to the carbon atom of the group. In more specific embodiments, the LNP comprises a compound having the structure of Formula (IA): or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof, wherein: R1, R2, and R3are each, independently a is an integer from 6 to 10; b and c are each independently an integer from 1 to 6; d is an integer from 1 to 5; z has a mean value ranging from 1 to 60; and R4and R5are each independently a straight or branched alkyl, alkenyl or alkynyl chain containing from 10 to 30 carbon atoms, wherein the alkyl, alkenyl or alkynyl chain is optionally interrupted by one or more ester bonds. In various embodiments of the LNP, a is 8. In other LNP embodiments, b and c are, at each occurrence, independently 3 or 4. In still more LNP embodiments, d is 1. In some other different embodiments of LNPs, R4and R5are each independently straight alkyl chains containing from 12 to 18 carbon atoms. One other embodiment provides an LNP comprising a compound having one of the following structures in Table 2: Table 2

[0004]

[0005] or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein z has a mean value ranging from 1 to 60. In some of the foregoing embodiments, z has a mean value ranging from 15 to 60, 30 to 60 or 40 to 50. In some embodiments, the mean value of z ranges from 40 to 55. In some embodiments, the mean value of z ranges from 40 to 50, or 42 to 48. In some embodiments, the mean value of z ranges from 30 to 55, 30 to 50, 30 to 45, 30 to 40, or 30 to 35. In some embodiments, the mean value of z ranges from 35 to 55, 40 to 55, 42 to 55, 45 to 55, or 48 to 55. In some of the foregoing embodiments, z spans a range that is selected such that the PEG portion of (I) or (IA) has an average molecular weight of about 400 to about 6000 g / mol. For example, in certain embodiments z has a mean value of about 45. In another embodiment, z has a mean value of 48. One embodiment provides a lipid nanoparticle that comprises a compound having the following structure:

[0006] or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof. In some embodiments the lipid nanoparticle further comprises a therapeutic agent selected from a nucleic acid. In certain embodiments, the nucleic acid is selected from antisense and messenger RNA. In other embodiments, the therapeutic agent comprises a Cas9 mRNA or a ribonucleoprotein. For example, the mRNA in some embodiments encodes and antigen (e.g., viral antigen) and the LNPs can be used for vaccinating against pathogens, such as viruses. Another embodiment provides a composition comprising a compound having the structure of Formula (I): or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof, and optionally lipid excipients and / or a therapeutic agent, wherein: R1, R2, and R3are each, independently: L1, L2, L3, and L4are, at each occurrence, independently −NH(C=O)− or –(C=O)NH−; a is an integer from 4 to 12; b and c are each independently an integer from 1 to 12; d is an integer from 1 to 10; z has a mean value ranging from 1 to 60; and R4and R5are each independently a straight or branched alkyl, alkenyl or alkynyl chain containing from 10 to 30 carbon atoms, wherein the alkyl, alkenyl or alkynyl chain is optionally interrupted by one or more ester bonds. In certain embodiments of the compositions, L1is −NH(C=O)−, wherein the nitrogen atom of L1is bound to the carbon bearing the R1, R2and R3groups. In other composition embodiments, L2is –(C=O)NH−, wherein the nitrogen atom of L2is bound to the carbon atom of the adjacent ethylene oxide group. In different embodiments of compositions, L3, at each occurrence, is –(C=O)NH−, wherein the carbon atom of L3is bound to the carbon atom of the group. In still other embodiments of compositions, L4, at each occurrence, is –NH(C=O)−, wherein the nitrogen atom of L4is bound to the carbon atom of the group. In more specific embodiments, the compositions comprise a compound having the structure of Formula (IA): or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof; wherein: R1, R2, and R3are each, independently: a is an integer from 6 to 10; b and c are each independently an integer from 1 to 6; d is an integer from 1 to 5; z has a mean value ranging from 1 to 60; and R4and R5are each independently a straight or branched alkyl, alkenyl or alkynyl chain containing from 10 to 30 carbon atoms, wherein the alkyl, alkenyl or alkynyl chain is optionally interrupted by one or more ester bonds. In various embodiments of the compositions, a is 8. In other composition embodiments, b and c are, at each occurrence, independently 3 or 4. In still more composition embodiments, d is 1. In some other different embodiments of compositions, R4and R5are each independently straight alkyl chains containing from 12 to 18 carbon atoms. One other embodiment provides a composition comprising a compound having one of the following structures in Table 3: Table 3

[0007] or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein z has a mean value ranging from 1 to 60, and optionally lipid excipients and / or a therapeutic agent. In some of the foregoing embodiments, z has a mean value ranging from 15 to 60, 30 to 60 or 40 to 50. In some embodiments, the mean value of z ranges from 40 to 55. In some embodiments, the mean value of z ranges from 40 to 50, or 42 to 48. In some embodiments, the mean value of z ranges from 30 to 55, 30 to 50, 30 to 45, 30 to 40, or 30 to 35. In some embodiments, the mean value of z ranges from 35 to 55, 40 to 55, 42 to 55, 45 to 55, or 48 to 55. In some of the foregoing embodiments, z spans a range that is selected such that the PEG portion of (I) or (IA) has an average molecular weight of about 400 to about 6000 g / mol. For example, in certain embodiments z has a mean value of about 45. In another embodiment, z has a mean value of 48. Another specific embodiment provides a composition that comprises a compound having the following structure:

[0008] or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof, optionally lipid excipients and / or a therapeutic agent. In some embodiments, the composition or LNP comprises a plurality of GalNAc- pegylated lipids of Formula (I). In some embodiments, the plurality of pegylated lipids has an average value of z ranging from 30 to 60. In some embodiments, the plurality of GalNAc- pegylated lipids has an average value of z ranging from 40 to 50, or 42 to 48. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z ranging from 30 to 55, 30 to 50, 30 to 45, 30 to 40, or 30 to 35. In some embodiments, the plurality of lipids has an average value of z ranging from 35 to 55, 40 to 55, 42 to 55, 45 to 55, or 48 to 55. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 30. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 31. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 32. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 33. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 34. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 35. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 36. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 37. In some embodiments, the plurality of GalNAc- pegylated lipids has an average value of z of about 38. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 39. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 40. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 41. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 42. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 43. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 44. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 45. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 46. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 47. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 48. In some embodiments, the plurality of GalNAc- pegylated lipids has an average value of z of about 49. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 50. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 51. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 52. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 53. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 54. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 55. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 56. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 57. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 58. In some embodiments, the plurality of GalNAc-pegylated lipids has an average value of z of about 59. In some embodiments, the plurality of GalNAc- pegylated lipids has an average value of z of about 60. In some of the forgoing embodiments, the composition comprises a therapeutic agent selected from a nucleic acid. In certain embodiments, the nucleic acid is selected from antisense and messenger RNA. In other embodiments, the therapeutic agent comprises a Cas9 mRNA or a ribonucleoprotein. In some embodiments, the compositions or LNPs comprising compounds of the present disclosure further comprises a cationic lipid. Exemplary cationic lipids for use in the present disclosure include those disclosed in PCT Publication Nos. WO 2015 / 199952; WO 2017 / 004143; WO 2017 / 075531; WO 2017 / 117528; WO 2018 / 191657; WO 2018 / 107026; WO 2018 / 200943; WO 2018 / 078053; WO 2019 / 036000; WO 2019 / 036028; WO 2019 / 036030; WO 2019 / 036008; WO 2020 / 061426; WO 2020 / 081938; WO 2020 / 146805; WO 2021 / 030701; WO 2022 / 016070; WO 2023 / 114944; WO 2023 / 114937; WO 2023 / 114943; WO 2023 / 250427; and WO 2024 / 054843, which are incorporate herein by reference in its entirety. For example, some embodiments of the compositions or LNPs further comprise at least one cationic lipid having a structure of Formula (IIA) or (IIB):

