Lipid nanoparticles comprising coding RNA molecules for use in gene editing and as vaccines and therapeutic agents

Abstract

The present disclosure describes improved LNP-based RNA vaccines, nucleobase editing systems, and therapeutics for use in treating and / or immunization against disease. In particular, the disclosure describes improved LNPs, including novel and improved ionizable lipids for making LNPs, that enhance the targeted delivery of LNP-based RNA vaccines and therapeutics based on linear and / or circular mRNAs. The improved LNPs protect linear and / or circular mRNA payloads from degradation and clearance while achieving targeted systemic or local delivery for use as enhanced vaccines and / or therapeutic agents.

Description

LIPID NANOPARTICLES COMPRISING CODING RNA MOLECULES FOR USE IN GENE EDITING AND AS VACCINES AND THERAPEUTIC AGENTS TECHNICAL FIELD

[0001] The present disclosure generally relates to the field of nucleic acid lipid nanoparticle (LNP) compositions and delivery thereof for use as vaccines and / or therapeutics for the treatment of disease. The disclosure further relates to compositions comprising LNPs formulated with coding RNAs, including linear and / or circular mRNAs, for the delivery of encoded vaccine antigens and / or therapeutic proteins for the vaccination against infectious agents and / or treatment of disease, including infectious disease and cancer. BACKGROUND

[0002] There are many challenges associated with the delivery of nucleic acids to affect a desired response in a biological system, such as an immune response or the production of a therapeutically beneficial protein to treat a disease. Nucleic acid-based therapeutics and vaccines have enormous potential but there remains a need for more effective delivery of nucleic acids to appropriate sites within a cell or organism in order to realize this potential.

[0003] Nucleic acid-based therapeutics and vaccines are generally composed of DNA or RNA. DNA is known to be relatively stable and easy to handle, however, the use of DNA bears the risk of undesired insertion into a cell’s genome which potentially may produce mutagenic events. As a further concern, the delivery of DNA is associated with unwanted immunogenicity and the production of anti-DNA antibodies. Yet another concern in the use of DNA is the limited expression level of the encoded peptide or protein that is achievable due to the requirement that the administered DNA must first enter the nucleus to undergo transcription prior to translation into a desired protein product (e.g., antigen or therapeutic protein).

[0004] In contrast to DNA, the use of RNA is substantially safer because RNA does not involve the risk of being integrated into the genome of a transfected cell, thus eliminating the concern that the introduced genetic material will disrupt the normal functioning of an essential gene or cause a mutation. In addition, RNA-based agents do not require extraneous promoter sequences for effective expression of an encoded protein and are also less immunogenic than DNA-based agents, in part because RNA has a relatively short half-life unlike DNA. In addition, while DNA must enter the nuclease to perform its function, RNA performs its function outside of the nucleus and is therefore more efficient.

[0005] Despite the advantages of using RNA-based therapeutics and vaccines, the stability of RNA (e.g., mRNA) is far lower than DNA, especially when it reaches the cytoplasm of a cell and is exposed to RNA-degrading enzymes. In addition, the presence of a hydroxyl group on the second carbon of the sugar moiety in RNA causes steric hindrance that prevents the RNA from forming a more stable double helix structure like in the case of DNA, thus making RNA more prone to hydrolytic degradation than DNA.

[0006] To circumvent these challenges, delivery of RNA vaccines (e.g., mRNA vaccines) and therapeutics has recently focused on the use of lipid nanoparticles (LNPs). Indeed, LNPs have emerged as the most promising nonviral delivery vehicle for exogenous mRNA (see e.g., Guan et al., “Nanotechnologies in delivery of mRNA therapeutics using nonviral vector-based delivery systems,” Gene Ther, 24 (2017), pp.133-143). The LNP is a complex nanostructured body that provides protection to payload RNA molecules encapsulated within from the harshly degrading nuclease environment in vivo while facilitating intracellular delivery. LNPs are formed through self-assembly by combining the RNA payload with several lipid components, including an ionizable lipid that plays a central role in delivery efficacy (e.g., Miao et al., “Delivery of mRNA vaccines with heterocyclic lipids increases anti-tumor efficacy by STING-mediated immune cell activation,” Nat. Biotechnol., 27 (2019), pp.1174-1185). Entrapment of RNA is achieved by mixing RNA with lipids at an acidic pH at which the ionizable lipid is positively charged, thus ensuring a charge-driven interaction with the negatively charged RNA molecules (e.g., Mindy et al., “Mechanism of macromolecular structure evolution in self-assembled lipid nanoparticles for siRNA delivery,” Langmuir, 20 (2014), pp.4613-4622). The pH is then adjusted to above the pKa of the ionizable lipid, which results in a near-neutral surface charge desirable for clinical administration (see Id.). In addition, the incorporation of a pegylated lipid into the mixture achieves a sterically stabilized core shell nanoparticle useful for clinical applications as vaccines and / or therapeutics.

[0007] In spite of the development of LNPs, the delivery of RNA payloads to cells in vivo in a targeted manner that also allows for sufficient levels of protein production (e.g., production of vaccine antigens or therapeutic proteins) remains an important and significant challenge.

[0008] Genome editing tools encompass a diverse set of technologies that can make many types of genomic alterations in various contexts. These technologies have evolved over the last couple of decades to provide a range of user-programmable editing tools that include ZFN (zinc finger) nuclease editing systems, meganuclease editing systems, and TALENS (transcription activator-like effector nucleases). The past decade has seen an explosive growth in a new generation of genome editing systems based on components from bacterial immune pathways, including CRISPR (clustered regularly interspaced short palindromic repeats) and the associated CRISPR-associated proteins (e.g., CRISPR-Cas9) (Jinek et al., “A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity,” Science, Vol.337 (6096), pp.816-821), meganuclease editors (Boissel et al., “megaTALs: a rare-cleaving nuclease architecture for therapeutic genome engineering,” Nucleic Acids Research 42: pp.2591-2601) and bacterial retron systems (Schubert et al., “High-throughput functional variant screens via in vivo production of single-stranded DNA,” PNAS, April 27, 2021, Vol.118(18), pp.1-10). In particular, CRISPR-Cas9 has been derivatized in numerous ways to expand upon its guide RNA-based programmable double-strand cutting activity to form systems ranging from finding alternative CRISPR Cas nuclease enzymes having different PAM requirements and cutting properties (e.g., engineered Cas9 proteins and other naturally-occurring Cas9 homologs,including, but not limited to, Cas12a, Cas12f, Cas13a, and Cas13b, and their engineered variants) to base editing (Komor et al., “Programmable editing of a target base in genomic DNA without double- stranded DNA cleavage,” Nature, May 19, 2016, 533 (7603); pp.420-424 [cytosine base editors or CBEs] and Gaudelli et al., “Programmable base editing of A-T to G-C in genomic DNA without DNA cleavage,” Nature, Vol.551, pp.464-471 [adenine base editors or ABEs]) to prime editing (Anzalone et al., “Search-and-replace genome editing without double-strand breaks or donor DNA,” Nature, Dec 2019, 576 (7789): pp.149-157) to twin prime editing (Anzalone et al., “Programmable deletion, replacement, integration and inversion of large DNA sequences with twin prime editing,” Nature Biotechnology, Dec 9, 2021, vol.40, pp.731-740) to epigenetic editing (Kungulovski and Jeltsch, “Epigenome Editing: State of the Art, Concepts, and Perspective,” Trends in Genetics, Vol.32, 206, pp.101-113) to CRISPR-directed integrase editing (Yarnell et al., “Drag-and-drop genome insertion of large sequences without double-stranded DNA cleavage using CRISPR-directed integrases,” Nature Biotechnology, Nov 24, 2022, (“PASTE”)).

[0009] While the expansion of genome editing tools has exploded, the development of safe and effective gene editing tool delivery systems has lagged behind. There remain numerous challenges associated with the delivery of gene editing tools—including, but not limited to, CRISPR- Cas9 and alternative Cas nuclease editors, retron editors, base editors, prime editors, twin prime editors, epigenetic editors, and integrase editors—to achieve safe and effective therapeutic application of such tools in cells and patients for treating disease and / or otherwise modifying the nucleotide sequence of a target nucleic acid molecule (e.g., a gene or genome) particularly as it relates to delivery in vivo. That said, the use of lipid nanoparticles (LNPs) has emerged as a leading delivery option for the safe, effective, and targeted delivery of gene editing tools to target tissues and cells. However, there remains a need for improved LNPs, including better performing ionizable lipids, that will enhance the targeted delivery of LNP-based gene editing tools. Preferably, such improved LNPs would protect payloads from degradation and clearance while achieving targeted delivery, be suitable for systemic or local delivery, and provide delivery of RNA cargo, including those relating to a wide variety of gene editing tools, such as those mentioned above. In addition, such improved LNP-based therapeutics should exhibit low toxicity and provide an adequate therapeutic index, such that patient treatment at an effective dose of the LNP minimizes risk to the patient while maximizing therapeutic benefit.