[0009] or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof, wherein: L21and L22are each independently −O(C=O)−, −(C=O)O−, −C(=O)−, −O−, −S(O)x−, −S−S−, −C(=O)S−, −SC(=O)−, −NRaC(=O)−, −C(=O)NRa−, −NRaC(=O)NRa−, −OC(=O)NRa− or −NRaC(=O)O−; G23is C1-C6 alkylene; Rais H or C1-C12alkyl; R21aand R21bare, at each occurrence, independently either: (a) H or C1-C12 alkyl; or (b) R21ais H or C1-C12 alkyl, and R21btogether with the carbon atom to which it is bound is taken together with an adjacent R21band the carbon atom to which it is bound to form a carbon- carbon double bond; R22aand R22bare, at each occurrence, independently either: (a) H or C1-C12 alkyl; or (b) R22ais H or C1-C12alkyl, and R22btogether with the carbon atom to which it is bound is taken together with an adjacent R22band the carbon atom to which it is bound to form a carbon- carbon double bond; R23aand R23bare, at each occurrence, independently either (a): H or C1-C12 alkyl; or (b) R23ais H or C1-C12alkyl, and R23btogether with the carbon atom to which it is bound is taken together with an adjacent R23band the carbon atom to which it is bound to form a carbon- carbon double bond; R24aand R24bare, at each occurrence, independently either: (a) H or C1-C12 alkyl; or (b) R24ais H or C1-C12 alkyl, and R24btogether with the carbon atom to which it is bound is taken together with an adjacent R24band the carbon atom to which it is bound to form a carbon- carbon double bond; R25and R26are each independently H or methyl; R27is C6-C16alkyl; R28and R29are each independently C1-C12 alkyl; or R28and R29, together with the nitrogen atom to which they are attached, form a 5, 6 or 7-membered heterocyclic ring; e′, f′, g′, and h′ are each independently an integer from 1 to 24; and x is 0, 1 or 2. In some embodiments, the cationic lipid has structure (IIA). In other embodiments, the cationic lipid has structure (IIB). In any of the foregoing embodiments, one of L21or L22is -O(C=O)-. For example, in some embodiments each of L21and L22are -O(C=O)-. In some different embodiments of any of the foregoing, one of L21or L22is -(C=O)O-. For example, in some embodiments each of L21and L22is -(C=O)O-. In different embodiments, one of L21or L22is a direct bond. As used herein, a “direct bond” means the group (e.g., L21or L22) is absent. For example, in some embodiments each of L21and L22is a direct bond. In other different embodiments of the foregoing, for at least one occurrence of R21aand R21b, R21ais H or C1-C12 alkyl, and R21btogether with the carbon atom to which it is bound is taken together with an adjacent R21band the carbon atom to which it is bound to form a carbon- carbon double bond. In still other different embodiments, for at least one occurrence of R24aand R24b, R24ais H or C1-C12 alkyl, and R24btogether with the carbon atom to which it is bound is taken together with an adjacent R24band the carbon atom to which it is bound to form a carbon-carbon double bond. In more embodiments, for at least one occurrence of R22aand R22b, R22ais H or C1-C12 alkyl, and R22btogether with the carbon atom to which it is bound is taken together with an adjacent R22band the carbon atom to which it is bound to form a carbon-carbon double bond. In other different embodiments of any of the foregoing, for at least one occurrence of R23aand R23b, R23ais H or C1-C12 alkyl, and R23btogether with the carbon atom to which it is bound is taken together with an adjacent R23band the carbon atom to which it is bound to form a carbon- carbon double bond. It is understood that “carbon-carbon” double bond refers to one of the following structures: wherein Rcand Rdare, at each occurrence, independently H or a substituent. For example, in some embodiments Rcand Rdare, at each occurrence, independently H, C1-C12 alkyl or cycloalkyl, for example H or C1-C12 alkyl. In various other embodiments, the compound has one of the following structures (IIC) or (IID): wherein e, f, g and h are each independently an integer from 1 to 12. In some embodiments, the cationic lipid has structure (IIC). In other embodiments, the cationic lipid has structure (IID). In various embodiments of the cationic lipid of structures (IIC) or (IID), e, f, g and h are each independently an integer from 4 to 10. In certain embodiments of the foregoing, a, b, c and d are each independently an integer from 2 to 12 or an integer from 4 to 12. In other embodiments, a, b, c and d are each independently an integer from 8 to 12 or 5 to 9. In some certain embodiments, a is 0. In some embodiments, a is 1. In other embodiments, a is 2. In more embodiments, a is 3. In yet other embodiments, a is 4. In some embodiments, a is 5. In other embodiments, a is 6. In more embodiments, a is 7. In yet other embodiments, a is 8. In some embodiments, a is 9. In other embodiments, a is 10. In more embodiments, a is 11. In yet other embodiments, a is 12. In some embodiments, a is 13. In other embodiments, a is 14. In more embodiments, a is 15. In yet other embodiments, a is 16. In some embodiments, b is 1. In other embodiments, b is 2. In more embodiments, b is 3. In yet other embodiments, b is 4. In some embodiments, b is 5. In other embodiments, b is 6. In more embodiments, b is 7. In yet other embodiments, b is 8. In some embodiments, b is 9. In other embodiments, b is 10. In more embodiments, b is 11. In yet other embodiments, b is 12. In some embodiments, b is 13. In other embodiments, b is 14. In more embodiments, b is 15. In yet other embodiments, b is 16. In some embodiments, c is 1. In other embodiments, c is 2. In more embodiments, c is 3. In yet other embodiments, c is 4. In some embodiments, c is 5. In other embodiments, c is 6. In more embodiments, c is 7. In yet other embodiments, c is 8. In some embodiments, c is 9. In other embodiments, c is 10. In more embodiments, c is 11. In yet other embodiments, c is 12. In some embodiments, c is 13. In other embodiments, c is 14. In more embodiments, c is 15. In yet other embodiments, c is 16. In some certain embodiments, d is 0. In some embodiments, d is 1. In other embodiments, d is 2. In more embodiments, d is 3. In yet other embodiments, d is 4. In some embodiments, d is 5. In other embodiments, d is 6. In more embodiments, d is 7. In yet other embodiments, d is 8. In some embodiments, d is 9. In other embodiments, d is 10. In more embodiments, d is 11. In yet other embodiments, d is 12. In some embodiments, d is 13. In other embodiments, d is 14. In more embodiments, d is 15. In yet other embodiments, d is 16. In some embodiments, e is 1. In other embodiments, e is 2. In more embodiments, e is 3. In yet other embodiments, e is 4. In some embodiments, e is 5. In other embodiments, e is 6. In more embodiments, e is 7. In yet other embodiments, e is 8. In some embodiments, e is 9. In other embodiments, e is 10. In more embodiments, e is 11. In yet other embodiments, e is 12. In some embodiments, f is 1. In other embodiments, f is 2. In more embodiments, f is 3. In yet other embodiments, f is 4. In some embodiments, f is 5. In other embodiments, f is 6. In more embodiments, f is 7. In yet other embodiments, f is 8. In some embodiments, f is 9. In other embodiments, f is 10. In more embodiments, f is 11. In yet other embodiments, f is 12. In some embodiments, g is 1. In other embodiments, g is 2. In more embodiments, g is 3. In yet other embodiments, g is 4. In some embodiments, g is 5. In other embodiments, g is 6. In more embodiments, g is 7. In yet other embodiments, g is 8. In some embodiments, g is 9. In other embodiments, g is 10. In more embodiments, g is 11. In yet other embodiments, g is 12. In some embodiments, h is 1. In other embodiments, e is 2. In more embodiments, h is 3. In yet other embodiments, h is 4. In some embodiments, e is 5. In other embodiments, h is 6. In more embodiments, h is 7. In yet other embodiments, h is 8. In some embodiments, h is 9. In other embodiments, h is 10. In more embodiments, h is 11. In yet other embodiments, h is 12. In some other various embodiments, a and d are the same. In some other embodiments, b and c are the same. In some other specific embodiments and a and d are the same and b and c are the same. The sum of a and b and the sum of c and d are factors which may be varied to obtain a lipid having the desired properties. In one embodiment, a and b are chosen such that their sum is an integer ranging from 14 to 24. In other embodiments, c and d are chosen such that their sum is an integer ranging from 14 to 24. In further embodiment, the sum of a and b and the sum of c and d are the same. For example, in some embodiments the sum of a and b and the sum of c and d are both the same integer which may range from 14 to 24. In still more embodiments, a. b, c and d are selected such that the sum of a and b and the sum of c and d is 12 or greater. The substituents at R21a, R22a, R23aand R24aare not particularly limited. In some embodiments, at least one of R21a, R22a, R23aand R24ais H. In certain embodiments R21a, R22a, R23aand R24aare H at each occurrence. In certain other embodiments at least one of R21a, R22a, R23aand R24ais C1-C12 alkyl. In certain other embodiments at least one of R21a, R22a, R23aand R24ais C1-C8 alkyl. In certain other embodiments at least one of R21a, R22a, R23aand R24ais C1-C6 alkyl. In some of the foregoing embodiments, the C1-C8 alkyl is methyl, ethyl, n-propyl, iso- propyl, n-butyl, iso-butyl, tert-butyl, n-hexyl or n-octyl. In certain embodiments of the foregoing, R21a, R21b, R24aand R24bare C1-C12 alkyl at each occurrence. In further embodiments of the foregoing, at least one of R21b, R22b, R23band R24bis H or R21b, R22b, R23band R24bare H at each occurrence. In certain embodiments of the foregoing, R21btogether with the carbon atom to which it is bound is taken together with an adjacent R21band the carbon atom to which it is bound to form a carbon-carbon double bond. In other embodiments of the foregoing R24btogether with the carbon atom to which it is bound is taken together with an adjacent R24band the carbon atom to which it is bound to form a carbon-carbon double bond. The substituents at R25and R26are not particularly limited in the foregoing embodiments. In certain embodiments one of R25or R26is methyl. In other embodiments each of R25or R26is methyl. The substituents at R27are not particularly limited in the foregoing embodiments. In certain embodiments R27is C6-C16 alkyl. In some other embodiments, R27is C6-C9 alkyl. In some of these embodiments, R27is substituted with -(C=O)ORb, –O(C=O)Rb, -C(=O)Rb, -ORb, -S(O)xRb, -S-SRb, -C(=O)SRb, -SC(=O)Rb, -NRaRb, -NRaC(=O)Rb, -C(=O)NRaRb, -NRaC(=O)NRaRb, -OC(=O)NRaRb, -NRaC(=O)ORb, -NRaS(O)xNRaRb, -NRaS(O)xRbor -S(O)xNRaRb, wherein: Rais H or C1-C12 alkyl; Rbis C1-C15 alkyl; and x is 0, 1 or 2. For example, in some embodiments R27is substituted with -(C=O)ORbor –O(C=O)Rb. In various of the foregoing embodiments, Rbis branched C1-C15 alkyl. For example, in some embodiments Rbhas one of the following structures: . In certain other of the foregoing embodiments, one of R28or R29is methyl. In other embodiments, both R28and R29are methyl. In some different embodiments, R28and R29, together with the nitrogen atom to which they are attached, form a 5, 6 or 7-membered heterocyclic ring. In some embodiments of the foregoing, R28and R29, together with the nitrogen atom to which they are attached, form a 5- membered heterocyclic ring, for example a pyrrolidinyl ring. In some different embodiments of the foregoing, R28and R29, together with the nitrogen atom to which they are attached, form a 6- membered heterocyclic ring, for example a piperazinyl ring. In still other embodiments of the foregoing compounds, G23is C2-C4 alkylene, for example C3 alkylene. In some embodiments, the compositions or lipid nanoparticles comprise a GalNAc- pegylated lipid of Formula (I) and further comprise at least one cationic lipid selected from Table 4. Table 4: Exemplary Cationic Lipids

[0010] In one specific example, the cationic lipid is bis(2-butyloctyl) 10-(N-(3-(pyrrolidin-1- yl)propyl)nonanamido)nonadecanedioate, the structure of which is provided below: . In some other embodiments, the compositions or LNPs comprise a GalNAc lipid compound of structure (I) and further comprise at least one cationic lipid having a structure of Formula (III): III or a pharmaceutically acceptable salt, tautomer, prodrug or stereoisomer thereof, wherein: one of L31or L32is –O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O)x-, -S-S-, -C(=O)S-, SC(=O)-, -NRaC(=O)-, -C(=O)NRa-, NRaC(=O)NRa-, -OC(=O)NRa- or -NRaC(=O)O-, and the other of L1or L2is –O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O)x-, -S-S-, -C(=O)S-, SC(=O)-, -NRaC(=O)-, -C(=O)NRa-, ,NRaC(=O)NRa-, -OC(=O)NRa- or -NRaC(=O)O- or a direct bond; G1and G2are each independently unsubstituted C1-C12 alkylene or C1-C12 alkenylene; G3is C1-C24 alkylene, C1-C24 alkenylene, C3-C8 cycloalkylene, C3-C8 cycloalkenylene; Rais H or C1-C12 alkyl; R31and R32are each independently C6-C24 alkyl or C6-C24 alkenyl; R33is H, OR35, CN, -C(=O)OR34, -OC(=O)R34or –NR35C(=O)R34; R34is C1-C12 alkyl; R35is H or C1-C6 alkyl; and x is 0, 1 or 2. In some of the foregoing embodiments, the compound has one of the following structures (IIIA) or (IIIB): wherein: A is a 3 to 8-membered cycloalkyl or cycloalkylene ring; R36is, at each occurrence, independently H, OH or C1-C24 alkyl; n is an integer ranging from 1 to 15. In some of the foregoing embodiments, the compound has structure (IIIA), and in other embodiments, the compound has structure (IIIB). In other embodiments of the foregoing, the compound has one of the following structures (IIIC) or (IIID): wherein y and z are each independently integers ranging from 1 to 12. In any of the foregoing embodiments, one of L31or L32is -O(C=O)-. For example, in some embodiments each of L31and L32are -O(C=O)-. In some different embodiments of any of the foregoing, L31and L32are each independently -(C=O)O- or -O(C=O)-. For example, in some embodiments each of L31and L32is -(C=O)O-. In some different embodiments of the foregoing, the compound has one of the following structures (IIIE) or (IIIF): In some of the foregoing embodiments, the compound has one of the following structures In some of the foregoing embodiments, n is an integer ranging from 2 to 12, for example from 2 to 8 or from 2 to 4. For example, in some embodiments, n is 3, 4, 5 or 6. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, n is 6. In some other of the foregoing embodiments, y and z are each independently an integer ranging from 2 to 10. For example, in some embodiments, y and z are each independently an integer ranging from 4 to 9 or from 4 to 6. In some of the foregoing embodiments, R36is H. In other of the foregoing embodiments, R36is C1-C24 alkyl. In other embodiments, R36is OH. In some embodiments, G33is unsubstituted. In other embodiments, G33is substituted. In various different embodiments, G33is linear C1-C24 alkylene or linear C1-C24 alkenylene. In some other foregoing embodiments, R31or R32, or both, is C6-C24 alkenyl. For example, in some embodiments, R31and R32each, independently have the following structure: , wherein: R37aand R37bare, at each occurrence, independently H or C1-C12 alkyl; and a is an integer from 2 to 12, wherein R37a, R37band a are each selected such that R31and R32each independently comprise from 6 to 20 carbon atoms. For example, in some embodiments a is an integer ranging from 5 to 9 or from 8 to 12. In some of the foregoing embodiments, at least one occurrence of R37ais H. For example, in some embodiments, R37ais H at each occurrence. In other different embodiments of the foregoing, at least one occurrence of R37bis C1-C8 alkyl. For example, in some embodiments, C1-C8 alkyl is methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, tert-butyl, n-hexyl or n- octyl. In different embodiments, R31or R32, or both, has one of the following structures: In some of the foregoing embodiments, R33is OH, CN, -C(=O)OR34, -OC(=O)R34or –NHC(=O)R34. In some embodiments, R34is methyl or ethyl. In various different embodiments, the cationic lipid has one of the structures set forth in Table 5 below. Table 5: Exemplary Cationic Lipids

[0011] In certain embodiments, the cationic lipid is ((3-hydroxypropyl)azanediyl)bis(nonane- 9,1-diyl) bis(2-butyloctanoate). That is, in some embodiments, the cationic lipid has the following structure: In still other embodiments, the LNPs and compositions comprise a GalNAc lipid compound of structure (I) and further comprise at least one cationic lipid having a structure of Formula (IV): or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof, wherein: R41is optionally substituted C1-C24 alkyl or optionally substituted C2-C24 alkenyl; R42and R43are each independently optionally substituted C1-C36 alkyl; R44and R45are each independently optionally substituted C1-C6 alkyl, or R44and R45join, along with the N to which they are attached, to form a heterocyclyl or heteroaryl; L41, L42, and L43are each independently optionally substituted C1-C18 alkylene; G41is a direct bond, -(CH2)nO(C=O)-, -(CH2)n(C=O)O-, or –(C=O)-; G42and G43are each independently -(C=O)O- or -O(C=O)-; and n is an integer greater than 0. In some embodiments, the compound has the following structure (IVA): In some embodiments, the compound has the following structure (IVB): In some embodiments, R41is optionally substituted C6-C18 alkyl or C14-C18 alkenyl. In certain embodiments, R41is C8 alkyl, C9 alkyl, C10 alkyl, C12 alkyl, C14 alkyl, or C16 alkyl. In some more specific embodiments, R41is C16 alkenyl. In certain more specific embodiments, R41is unbranched. In some embodiments, R41is branched. In certain embodiments, R41is unsubstituted. In some embodiments, G41is a direct bond, -(CH2)nO(C=O)-, or -(CH2)n(C=O)O-. In certain embodiments, G41is a direct bond. In some more specific embodiments, G41is - (CH2)n(C=O)O- and n is greater than 1. In some embodiments, n is 1-20. In some embodiments n is 1-10. In some embodiments n is 5-11. In some embodiments, n is 6-10. In certain more specific embodiments, n is 5, 6, 7, 8, 9, or 10. In some embodiments, n is 5. In some embodiments, n is 6. In some embodiments, n is 7. In certain embodiments, n is 8. In some embodiments, n is 9. In some embodiments, n is 10. In some embodiments, L41is C1-C6 alkylene. In certain embodiments, L41is C2 alkylene, C3 alkylene, or C4 alkylene. In some more specific embodiments, L41is unbranched. In certain more specific embodiments, L41is unsubstituted. In some embodiments, R42is C8-C24 alkyl. In some embodiments, R43is C8-C24 alkyl. In some more specific embodiments, R42and R43are both C8-C24 alkyl. In some embodiments, R42and R43are each independently C11 alkyl, C12 alkyl, C13 alkyl, C14 alkyl, C15 alkyl, C16 alkyl, C18 alkyl, or C20 alkyl. In certain embodiments, R42is branched. In more specific embodiments, R43is branched. In some more specific embodiments, R42and R43each independently have one of the following structures: wherein: R46and R47are each independently C2-C12 alkyl. In some embodiments, R42and R43each independently have one of the following structures: In some embodiments, L42and L43are each independently C4-C10 alkylene. In certain embodiments, L42and L43are both C5 alkylene. In some more specific embodiments, L42and L43are both C6 alkylene. In certain embodiments, L42and L43are both C8 alkylene. In some more specific embodiments, L42and L43are both C9 alkylene. In some embodiments, L42is unbranched. In some embodiments, L43is unbranched. In more specific embodiments, L42is unsubstituted. In some embodiments, L42is unsubstituted. In some embodiments, R44and R45are each independently C1-C6 alkyl. In more specific embodiments, R44and R45are both methyl. In certain embodiments, R44and R45are both ethyl. In certain embodiments, R44is methyl and R45is n-butyl. In some embodiments, R44and R45are both n-butyl. In different embodiments, R44is methyl and R45is n-hexyl. In some embodiments, R44and R45join, along with the N to which they are attached, to form a heterocyclyl. In certain embodiments, the heterocyclyl is a 5-membered heterocyclyl. In some embodiments, the heterocyclyl has the following structure: . In various different embodiments, the compound of structure IV has one of the structures set forth in Table 6 below. Table 6: Exemplary Cationic Lipids