[0010] Thus, improved LNPs that enhance the delivery of LNP-based RNA vaccines and therapeutics to cells, tissues, and bodily sites and which are more protective of RNA payloads would advance the art. Preferably, such improved LNPs would protect RNA payloads from degradation and clearance while achieving delivery, be suitable for ex vivo or in vivo delivery , and provide delivery of any target, including RNA in linear and / or circular and / or modified form.. In addition, such improved LNP-based RNA vaccines and therapeutics should exhibit low toxicity and provide an adequate therapeutic index, such that patient treatment at an effective dose of the LNP minimizes risk to thepatient while maximizes therapeutic benefit. The present disclosure provides these and related advantages. SUMMARY

[0011] Described herein are compositions, methods, processes, kits and devices for the selection, design, preparation, manufacture, formulation, and / or use of LNP-based RNA medicines (e.g., vaccines and gene-editing therapeutics). In particular, described herein are compositions, methods, processes, kits and devices for the selection, design, preparation, manufacture, formulation, and / or use of LNP-based RNA medicines (e.g., vaccines and gene editing therapeutics) for the delivery of one or more coding and / or non-coding RNA molecules. In various embodiments, the non- coding RNAs may comprise one or more guide RNAs relating to a gene editing system, such as one based on CRISPR-Cas9 or CRISPR-Cas12a, each of which require complexing with a guide RNA that facilitates the localizing the protein-RNA complex to a target sequence having an enzyme- specific PAM site (protospacer adjacent motif – recognized by the CRISPR enzyme) and a target nucleotide sequence (i.e., the protospacer) that is complementary to a portion of the guide RNA (i.e., to the spacer region). In other embodiments, the coding RNA may encode any protein component of LNP-based RNA medicine, such as, but limited to a virus antigen (e.g., a viral envelope spike protein), a therapeutic protein (e.g., a functional version of a defective protein), or one or more gene editing components (e.g., a programmable nuclease or other effector protein, such as a deaminase or reverse transcriptase). Further described herein are compositions, methods, processes, kits and devices for the selection, design, preparation, manufacture, formulation, and / or use of LNP-based RNA medicines (e.g., vaccines and / or gene editing therapeutics) for the delivery of one or more RNA molecules, e.g., a coding RNA that codes for one or more therapeutic proteins for the prophylactic and / or therapeutic treatment of one or more diseases or a symptom thereof, or a non-coding RNA, such as, but not limited to a guide RNA for a gene editing system. In various embodiments, the RNA molecule delivered by the herein disclosed LNPs can be a linear mRNA. In other embodiments, the RNA molecule delivered by the herein disclosed LNPs can be a circular mRNA. In still other embodiments, the RNA molecule delivered by the herein disclosed LNPs can include both linear and circular forms of mRNA. In further embodiments, the RNA may comprise one or more modifications, including chemical modifications (e.g., ribonucleotide analogs, alternative phosphate chain linkers), sequence modification (e.g., relative to a wild type sequence), and / or structural modification (e.g., secondary-folded structures, such as, but not limited to, stem-loops, hairpins, and G-quadruplexes, and tertiary structural elements, such as, but not limited to, helical duplexes and triple-stranded structures). In various other embodiments, the disclosure provides novel lipid components of the herein disclosed LNPs, including, but not limited to, novel ionizable lipids.

[0012] The present disclosure describes improved LNP-based RNA medicines (e.g., vaccines and therapeutics) for use in treating and / or immunization against disease. In particular, the disclosure describes improved LNPs, including better performing ionizable lipids, that enhance the targeteddelivery of LNP-based RNA vaccines and therapeutics based on linear and / or circular mRNAs. The improved LNPs protect linear and / or circular mRNA cargos (i.e., the circular and / or linear mRNA molecules encapsulated by the LNPs) from degradation and clearance while achieving targeted systemic or local delivery for use as enhanced vaccines and / or therapeutic agents.