[0012] In some embodiments, the cationic lipid is bis(2-butyloctyl) 10-(N-decyl-4- (dimethylamino)butanamido)nonadecanedioate. That is, in some embodiments, the cationic lipid has the following structure: Further exemplary cationic lipids and their synthesis can also be found in the following publications: US Patent Nos. US 9,738,593; US 10,221,127; US 10,166,298; US 11,357,856; US 11,712,481; and US 11,453,639; US Patent Publication Nos: US 2018 / 0185516 and US 2022 / 0106257; and PCT Publication Nos. WO 2017 / 117528; WO 2016 / 176330; WO 2018 / 191719; WO 2018 / 200943; WO 2019 / 036000; WO 2019 / 036028; WO 2019 / 036030; WO 2019 / 036008; WO 2019 / 089828; WO 2020 / 061426; WO 2020 / 081938; WO 2021 / 030701; WO 2023 / 114944; WO 2023 / 114939; WO 2023 / 114943, the disclosures of which are hereby incorporated by reference. In some embodiments, the composition or LNP further comprises a neutral lipid. In some embodiments, the composition or LNP further comprises a steroid. In some embodiments, the composition or LNP further comprises an additional polymer conjugated lipid (e.g., a pegylated lipid). In some embodiments, the composition or LNP comprising a compound of the present disclosure further comprises a neutral lipid, a steroid, and / or an additional polymer conjugated lipid (e.g., pegylated lipid), or a combination thereof. In some embodiments, the composition or LNP comprising a compound of the present disclosure further comprises an additional cationic lipid, a neutral lipid, a steroid, and / or an additional polymer conjugated lipid (e.g., pegylated lipid), or a combination thereof. One embodiment provides a composition or lipid nanoparticle comprising a compound of the present disclosure and at least one neutral lipid selected from the group consisting of 1,2- Distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-Dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-Dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1-Palmitoyl-2-oleoyl-sn- glycero-3-phosphocholine (POPC), l,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2- Dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), a sphingomyelin (SM), and combinations thereof. In some embodiments, the at least one neutral lipid comprises 1,2-Distearoyl-sn-glycero- 3-phosphocholine (DSPC). In some embodiments, the neutral lipid is present at a concentration ranging from about 5 to about 15 mol% of the lipid nanoparticle. As used herein, “mol percent,” “mole percent,” or “mol%” refers to a component’s molar percentage relative to the total number of mols of all components of a lipid nanoparticle excluding a therapeutic agent (i.e., total mols of compound of the present disclosure, cationic lipid(s), neutral lipid(s), steroid(s), and / or polymer conjugated lipid(s)). In some embodiments, the molar ratio of the cationic lipid to the neutral lipid ranges from about 2:1 to 8:1. In certain embodiments, the LNPs or compositions comprise a compound of the present disclosure and an additional polymer conjugated lipid, for example an additional pegylated lipid. For example, some embodiments include a pegylated diacylglycerol (PEG-DAG) such as 1-(monomethoxy-polyethyleneglycol)-2,3-dimyristoylglycerol (PEG-DMG), a pegylated phosphatidylethanoloamine (PEG-PE), a PEG succinate diacylglycerol (PEG-S-DAG) such as 4- O-(2’,3’-di(tetradecanoyloxy)propyl-1-O-(^-methoxy(polyethoxy)ethyl)butanedioate (PEG-S- DMG), a pegylated ceramide (PEG-cer), or a PEG dialkoxypropylcarbamate such as ^- methoxy(polyethoxy)ethyl-N-(2,3-di(tetradecanoxy)propyl)carbamate or 2,3- di(tetradecanoxy)propyl-N-(^-methoxy(polyethoxy)ethyl)carbamate. In some embodiments, the pegylated lipid is PEG-DMG. In some embodiments, the composition or LNP further comprises at least one additional pegylated lipid having a structure of Formula (V): or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein: R51and R52are each independently a straight or branched alkyl, alkenyl, or alkynyl containing from 10 to 30 carbon atoms, wherein each alkyl, alkenyl, or alkynyl is optionally substituted with at least one fluoro; and z′ is an integer ranging from 30 to 60. In some embodiments, R51and R52are each independently straight, alkyl chains containing from 12 to 16 carbon atoms, wherein each alkyl is optionally substituted with at least one fluoro. In certain embodiments, R31and R32are each independently straight alkyl chains containing from 12 to 16 carbon atoms. In some embodiments, R51and R52are each independently: . In some embodiments, wherein z′ is an integer ranging from 45 to 50. In some embodiments, wherein z is an integer ranging from 42 to 48. In some embodiments, the at least one additional pegylated lipid has the following structure: or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof. In some embodiments, the composition or LNP comprises a compound of the present disclosure and a plurality of pegylated lipids of Formula (V). In some embodiments, the plurality of pegylated lipids has an average value of z′ ranging from 40 to 55. In some embodiments, the plurality of pegylated lipids has an average value of z′ ranging from 40 to 50, or 42 to 48. In some embodiments, the plurality of pegylated lipids has an average value of z′ ranging from 30 to 55, 30 to 50, 30 to 45, 30 to 40, or 30 to 35. In some embodiments, the plurality of lipids has an average value of z′ ranging from 35 to 55, 40 to 55, 42 to 55, 45 to 55, or 48 to 55. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 30. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 31. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 32. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 33. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 34. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 35. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 36. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 37. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 38. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 39. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 40. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 41. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 42. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 43. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 44. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 45. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 46. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 47. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 48. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 49. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 50. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 51. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 52. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 53. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 54. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 55. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 56. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 57. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 58. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 59. In some embodiments, the plurality of pegylated lipids has an average value of z′ of about 60. Synthesis of pegylated lipids (e.g., compounds of Formula (V)) can be found in US Patent No.9,738,593, the disclosure of which is hereby incorporated by reference. In some embodiments, a compound of the present disclosure is present at a concentration ranging from about 0.05 to about 0.20 mol% of the LNP. In some embodiments, a compound of the present disclosure is present at a concentration ranging from about 0.05 to about 0.15 mol%, from about 0.06 to about 0.15 mol%, from about 0.06 to about 0.10 mol%, from about 0.10 to about 0.15 mol%, or from about 0.07 to about 0.13 mol% of the lipid nanoparticle. In some embodiments, a compound of the present disclosure is present at a concentration of about 0.05 mol% of the LNP. In some embodiments, a compound of the present disclosure is present at a concentration of about 0.06 mol% of the LNP. In some embodiments, a compound of the present disclosure is present at a concentration of about 0.07 mol% of the LNP. In some embodiments, a compound of the present disclosure is present at a concentration of about 0.075 mol% of the LNP. In some embodiments, a compound of the present disclosure is present at a concentration of about 0.08 mol% of the LNP. In some embodiments, a compound of the present disclosure is present at a concentration of about 0.09 mol% of the LNP. In some embodiments, a compound of the present disclosure is present at a concentration of about 0.10 mol% of the LNP. In some embodiments, a compound of the present disclosure is present at a concentration of about 0.11 mol% of the LNP. In some embodiments, a compound of the present disclosure is present at a concentration of about 0.12 mol% of the LNP. In some embodiments, a compound of the present disclosure is present at a concentration of about 0.13 mol% of the LNP. In some embodiments, a compound of the present disclosure is present at a concentration of about 0.14 mol% of the LNP. In some embodiments, a compound of the present disclosure is present at a concentration of about 0.15 mol% of the LNP. In some embodiments, a compound of the present disclosure is present at a concentration of about 0.16 mol% of the LNP. In some embodiments, a compound of the present disclosure is present at a concentration of about 0.17 mol% of the LNP. In some embodiments, a compound of the present disclosure is present at a concentration of about 0.18 mol% of the LNP. In some embodiments, a compound of the present disclosure is present at a concentration of about 0.19 mol% of the LNP. In some embodiments, a compound of the present disclosure is present at a concentration of about 0.20 mol% of the LNP. In some embodiments, a compound of the present disclosure is present at a concentration of about 0.21 mol% of the LNP. In some embodiments, a compound of the present disclosure is present at a concentration of about 0.22 mol% of the LNP. In some embodiments, a compound of the present disclosure is present at a concentration of about 0.23 mol% of the LNP. In some embodiments, a compound of the present disclosure is present at a concentration of about 0.24 mol% of the LNP. In some embodiments, a compound of the present disclosure is present at a concentration of about 0.25 mol% of the LNP. In some embodiments, the molar ratio of the cationic lipid to the compound of the present disclosure ranges from about 1000:1 to about 100:1 or from about 700:1 to about 200:1. In some embodiments, the concentration of the additional pegylated lipid, when present, ranges from about 1.0 to about 10.0 mol% of the lipid nanoparticle. In some embodiments, the concentration of the additional pegylated lipid (e.g., compounds of Formula (V)) ranges from about 2.0 to about 3.0 mol% of the LNP. In some embodiments, the concentration of the additional pegylated lipid (e.g., compounds of Formula (V)) is about 2.7 mol% of the LNP. In certain embodiments, the concentration of the additional pegylated lipid (e.g., compounds of Formula (V)) is about 2.4 or 2.7 mol % of the LNP. In some embodiments, the combined concentration of an additional pegylated lipid (e.g., compounds of Formula (V)), when present, and a compound of the present disclosure is about 1.0 to 5.0 mol% of the LNP. In some embodiments, the combined concentration of an additional pegylated lipid (e.g., compounds of Formula (V)), when present, and a compound of the present disclosure is about 2.0 to 3.0 mol% of the LNP. In some embodiments, the combined concentration of an additional pegylated lipid (e.g., compounds of Formula (V)), when present, and a compound of the present disclosure is about 2.2 to 2.8 mol% of the LNP. In some embodiments, the combined concentration of an additional pegylated lipid (e.g., compounds of Formula (V)), when present, and a compound of the present disclosure is about 2.3 to 2.7 mol% of the LNP. In some embodiments, the combined concentration of an additional pegylated lipid (e.g., compounds of Formula (V)), when present, and a compound of the present disclosure is about 2.4 to 2.6 mol% of the LNP. In some embodiments, the combined concentration of an additional pegylated lipid (e.g., compounds of Formula (V)), when present, and a compound of the present disclosure is about 2.4 mol% of the LNP. In some embodiments, the combined concentration of an additional pegylated lipid (e.g., compounds of Formula (V)), when present, and a compound of the present disclosure is about 2.45 mol% of the LNP. In some embodiments, the combined concentration of an additional pegylated lipid (e.g., compounds of Formula (V)), when present, and a compound of the present disclosure is about 2.5 mol% of the LNP. In some embodiments, the combined concentration of an additional pegylated lipid (e.g., compounds of Formula (V)), when present, and a compound of the present disclosure is about 2.55 mol% of the LNP. In some embodiments, the combined concentration of an additional pegylated lipid (e.g., compounds of Formula (V)), when present, and a compound of the present disclosure is about 2.6 mol% of the LNP. In some embodiments, the ratio of an additional pegylated lipid (e.g., compounds of Formula (V)), when present, to a compound of the present disclosure ranges from about 100:1 to about 5:1, for example about 50:1 to about 10:1. In some embodiments this ratio is about 100:1, 50:1, 40:1, 30:1, 20:1, 15:1, 10:1 or 5:1. In certain embodiments, the lipid nanoparticle comprises at least one steroid. In certain embodiments, the at least one steroid comprises cholesterol. In some embodiments, the molar ratio of the cationic lipid to cholesterol ranges from 5:1 to 1:1 or from 2:1 to 1:1 In certain embodiments, the steroid is cholesterol. In some embodiments, the steroid is present in the LNP at a concentration ranging from about 35 to about 45 mol%, or from about 41 to about 43 mol%. In some embodiments, the steroid is present in the LNP at a concentration of about 35 mol%. In some embodiments, the steroid is present in the LNP at a concentration of about 36 mol%. In some embodiments, the steroid is present in the LNP at a concentration of about 37 mol%. In some embodiments, the steroid is present in the LNP at a concentration of about 38 mol%. In some embodiments, the steroid is present in the LNP at a concentration of about 39 mol%. In some embodiments, the steroid is present in the LNP at a concentration of about 40 mol%. In some embodiments, the steroid is present in the LNP at a concentration of about 41 mol%. In some embodiments, the steroid is present in the LNP at a concentration of about 42 mol%. In some embodiments, the steroid is present in the LNP at a concentration of about 43 mol%. In some embodiments, the steroid is present in the LNP at a concentration of about 44 mol%. In some embodiments, the steroid is present in the LNP at a concentration of about 45 mol%. In various embodiments, the LNP or composition comprise about 40-50 mol% of the cationic lipid, about 5-15 mol% of the neutral lipid, about 35-45 mol% of the steroid, and about 0.05-0.20 mol% of the compound of the present disclosure, based on total mol of the lipids present in the lipid nanoparticle or composition. In various other embodiments, the LNP or composition comprise about 40-50 mol% of the cationic lipid, about 5-15 mol% of the neutral lipid, about 35-45 mol% of the steroid, about 2.0-3.0 mol% of a pegylated lipid, and about 0.05- 0.20 mol% of the compound of the present disclosure, based on total mol of the lipids present in the lipid nanoparticle or composition. In some embodiments, the LNP further comprises at least one therapeutic agent. In certain embodiments, the therapeutic agent comprises a nucleic acid. In some embodiments, the therapeutic agent is a nucleic acid. In certain embodiments, the nucleic acid comprises an antisense RNA, a messenger RNA, or a combination thereof. In some embodiments, the at least one therapeutic agent comprises Cas9 mRNA or ribonucleoprotein. In some embodiments, the messenger RNA encodes an antigen. In some embodiments, the antigen is an influenza antigen or a respiratory syncytial virus (RSV) antigen. In certain embodiments, the influenza antigen is an influenza A antigen or an influenza B antigen. In certain embodiments, the LNP has a size of about 40 nm to about 70 nm. In some embodiments, the lipid nanoparticle has a size of about 45 nm to about 65 nm, about 50 nm to about 60 nm, about 30 nm to about 70 nm, about 35 nm to about 75 nm, about 45 nm to about 80 nm, about 25 nm to about 100 nm, about 20 nm to about 90 nm, about 15 nm to about 150 nm, or about 10 nm to about 200 nm. One embodiment provides a pharmaceutical composition, comprising a LNP of the present disclosure and a pharmaceutically acceptable diluent or excipient. Administration of the compositions of the disclosure can be carried out via any of the accepted modes of administration of agents for serving similar utilities. The pharmaceutical compositions of the disclosure may be formulated into preparations in solid, semi-solid, liquid, or gaseous forms, such as tablets, capsules, powders, granules, ointments, solutions, suspensions, suppositories, injections, inhalants, gels, microspheres, and aerosols. Typical routes of administering such pharmaceutical compositions include, without limitation, oral, topical, transdermal, inhalation, parenteral, sublingual, buccal, rectal, vaginal, and intranasal. The term parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intradermal, intrasternal injection or infusion techniques. Pharmaceutical compositions of the disclosure are formulated to allow the active ingredients contained therein to be bioavailable upon administration of the composition to a patient. Methods of preparing such pharmaceutical compositions are known, or will be apparent, to those skilled in this art; for example, see Remington: The Science and Practice of Pharmacy, 20th Edition (Philadelphia College of Pharmacy and Science, 2000). The composition to be administered will, in any event, contain an LNP comprising a compound of the present disclosure and a therapeutically effective amount of a therapeutic agent for treatment of a disease or condition of interest in accordance with the teachings of this disclosure. A pharmaceutical composition of the disclosure may be in the form of a solid or liquid. In one aspect, the carrier(s) are particulate, so that the compositions are, for example, in tablet or powder form. The carrier(s) may be liquid, with the compositions being, for example, an oral syrup, injectable liquid, or an aerosol, which is useful in, for example, inhalatory administration. When intended for oral administration, the pharmaceutical composition is preferably in either solid or liquid form, where semi-solid, semi-liquid, suspension and gel forms are included within the forms considered herein as either solid or liquid. As a solid composition for oral administration, the pharmaceutical composition may be formulated into a powder, granule, compressed tablet, pill, capsule, chewing gum, wafer, or the like form. Such a solid composition will typically contain one or more inert diluents or edible carriers. In addition, one or more of the following may be present: binders such as carboxymethylcellulose, ethyl cellulose, microcrystalline cellulose, gum tragacanth or gelatin; excipients such as starch, lactose or dextrins, disintegrating agents such as alginic acid, sodium alginate, Primogel, corn starch and the like; lubricants such as magnesium stearate or Sterotex; glidants such as colloidal silicon dioxide; sweetening agents such as sucrose or saccharin; a flavoring agent such as peppermint, methyl salicylate or orange flavoring; and a coloring agent. When the pharmaceutical composition is in the form of a capsule, for example, a gelatin capsule, it may contain, in addition to materials of the above type, a liquid carrier such as polyethylene glycol or oil. The pharmaceutical composition may be in the form of a liquid, for example, an elixir, syrup, solution, emulsion, or suspension. The liquid may be for oral administration or for delivery by injection, as two examples. When intended for oral administration, preferred composition contain, in addition to the present LNPs, one or more of a sweetening agent, preservatives, dye / colorant and flavor enhancer. In a composition intended to be administered by injection, one or more of a surfactant, preservative, wetting agent, dispersing agent, suspending agent, buffer, stabilizer, and isotonic agent may be included. The liquid pharmaceutical compositions of the disclosure, whether they be solutions, suspensions or other like form, may include one or more of the following adjuvants: sterile diluents such as water for injection, saline solution, preferably physiological saline, Ringer's solution, isotonic sodium chloride, fixed oils such as synthetic mono or diglycerides which may serve as the solvent or suspending medium, polyethylene glycols, glycerin, propylene glycol or other solvents; antibacterial agents such as benzyl alcohol or methyl paraben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose; agents to act as cryoprotectants such as sucrose or trehalose. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic. Physiological saline is a preferred adjuvant. An injectable pharmaceutical composition is preferably sterile. A liquid pharmaceutical composition of the disclosure intended for either parenteral or oral administration should contain an amount of a LNP of the disclosure such that a suitable dosage of the therapeutic agent will be obtained. The pharmaceutical composition of the disclosure may be intended for topical administration, in which case the carrier may suitably comprise a solution, emulsion, ointment or gel base. The base, for example, may comprise one or more of the following: petrolatum, lanolin, polyethylene glycols, bee wax, mineral oil, diluents such as water and alcohol, and emulsifiers and stabilizers. Thickening agents may be present in a pharmaceutical composition for topical administration. If intended for transdermal administration, the composition may include a transdermal patch or iontophoresis device. The pharmaceutical composition of the disclosure may be intended for rectal administration, in the form, for example, of a suppository, which will melt in the rectum and release the drug. The composition for rectal administration may contain an oleaginous base as a suitable nonirritating excipient. Such bases include, without limitation, lanolin, cocoa butter, and polyethylene glycol. The pharmaceutical composition of the disclosure may include various materials, which modify the physical form of a solid or liquid dosage unit. For example, the composition may include materials that form a coating shell around the active ingredients. The materials that form the coating shell are typically inert, and may be selected from, for example, sugar, shellac, and other enteric coating agents. Alternatively, the active ingredients may be encased in a gelatin capsule. The pharmaceutical composition of the disclosure in solid or liquid form may include an agent that binds to the LNP of the disclosure and thereby assists in the delivery of the LNP. Suitable agents that may act in this capacity include a monoclonal or polyclonal antibody, or a protein. The pharmaceutical composition of the disclosure may consist of dosage units that can be administered as an aerosol. The term aerosol is used to denote a variety of systems ranging from those of colloidal nature to systems consisting of pressurized packages. Delivery may be by a liquefied or compressed gas or by a suitable pump system that dispenses the active ingredients. Aerosols of LNPs of the present disclosure may be delivered in single phase, bi-phasic, or tri-phasic systems to deliver the active ingredient(s). Delivery of the aerosol includes the necessary container, activators, valves, sub-containers, and the like, which together may form a kit. One skilled in the art, without undue experimentation may determine preferred aerosols. The pharmaceutical compositions of the disclosure may be prepared by methodology well known in the pharmaceutical art. For example, a pharmaceutical composition intended to be administered by injection can be prepared by combining the LNPs of the disclosure with sterile, distilled water or other carrier to form a solution. A surfactant may be added to facilitate the formation of a homogeneous solution or suspension. Surfactants are compounds that non-covalently interact with the compound of the present disclosure to facilitate dissolution or homogeneous suspension of the compound in the aqueous delivery system. The compositions of the disclosure, or their pharmaceutically acceptable salts, are administered in a therapeutically effective amount, which will vary depending upon a variety of factors including the activity of the specific therapeutic agent employed; the metabolic stability and length of action of the therapeutic agent; the age, body weight, general health, sex, and diet of the patient; the mode and time of administration; the rate of excretion; the drug combination; the severity of the particular disorder or condition; and the subject undergoing therapy. Compositions of the disclosure may also be administered simultaneously with, prior to, or after administration of one or more other therapeutic agents. Such combination therapy includes administration of a single pharmaceutical dosage formulation of a composition of the disclosure and one or more additional active agents, as well as administration of the composition of the disclosure and each active agent in its own separate pharmaceutical dosage formulation. For example, a composition of the disclosure and the other active agent can be administered to the patient together in a single oral dosage composition such as a tablet or capsule, or each agent administered in separate oral dosage formulations. Where separate dosage formulations are used, the compounds of the present disclosure and one or more additional active agents can be administered at essentially the same time, i.e., concurrently, or at separately staggered times, i.e., sequentially; combination therapy is understood to include all these regimens. Preparation methods for the above compounds and compositions are described herein below and / or known in the art. It will be appreciated by those skilled in the art that in the process described herein the functional groups of intermediate compounds may need to be protected by suitable protecting groups. Such functional groups include hydroxy, amino, mercapto and carboxylic acid. Suitable protecting groups for hydroxy include trialkylsilyl or diarylalkylsilyl (for example, t- butyldimethylsilyl, t-butyldiphenylsilyl or trimethylsilyl), tetrahydropyranyl, benzyl, and the like. Suitable protecting groups for amino, amidino and guanidino include t-butoxycarbonyl, benzyloxycarbonyl, and the like. Suitable protecting groups for mercapto include -C(O)-R″ (where R″ is alkyl, aryl or arylalkyl), p-methoxybenzyl, trityl and the like. Suitable protecting groups for carboxylic acid include alkyl, aryl or arylalkyl esters. Protecting groups may be added or removed in accordance with standard techniques, which are known to one skilled in the art and as described herein. The use of protecting groups is described in detail in Green, T.W. and P.G.M. Wutz, Protective Groups in Organic Synthesis (1999), 3rdEd., Wiley. As one of skill in the art would appreciate, the protecting group may also be a polymer resin such as a Wang resin, Rink resin or a 2-chlorotrityl-chloride resin. Furthermore, all compounds of this disclosure which exist in free base or acid form can be converted to their pharmaceutically acceptable salts by treatment with the appropriate inorganic or organic base or acid by methods known to one skilled in the art. Salts of the compounds of the present disclosure can be converted to their free base or acid form by standard techniques. SYNTHETIC EXAMPLE 1 PREPARATION OFGALNACPEGLIPIDCOMPOUND1-1 Synthesis of tert-butyl−Linker−PEGz−COOH (Compound 1-1D) Tert-butyl hydrogendodecanedioate (Cmpd.1-1A, 158 mg, 0.55 mmol, 1.0 eq) was dissolved in dry DCM (20 ml; 8 mg / ml). EDC.HCL (157 mg, 0.82 mmol, 1.5 eq) and NHS (76 mg, 0.66 mmol, 1.2 eq) were added to the solution and stirred at RT for 2 h to form Compound 1-1B. Then, PEG (Cmpd.1-1C, 1.1 g, 0.55 mmol, 1.0 eq) and triethylamine (114 μL, 0.82 mmol, 1.5 eq) were added to the reaction mixture and the resulting solution was stirred overnight at RT. The reaction mixture was washed with aqueous citric acid (3 × 50 mL, 0.1 M, pH 6). The separated organic phase was concentrated on the rotary evaporator. The crude product was dissolved in DCM and purified by plug filtration (20 g silica, DCM:MeOH 9:1). Tert- butyl−Linker−PEGz−COOH (Cmpd.1-1D, 1.2632 g, 0.56 mmol, quantitative yield) was obtained as a waxy, colorless solid.