[0013] In an aspect of the disclosure, provided herein is a compound having a structure of any of Formulae (AX’’’), (AX’’), (AX’), (AX*), (AX), (AX-A), (AX-AZ), (AX-B), (AX-C), (AX-D), (AX-E), (AX-E’), (AX-F), (AX-G), (AX-G’), (AX-H), (AX-H’), (AX-H’’), (AX-H’’’), (AX-H’’’’), (AX-I), (AX-I’), (AX-I’’), (AX-I’’’), (AX-I’’’’), (AX-J), (AX-J’), (AX-J’’), (AX-J’’’), (AX-J’’’’), (AX-K), (AX-K’), (AX-K’’), (AX-K’’’), (AX-K’’’’), (AX-L), (AX-L’), (AX-L’’), (AX-M), (AX- M’), (AX-M’’), (AX-O), (AX-O’), (AX-O’’), (AX-P), (AX-P’), (AX-P’’), (AX-Q), (AX-Q’), (AX- Q’’), (AX-R), (AX-R’), (AX-R’’), (AX-S), (AX-S’), (AX-S’’), (AX-T), (AX-T’), (AX-T’’), (AX- U), (AX-U’), (AX-U’’), (AX-V), (AX-V’), (AX-V’’), (AX-W), (AX-W’), (AX-W’’), (AX-X), (AX- X’), (AX-X’’), (AX-Y) (AX-Y’), (AX-Y’’), (AX-Z), (AX-Z’), (AX-Z’’), (AX-AA), (AX-AA’), (AX- AA’’), (AX-AB), (AX-AB’), (AX-AB’’), (AX-AC), (AX-AC’), (AX-AC’’), (AX-AD), (AX-AD’), (AX-AD’’), (AX-AE), (AX-AE’), (AX-AE’’), (AX-AF), (AX-AF’), (AX-AF’’), (AX-AG), (AX- AG’), (AX-AG’’), (AX-AH), (AX-AH’), (AX-AH’’), (AX-AH’’’), (AX-AI), (AX-AI’), (AX-AI’’), (AX-AI’’’), (AX-AK), (AX-AL), (AX-AL’), (AX-AL’’), (AX-AL’’’), (AX-AM), (AX-AN), (AX- AO), (AX-AO’), (AN), (AN-A), (AN-B), and (AN-C) a pharmaceutically acceptable salt, solvate, stereoisomer, or enantiomer thereof, or any lipid in Table (I), or a salt, solvate, stereoisomer, or enantiomer thereof, see below, collectively referred to as "Lipids of the Disclosure" and each individually referred to as a "Lipid of the Disclosure."

[0014] In an aspect of the disclosure, provided herein is a pharmaceutical composition comprising a compound as disclosed herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0015] In an aspect, provided herein is a pharmaceutical composition comprising: a) at least one lipid nanoparticle comprising at least one compound having a structure of any of Formulae (AX’’’), (AX’’), (AX’), (AX*), (AX), (AX-A), (AX-AZ), (AX-B), (AX-C), (AX-D), (AX-E), (AX- E’), (AX-F), (AX-G), (AX-G’), (AX-H), (AX-H’), (AX-H’’), (AX-H’’’), (AX-H’’’’), (AX-I), (AX- I’), (AX-I’’), (AX-I’’’), (AX-I’’’’), (AX-J), (AX-J’), (AX-J’’), (AX-J’’’), (AX-J’’’’), (AX-K), (AX- K’), (AX-K’’), (AX-K’’’), (AX-K’’’’), (AX-L), (AX-L’), (AX-L’’), (AX-M), (AX-M’), (AX-M’’), (AX-O), (AX-O’), (AX-O’’), (AX-P), (AX-P’), (AX-P’’), (AX-Q), (AX-Q’), (AX-Q’’), (AX-R), (AX-R’), (AX-R’’), (AX-S), (AX-S’), (AX-S’’), (AX-T), (AX-T’), (AX-T’’), (AX-U), (AX-U’), (AX-U’’), (AX-V), (AX-V’), (AX-V’’), (AX-W), (AX-W’), (AX-W’’), (AX-X), (AX-X’), (AX-X’’), (AX-Y) (AX-Y’), (AX-Y’’), (AX-Z), (AX-Z’), (AX-Z’’), (AX-AA), (AX-AA’), (AX-AA’’), (AX- AB), (AX-AB’), (AX-AB’’), (AX-AC), (AX-AC’), (AX-AC’’), (AX-AD), (AX-AD’), (AX-AD’’), (AX-AE), (AX-AE’), (AX-AE’’), (AX-AF), (AX-AF’), (AX-AF’’), (AX-AG), (AX-AG’), (AX- AG’’), (AX-AH), (AX-AH’), (AX-AH’’), (AX-AH’’’), (AX-AI), (AX-AI’), (AX-AI’’), (AX-AI’’’),(AX-AK), (AX-AL), (AX-AL’), (AX-AL’’), (AX-AL’’’), (AX-AM), (AX-AN), (AX-AO), (AX- AO’), (AN), (AN-A), (AN-B), and (AN-C) or a pharmaceutically acceptable salt, solvate, stereoisomer, or enantiomer thereof, or any lipid in Table (I), or a salt, solvate, stereoisomer, or enantiomer thereof; and b) at least one nucleobase editing system.