[0013] Synthesis of tert-butyl−Linker−PEGz−CO−NHS (Compound 1-1E) Tert-Butyl-Linker-PEG-COOH (Cmpd.1-1D, 1.263 g, 0.56 mmol, 1 eq) was dissolved in dry DCM (60 mL; 21 mg / mL). DCC (229 mg, 1.11 mmol, 2 eq) and NHS (128 mg, 1.11 mmol, 2 eq) were added to the solution and stirred at RT for 2 h. Then, the reaction mixture was concentrated, and the residual crude product was dissolved in toluene (40 ml). Filtration removed any non-dissolved solid (dicyclohexylurea). To the filtrate, TBME (30 mL) was added and the solution was cooled to 0 °C. The resulting precipitate was collected via filtration. Drying the filter cake under vacuum gave the NHS ester (Cmpd.1-1E, 1.044 g, 0.44 mmol, 79% yield) as a white solid.

[0014] Synthesis of tert-butyl−Linker−PEG−Lipid (Compound 1-1G) Tert-butyl−Linker−PEG−CO−NHS (Cmpd.1-1E, 918 mg, 0.39 mmol, 1 eq) was dissolved in toluene (40 ml; 23 mg / mL) at 30°C. To this solution, dioctadecylamine (Cmpd.1- 1F, 221 mg, 0.43 mmol, 1.1 eq) and triethylamine (200 μL) was added and resulting reaction mixture was stirred overnight at 30 °C. Then, the solvent was removed, and the residual crude product was purified by plug filtration (20 g silica, DCM:MeOH 9:1) to give tert- Butyl−Linker−PEG−Lipid (Cmpd.1-1G, 618 mg, 0.22 mmol, 56% yield) as a waxy, white solid.

[0015] Synthesis of HOOC-Linker-PEG-Lipid (Compound 1-1H) To a solution of tert-Butyl-Linker-PEG-Lipid (Cmpd.1-1G, 570 mg, 0.24 mmol, 1 eq) in dry DCM (60 mL; 9.5 mg / ml) under N2 atmosphere, 10 volume % TFA (6 ml) was added and the formed reaction mixture was stirred at RT for 2 h. Then, the solvent was removed. Chromatographic purification of the residual crude product on silica (30g silica, DCM:MeOH 9:1), followed by MPLC purification on a RP-18 cartridge gave HOOC-Linker-PEG-Lipid (Cmpd.1-1H, 400 mg, 0.15 mmol, 61% yield) as a colorless, viscous oil. Synthesis of Compound 1-1J To a solution of HOOC-Linker-PEG-Lipid (Cmpd.1-1H, 535 mg, 0.2 mmol, 1.0 eq.) in a mixture of DCM (10 mL) and DMF (4 mL), tri-arm GalNAc-NH2 (commercially purchased Cmpd.1-1I, 358 mg, 0.2 mmol, 1.0 eq.) and HATU (152 mg, 0.4 mmol, 2.0 eq.) were added. Then, DIPEA (136 μL, 0.8 mmol, 4.0 eq.) was added and the reaction mixture was stirred overnight at 20-25°C. Then, the solvent was removed under reduced pressure. The obtained slightly red residue was dissolved in MeOH and the mixture was purified by MPLC using a Chromabond Flash RS40 C18 ec, 15-40 μm cartridge. Compound 1-1J (479 mg, 0.1 mmol, 53% yield) was obtained as a colorless solid.