[0016] In an aspect, provided herein is a method of delivering a nucleobase editing system to a subject in need thereof, the method comprising administering to the subject a pharmaceutical composition disclosed herein.

[0017] In an aspect, provided herein is a pharmaceutical composition comprising: a) at least one lipid nanoparticle comprising at least one compound having a structure of any of Formulae (AX’’’), (AX’’), (AX’), (AX*), (AX), (AX-A), (AX-AZ), (AX-B), (AX-C), (AX-D), (AX-E), (AX- E’), (AX-F), (AX-G), (AX-G’), (AX-H), (AX-H’), (AX-H’’), (AX-H’’’), (AX-H’’’’), (AX-I), (AX- I’), (AX-I’’), (AX-I’’’), (AX-I’’’’), (AX-J), (AX-J’), (AX-J’’), (AX-J’’’), (AX-J’’’’), (AX-K), (AX- K’), (AX-K’’), (AX-K’’’), (AX-K’’’’), (AX-L), (AX-L’), (AX-L’’), (AX-M), (AX-M’), (AX-M’’), (AX-O), (AX-O’), (AX-O’’), (AX-P), (AX-P’), (AX-P’’), (AX-Q), (AX-Q’), (AX-Q’’), (AX-R), (AX-R’), (AX-R’’), (AX-S), (AX-S’), (AX-S’’), (AX-T), (AX-T’), (AX-T’’), (AX-U), (AX-U’), (AX-U’’), (AX-V), (AX-V’), (AX-V’’), (AX-W), (AX-W’), (AX-W’’), (AX-X), (AX-X’), (AX-X’’), (AX-Y) (AX-Y’), (AX-Y’’), (AX-Z), (AX-Z’), (AX-Z’’), (AX-AA), (AX-AA’), (AX-AA’’), (AX- AB), (AX-AB’), (AX-AB’’), (AX-AC), (AX-AC’), (AX-AC’’), (AX-AD), (AX-AD’), (AX-AD’’), (AX-AE), (AX-AE’), (AX-AE’’), (AX-AF), (AX-AF’), (AX-AF’’), (AX-AG), (AX-AG’), (AX- AG’’), (AX-AH), (AX-AH’), (AX-AH’’), (AX-AH’’’), (AX-AI), (AX-AI’), (AX-AI’’), (AX-AI’’’), (AX-AK), (AX-AL), (AX-AL’), (AX-AL’’), (AX-AL’’’), (AX-AM), (AX-AN), (AX-AO), (AX- AO’), (AN), (AN-A), (AN-B), and (AN-C) or a pharmaceutically acceptable salt, solvate, stereoisomer, or enantiomer thereof, or any lipid in Table (I), or a salt, solvate, stereoisomer, or enantiomer thereof; and b) at least one mRNA or circRNA encoding a chimeric antigen protein.

[0018] In an aspect, provided herein is a method of generating a CAR T or CAR NK cell in a subject in need thereof, the method comprising administering to the subject a pharmaceutical composition disclosed herein.

[0019] In an aspect, provided herein is a pharmaceutical composition comprising: a) at least one lipid nanoparticle comprising at least one compound having a structure of any of Formulae (AX’’’), (AX’’), (AX’), (AX*), (AX), (AX-A), (AX-AZ), (AX-B), (AX-C), (AX-D), (AX-E), (AX- E’), (AX-F), (AX-G), (AX-G’), (AX-H), (AX-H’), (AX-H’’), (AX-H’’’), (AX-H’’’’), (AX-I), (AX- I’), (AX-I’’), (AX-I’’’), (AX-I’’’’), (AX-J), (AX-J’), (AX-J’’), (AX-J’’’), (AX-J’’’’), (AX-K), (AX- K’), (AX-K’’), (AX-K’’’), (AX-K’’’’), (AX-L), (AX-L’), (AX-L’’), (AX-M), (AX-M’), (AX-M’’), (AX-O), (AX-O’), (AX-O’’), (AX-P), (AX-P’), (AX-P’’), (AX-Q), (AX-Q’), (AX-Q’’), (AX-R), (AX-R’), (AX-R’’), (AX-S), (AX-S’), (AX-S’’), (AX-T), (AX-T’), (AX-T’’), (AX-U), (AX-U’), (AX-U’’), (AX-V), (AX-V’), (AX-V’’), (AX-W), (AX-W’), (AX-W’’), (AX-X), (AX-X’), (AX-X’’), (AX-Y) (AX-Y’), (AX-Y’’), (AX-Z), (AX-Z’), (AX-Z’’), (AX-AA), (AX-AA’), (AX-AA’’), (AX-AB), (AX-AB’), (AX-AB’’), (AX-AC), (AX-AC’), (AX-AC’’), (AX-AD), (AX-AD’), (AX-AD’’), (AX-AE), (AX-AE’), (AX-AE’’), (AX-AF), (AX-AF’), (AX-AF’’), (AX-AG), (AX-AG’), (AX- AG’’), (AX-AH), (AX-AH’), (AX-AH’’), (AX-AH’’’), (AX-AI), (AX-AI’), (AX-AI’’), (AX-AI’’’), (AX-AK), (AX-AL), (AX-AL’), (AX-AL’’), (AX-AL’’’), (AX-AM), (AX-AN), (AX-AO), (AX- AO’), (AN), (AN-A), (AN-B), and (AN-C) or a pharmaceutically acceptable salt, solvate, stereoisomer, or enantiomer thereof, or any lipid in Table (I), or a salt, solvate, stereoisomer, or enantiomer thereof; and b) at least one mRNA or circRNA encoding a gene editing system capable of executing an edit that treats a hemoglobinopathy.

[0020] In an aspect of the disclosure, provided herein is a lipid nanoparticle (LNP) comprising a compound having a structure of any of Formulae (AX’’’), (AX’’), (AX’), (AX*), (AX), (AX-A), (AX-AZ), (AX-B), (AX-C), (AX-D), (AX-E), (AX-E’), (AX-F), (AX-G), (AX-G’), (AX-H), (AX-H’), (AX-H’’), (AX-H’’’), (AX-H’’’’), (AX-I), (AX-I’), (AX-I’’), (AX-I’’’), (AX-I’’’’), (AX-J), (AX-J’), (AX-J’’), (AX-J’’’), (AX-J’’’’), (AX-K), (AX-K’), (AX-K’’), (AX-K’’’), (AX-K’’’’), (AX- L), (AX-L’), (AX-L’’), (AX-M), (AX-M’), (AX-M’’), (AX-O), (AX-O’), (AX-O’’), (AX-P), (AX- P’), (AX-P’’), (AX-Q), (AX-Q’), (AX-Q’’), (AX-R), (AX-R’), (AX-R’’), (AX-S), (AX-S’), (AX- S’’), (AX-T), (AX-T’), (AX-T’’), (AX-U), (AX-U’), (AX-U’’), (AX-V), (AX-V’), (AX-V’’), (AX- W), (AX-W’), (AX-W’’), (AX-X), (AX-X’), (AX-X’’), (AX-Y) (AX-Y’), (AX-Y’’), (AX-Z), (AX- Z’), (AX-Z’’), (AX-AA), (AX-AA’), (AX-AA’’), (AX-AB), (AX-AB’), (AX-AB’’), (AX-AC), (AX- AC’), (AX-AC’’), (AX-AD), (AX-AD’), (AX-AD’’), (AX-AE), (AX-AE’), (AX-AE’’), (AX-AF), (AX-AF’), (AX-AF’’), (AX-AG), (AX-AG’), (AX-AG’’), (AX-AH), (AX-AH’), (AX-AH’’), (AX- AH’’’), (AX-AI), (AX-AI’), (AX-AI’’), (AX-AI’’’), (AX-AK), (AX-AL), (AX-AL’), (AX-AL’’), (AX-AL’’’), (AX-AM), (AX-AN), (AX-AO), (AX-AO’), (AN), (AN-A), (AN-B), and (AN-C) or a pharmaceutically acceptable salt, solvate, stereoisomer, or enantiomer thereof, or any lipid in Table (I), or a salt, solvate, stereoisomer, or enantiomer thereof.