[0016] Synthesis of tri-GalNAc−Linker−PEGz−Lipid (Compound 1-1) Compound 1-1J (479 mg, 0.11 mmol, 1 eq.) was dissolved in a mixture of MeOH and DCM (2:1, 30 mL, 16 mg / mL). Then, a 0.5 M MeONa solution (2.1 ml, 10 eq. of MeONa) was added to the reaction mixture and stirred at RT for 2h. The solvent was removed under reduced pressure at the rotary evaporator, and the crude product was purified by MPLC (MeOH / ACN, Chromabond Flash RS40 C18 ec cartridge, 15-40 μm). Compound 1 (130 mg, 30 % yield) was obtained as a colorless solid. EXAMPLE 1 LIPID NANOPARTICLE FORMATION Cationic lipid, DSPC, cholesterol, pegylated lipid, and GalNAc-pegylated lipid (compounds of the present invention and comparative GalNAc-pegylated lipid) were solubilized in ethanol at desired molar ratios, such that the total pegylated lipid content was maintained at 2.5 mol% (e.g., 47.5:10:40:2.425:0.075 or 47.5:10:40:2.35: 0.15). Lipid nanoparticles (LNP) were prepared at a total lipid to mRNA weight ratio of approximately 10:1 to 40:1. The mRNA (CH65 IgG, RNA Technologies, Montreal, Quebec) was diluted to 0.2 mg / mL in 10 to 50 mM citrate buffer, pH 4 to 6 or 10 to 25 mM acetate buffer, pH 4 to 6. Syringe pumps were used to mix the ethanolic lipid solution with the mRNA aqueous solution at a ratio of about 1:5 to 1:3 (vol / vol) with total flow rates above 15 mL / min. The ethanol was then removed, and the external buffer replaced with PBS by dialysis. Finally, the lipid nanoparticles were filtered through a 0.2 μm pore sterile filter. Lipid nanoparticle particle size was determined using quasi-elastic light scattering via a Nicomp 370 submicron particle sizer (Santa Barbara, CA). Alternatively, particle size can also be as determined by quasi-elastic light scattering using a Malvern Zetasizer Nano ZS (Malvern, UK). EXAMPLE 2 SERUM IGG EXPRESSION Studies were performed in 10-11-week-old C57BL / 6J, B6.129P2-Apoetm1Unc / J and B6.129S7-Ldlrtm1Her / J mice (Charles River or Inotiv) according to guidelines established by an institutional animal care committee (ACC) and the Canadian Council on Animal Care (CCAC). A single dose of mRNA-lipid nanoparticle (0.3 mg mRNA / kg) was systemically administered by tail vein injection and whole blood was collected at a specific time point (e.g., 24 and 48 hours) post-administration. The serum was subsequently separated by centrifuging the tubes of whole blood at 2000 × g for 10 minutes at 4 °C and stored at -80 °C until use for analysis. For immunoglobulin G (IgG) ELISA (Life Diagnostics Human IgG ELISA kit), the serum samples were diluted 2000-4000 fold with 1× diluent solution.100 µL of diluted serum was dispensed into an anti-human IgG coated 96-well plate in duplicate alongside human IgG standards and incubated in a plate shaker at 150 rpm at 25 °C for 45 minutes. The wells were washed 5 times with 1× wash solution using a plate washer (400 µL / well).100 µL of HRP conjugate was added into each well and incubated in a plate shaker at the same condition above. The wells were washed 5 times again with 1× wash solution using a plate washer (400 µL / well). 100 µL of TMB reagent was added into each well and incubated in a plate shaker at the same condition above. The reaction was stopped by adding 100 µL of Stop solution to each well. The absorbance was read at 450 nm (A450) with a microplate reader. The amount of human IgG in mouse serum was determined by plotting A450 values for the assay standard against human IgG concentration and applying a 4-parameter logistic fit. EXAMPLE 3 IN VIVO EVALUATION IN WT, APOE KO AND LDLR KO MICE Female wild type C57BL / 6J, ApoE knockout, and LDLR knockout mice (10-11 weeks of age) were dosed i.v. via tail vein with 0.3 mg / kg mRNA formulated in LNPs comprising cationic lipid of formula II as described in EXAMPLE 1. The activity was determined by measuring the amount of human IgG in mouse serum 24 and 48 hours after administration as described in EXAMPLE 2. Table 6: Formulation Characterization *Compound 2 (a comparative GalNAc-pegylated lipid) Table 7: Serum IgG Expression in WT, ApoE KO and LDLR KO

[0017] As indicated in Table 7, GalNAc LNPs exhibited enhanced activity in both ApoE and LDLR KO mice. Among these, LNP with Compound 1-1 demonstrated superior activity, with approximately 20% improvement compared to LNP with Compound 2 in the knockout (KO) mice. In LDLR KO mice, the administration of 0.15% Compound 1-1 LNP resulted in 66% of the activity observed with No GalNAc LNP in WT mice. In an additional study female wild type and LDLR knockout mice (8-9 weeks of age) were dosed i.v. via tail vein with 0.3 or 0.5 mg / kg mRNA formulated in LNPs comprising cationic lipid of formula II or III as described in EXAMPLE 1. The activity was determined by measuring the amount of human IgG in mouse serum 24 hours after administration as described in EXAMPLE 2. Table 8: Formulation Characterization: *Compound 3 (a comparative GalNAc-pegylated lipid) See also, e.g., Figures 3 and 4. EXAMPLE 4 SERUM IGG EXPRESSION IN MICE Female CD-1 mice (5 weeks of age) were dosed i.v. via tail vein with 0.15 or 0.3 or 0.6 mg / kg mRNA formulated in LNPs comprising cationic lipid of formula IV as described in EXAMPLE 1. Cardiac puncture was performed 24 hours post dose to collect serum for analysis. The activity was determined by measuring the amount of human IgG in mouse serum 24 hours after administration as described in EXAMPLE 2. Table 9: Formulation Characterization: See also, e.g., Figures 5A, 5B, and 5C. EXAMPLE 5 SERUM IGG EXPRESSION IN NON-HUMAN PRIMATES Studies were performed in 4.5-5.5-year-old non-naïve cynomologus macaques (Guangxi Guidong Promate Laboratory Animal Development Co., Ltd or Hainan New Source Biotech Co., Ltd) according to guidelines established by an institutional animal care committee (ACC) and current International Conference on Harmonization (ICS) Harmonized Tripartite Guidelines. A single dose of mRNA-lipid nanoparticle (0.5 or 1.5 mg mRNA / kg) was administered by a 1-hour intravenous infusion via a temporary catheter inserted in a peripheral vein and whole blood was collected at specified time points into K3EDTA tubes (between 1 hour and 168 hours) post- administration. The plasma was subsequently separated by centrifuging the tubes of whole blood at 3000 × g for 10 minutes at 2-8°C within one hour of collection and stored at -80 °C until use for analysis. For immunoglobulin G (IgG) ELISA (Life Diagnostics Human IgG ELISA kit), the serum samples were diluted 10-10000-fold with 1× diluent solution.100 µL of diluted serum was dispensed into an anti-human IgG coated 96-well plate in duplicate alongside human IgG standards and incubated in a plate shaker at 150 rpm at 25 °C for 45 minutes. The wells were washed 5 times with 1× wash solution using a plate washer (400 µL / well).100 µL of HRP conjugate was added into each well and incubated in a plate shaker at the same condition above. The wells were washed 5 times again with 1× wash solution using a plate washer (400 µL / well). 100 µL of TMB reagent was added into each well and incubated in a plate shaker at the same condition above. The reaction was stopped by adding 100 µL of Stop solution to each well. The absorbance is read at 450 nm (A450) with a microplate reader. The amount of human IgG in monkey plasma was determined by plotting A450 values for the assay standard against human IgG concentration and applying a 4-parameter logistic fit. This comparison was conducted at a dose of 0.5 or 1.5 mg mRNA / kg and expressed as µg IgG / mL serum, measured pre-dose, 6, 24, 48, 72, and 168 hours after administration (end of infusion). IgG peak plasma levels (Cmax) and area under the curve (AUC) were calculated from the IgG concentration profile measured over 7 days. Figures 2A and 2B provide plasma IgG1 levels for animals treated with: a) LNP comprising a cationic lipid of general formula III with and without GalNAc lipid 1-1 (Figure 2A; 0.5 mg / kg and 1.5 mg / kg); and b) LNP comprising a cationic lipid of general formula IV with and without GalNAc lipid 1-1 (Figure 2B; 0.5 mg / kg and 1.5 mg / kg). The data shows enhanced IgG1 expression in LNPs incorporating GalNAc lipid 1-1. At higher doses, the addition of GalNAc lipid 1-1 seems to reduce inter-animal variability. See also, e.g., Figure 6. Figure 6 shows plasma IgG levels for animals treated with LNP comprising a cationic lipid of general Formula II with and without GalNAc lipid 1-1 at doses of 0.5 mg / kg and 1.5 mg / kg. EXAMPLE 6 GALNAC TITRATION Non-naïve female cynomolgus monkeys (2.6–4.8 kg; 5.2–6.1 years old) were administered a single intravenous infusion of LNP-formulated mRNA encoding an IgG (CH65 2R.008) at a dose of 1.5 mg / kg. The LNPs comprised a cationic lipid, DSPC, cholesterol, PEG- lipid, and a GalNAc-PEG-lipid (either a compound of the present invention or a comparative compound) at varying molar percentages. Table 10: Formulation Characterization: See also, e.g., Figure 7A and 7B. The various embodiments described above can be combined to provide further embodiments. All of the U.S. patents, U.S. patent application publications, U.S. patent applications, foreign patents, foreign patent applications and non-patent publications referred to in this specification and / or listed in the Application Data Sheet, including U.S. Provisional Patent App. No.63 / 649,223, filed on May 17, 2024, and U.S. Provisional Patent App. No.63 / 707,658, filed on October 15, 2024, are incorporated herein by reference, in their entirety. Aspects of the embodiments can be modified, if necessary to employ concepts of the various patents, applications and publications to provide yet further embodiments. These and other changes can be made to the embodiments in light of the above-detailed description. In general, in the following claims, the terms used should not be construed to limit the claims to the specific embodiments disclosed in the specification and the claims, but should be construed to include all possible embodiments along with the full scope of equivalents to which such claims are entitled. Accordingly, the claims are not limited by the disclosure.

Claims

CLAIMS 1. A compound having the structure of Formula (I):or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof; wherein: R1, R2, and R3are each, independentlyL1, L2, L3, and L4are, at each occurrence, independently −NH(C=O)− or –(C=O)NH−; a is an integer from 4 to 12; b and c are each independently an integer from 1 to 12; d is an integer from 1 to 10; z has a mean value ranging from 1 to 60; and R4and R5are each independently a straight or branched alkyl, alkenyl or alkynyl chain containing from 10 to 30 carbon atoms, wherein the alkyl, alkenyl or alkynyl chain is optionally interrupted by one or more ester bonds.

2. The compound of claim 1, or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof, wherein L1is −NH(C=O)−.

3. The compound of claim 1 or 2, or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof, wherein L2is –(C=O)NH−.

4. The compound of any one of claims 1–3, or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof, wherein L3, at each occurrence, is –(C=O)NH−.

5. The compound of any one of claims 1–4, or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof, wherein L4, at each occurrence, is –NH(C=O)−.

6. The compound of any one of claims 1–5, having the structure of Formula (IA):or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof; wherein: R1, R2, and R3are each, independentlya is an integer from 6 to 10; b and c are each independently an integer from 1 to 6; d is an integer from 1 to 5; z has a mean value ranging from 1 to 60; and R4and R5are each independently a straight or branched alkyl, alkenyl or alkynyl chain containing from 10 to 30 carbon atoms, wherein the alkyl, alkenyl or alkynyl chain is optionally interrupted by one or more ester bonds.

7. The compound of any one of claims 1–6, or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof, wherein a is 8.

8. The compound of any one of claims 1–7, or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof, wherein b and c are, at each occurrence, independently 3 or 4.

9. The compound of any one of claims 1–8, or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof, wherein d is 1.

10. The compound of any one of claims 1–9, or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof, wherein R4and R5are each independently straight alkyl chains containing from 12 to 18 carbon atoms.

11. The compound of any one of claims 1–10, wherein the compound has one of the; ;; oror a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof.

12. The compound of any one of claims 1–11, or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof; wherein z has a mean value ranging from 15 to 60, 30 to 60 or 40 to 50.

13. The compound of any one of claims 1–11, or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof; wherein z spans a range that is selected such that the PEG portion of (I) or (II) has an average molecular weight of about 400 to about 6000 g / mol.

14. The compound of any one of claims 1–11, or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof, wherein z has a mean value of about 40-50 or 44-48.

15. A compound having the following structure:or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof.

16. A lipid nanoparticle comprising: the compound of any one of claims 1–15, or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof; and a therapeutic agent.

17. A composition comprising: the compound of any one of claims 1–15, or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof; and a therapeutic agent.

18. The lipid nanoparticle or composition of claim 16 or 17, further comprising one or more excipient selected from a neutral lipid, a steroid, and a cationic lipid.

19. The lipid nanoparticle or composition of any one of claims 16-18, comprising one or more neutral lipid selected from DSPC, DPPC, DMPC, DOPC, POPC, DOPE, and SM.

20. The lipid nanoparticle or composition of claim 18 or 19, wherein the neutral lipid is DSPC.

21. The lipid nanoparticle or composition of any one of claims 18-20, wherein the molar ratio of the cationic lipid to the neutral lipid ranges from about 2:1 to 8:

1.

22. The lipid nanoparticle or composition of any one of claims 18-21, wherein the steroid is cholesterol.

23. The lipid nanoparticle or composition of claim 22, wherein the molar ratio of the cationic lipid to cholesterol ranges from 5:1 to 1:1 or from 2:1 to 1:

1.

24. The lipid nanoparticle or composition of any one of claims 18-23, wherein the molar ratio of the cationic lipid to the compound of any one of claims 1-15 ranges from about 1000:1 to about 100:1 or from about 700:1 to about 200:1.

25. The lipid nanoparticle or composition of any one of claims 18-24, wherein theIIA IIB or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof, wherein: L21and L22are each independently -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O)x-, −S−S−, −C(=O)S−, −SC(=O)−, −NRaC(=O)−, −C(=O)NRa−, −NRaC(=O)NRa−, −OC(=O)NRa− or −NRaC(=O)O−; G23is C1-C6 alkylene; Rais H or C1-C12 alkyl; R21aand R21bare, at each occurrence, independently either: (a) H or C1-C12 alkyl; or (b) R21ais H or C1-C12 alkyl, and R21btogether with the carbon atom to which it is bound is taken together with an adjacent R21band the carbon atom to which it is bound to form a carbon-carbon double bond; R22aand R22bare, at each occurrence, independently either: (a) H or C1-C12 alkyl; or (b) R22ais H or C1-C12 alkyl, and R22btogether with the carbon atom to which it is bound is taken together with an adjacent R22band the carbon atom to which it is bound to form a carbon-carbon double bond; R23aand R23bare, at each occurrence, independently either (a): H or C1-C12 alkyl; or (b) R23ais H or C1-C12alkyl, and R23btogether with the carbon atom to which it is bound is taken together with an adjacent R23band the carbon atom to which it is bound to form a carbon-carbon double bond; R24aand R24bare, at each occurrence, independently either: (a) H or C1-C12 alkyl; or (b) R24ais H or C1-C12alkyl, and R24btogether with the carbon atom to which it is bound is taken together with an adjacent R24band the carbon atom to which it is bound to form a carbon-carbon double bond; R25and R26are each independently H or methyl;R27is C6-C16 alkyl; R28and R29are each independently C1-C12 alkyl; or R28and R29, together with the nitrogen atom to which they are attached, form a 5, 6 or 7-membered heterocyclic ring; e′, f′, g′, and h′ are each independently an integer from 1 to 24; and x is 0, 1 or 2.

26. The lipid nano particle or composition of claim 25, wherein the cationic lipid has one of the following structures:.

27. The lipid nanoparticle or composition of any one of claims 18-24, wherein the cationic lipid has the structure of Formula (III):or a pharmaceutically acceptable salt, tautomer, prodrug or stereoisomer thereof, wherein: one of L31or L32is –O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O)x-, -S-S-, -C(=O)S-, SC(=O)-, -NRaC(=O)-, -C(=O)NRa-, NRaC(=O)NRa-, -OC(=O)NRa- or -NRaC(=O)O-, and the other of L1or L2is –O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O)x-, -S-S-, -C(=O)S-, SC(=O)-, -NRaC(=O)-, -C(=O)NRa-, ,NRaC(=O)NRa-, -OC(=O)NRa- or -NRaC(=O)O- or a direct bond;G1and G2are each independently unsubstituted C1-C12 alkylene or C1-C12 alkenylene; G3is C1-C24 alkylene, C1-C24 alkenylene, C3-C8 cycloalkylene, C3-C8 cycloalkenylene; Rais H or C1-C12 alkyl; R31and R32are each independently C6-C24 alkyl or C6-C24 alkenyl; R33is H, OR35, CN, -C(=O)OR34, -OC(=O)R34or –NR35C(=O)R34; R34is C1-C12 alkyl; R35is H or C1-C6 alkyl; and x is 0, 1 or 2.

28. The lipid nanoparticle or composition of any one of claims 18-24, wherein the cationic lipid has one of the following structures:

29. The lipid nanoparticle or composition of any one of claims 18-24, wherein the cationic lipid has the structure of Formula (IV):or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof, wherein: R41is optionally substituted C1-C24 alkyl or optionally substituted C2-C24 alkenyl; R42and R43are each independently optionally substituted C1-C36 alkyl;R44and R45are each independently optionally substituted C1-C6 alkyl, or R44and R45join, along with the N to which they are attached, to form a heterocyclyl or heteroaryl; L41, L42, and L43are each independently optionally substituted C1-C18 alkylene; G41is a direct bond, -(CH2)nO(C=O)-, -(CH2)n(C=O)O-, or –(C=O)-; G42and G43are each independently -(C=O)O- or -O(C=O)-; and n is an integer greater than 0.

30. The lipid nanoparticle or composition of any one of claims 18-24, wherein the cationic lipid has one of the following structures:.

31. The lipid nanoparticle or composition of any one of claims 16-30 further comprising an additional pegylated lipid.

32. The lipid nanoparticle or composition of claim 31, wherein the additional pegylated lipid has the following Formula (III):or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof, wherein:R31and R32are each independently a straight or branched alkyl, alkenyl, or alkynyl containing from 10 to 30 carbon atoms, wherein each alkyl, alkenyl, or alkynyl is optionally substituted with at least one fluoro; and z′ is an integer ranging from 30 to 60.

33. The lipid nanoparticle or composition of claim 32, wherein R31and R32are each independently a straight alkyl chain containing from 12 to 16 carbon atoms.

34. The lipid nanoparticle or composition of claim 32 or 33, wherein z′ ranges from 45 to 50.

35. The lipid nanoparticle or composition of any one of claims 16-32, wherein the lipid nanoparticle or composition comprises about 40-50 mol% of the cationic lipid, about 5-15 mol% of the neutral lipid, about 35-45 mol% of the steroid, and about 0.05-0.20 mol% of the compound of any one of claims 1-15, based on total mol of the lipids present in the lipid nanoparticle or composition.

36. The lipid nanoparticle or composition of any one of claims 16-35, wherein the therapeutic agent comprises a nucleic acid.

37. The lipid nanoparticle or composition of claim 36, wherein the nucleic acid is an antisense RNA or a mRNA.

38. The lipid nanoparticle or composition of claim 37, wherein the mRNA encodes an antigen.

39. The lipid nanoparticle or composition of any one of claims 16-35, wherein the therapeutic agent comprises a Cas9 mRNA or a ribonucleoprotein.

40. The lipid nanoparticle or composition of any one of claims 16-39, further comprising from 2.0-2.5% of an additional pegylated lipid, preferably about 2.5%, and / or preferably wherein the additional pegylated lipid is a compound of any one of claims 28-30.

41. A pharmaceutical composition comprising the lipid nanoparticle or composition of any one of claims 16-40 and one or more pharmaceutically acceptable excipients.

42. A method for inducing expression of a desired protein in a subject in need thereof, the method comprising administering a therapeutically effective amount of the pharmaceutical composition of claim 41 to the subject, wherein the therapeutic agent comprises a nucleic acid.

43. The method of claim 42, wherein the nucleic acid is an antisense RNA, a mRNA or a Cas9 mRNA.

44. The method of claim 43, wherein mRNA encodes an antigen.

45. The method of any one of claims 42-44, wherein the method is for vaccination against a viral pathogen.

46. The method of claim 43, wherein the nucleic acid is a Cas9 mRNA, and the method is for editing a gene of interest.

Images