[0021] In another aspect of the disclosure, provided herein is a method for delivering a nucleic acid to a cell comprising contacting the cell with a LNP disclosed herein or a pharmaceutical composition disclosed herein.

[0022] In another aspect of the disclosure, provided herein is a method for treating a disease characterized by a deficiency of a functional protein, the method comprising administering to a subject having the disease, a LNP formulation comprising a LNP disclosed herein, wherein the mRNA encodes the functional protein or a protein having the same biological activity as the functional protein.

[0023] In another aspect of the disclosure, provided he rein is a method for treating a disease characterized by overexpression of a polypeptide, comprising administering to a subject having the disease a LNP formulation comprising a LNP disclosed herein and a siRNA, wherein the siRNA targets expression of the overexpressed polypeptide.BRIEF DESCRIPTION OF THE DRAWINGS

[0024] FIG.1 is a diagram illustrating the LNP-based RNA vaccines and therapeutics disclosed herein which are encapsulated with RNA payloads (e.g., linear and / or circular mRNAs).

[0025] FIG.2 is a diagram illustrating an originator polynucleotide construct of the present disclosure which may be linear or circular.

[0026] FIGs.3A-3C are diagrams illustrating exemplary chimeric antigen receptor (CAR) proteins of the present disclosure. FIG.3A shows representative CAR designs, showing the various constituent features of the CAR protein, wherein the pair of dashed lines represent the transmembrane region. FIGs.3B and 3C show representative humanized and ubiquitin-resistant variant CAR designs of the present disclosure.

[0027] FIG.4 is a graph showing the percentage of VHH+ total T cells, CD4+ T Cells and CD8+ T Cells isolated from non-human primate (NHP) spleen at 6, 24 and 48 hours after treatment with PBS or LNP formulation F-32 containing VHH mRNA. NHPs were dosed with test articles as described in Example 9A.

[0028] FIG.5A is a set of representative plots of non-human primate (NHP) T cells isolated from PBMCs at 6, 24 and 48 hours after treatment with PBS or LNP formulation F- 32c containing VHH mRNA, gated to measure VHH+ cells. NHPs were dosed with test articles as described in Example 9B.

[0029] FIG.5B is a set of graphs showing the percentage of VHH+ total T cells, CD4+ T Cells and CD8+ T Cells isolated from NHP PBMCs at 6, 24 and 48 hours after treatment with PBS or LNP formulation F-32c containing VHH mRNA. NHPs were dosed with test articles as described in Example 9B.

[0030] FIG.6A is a schematic of the experimental design of Example 10, showing human CD34+ HSC engraftment into NSG mice, dosing of test articles after 12 weeks, and collection of blood and relevant organs, including spleen.

[0031] FIG.6B is a set of representative flow cytometry plots of humanized mouse spleen cells after treatment with PBS or LNP formulations Control (fLuc control) and F-32a (VHH mRNA), gated to measure VHH+ cells in the total T cell population, CD4+ T cells and CD8+ T cells. Mice were dosed with test articles and samples were analyzed as described in Example 10.

[0032] FIG.6C is a graph showing the frequency of VHH reporter+ cells in the total T cell population, CD4+ T cells and CD8+ T cells in the spleen of humanized mice at 24 hrpost dosing. Mice were dosed with test articles and samples were analyzed as described in Example 10.

[0033] FIG.6D is a graph showing the frequency of VHH reporter+ cells in the total T cell population, CD4+ T cells and CD8+ T cells in a peripheral blood sample collected from humanized mice at 6 hr and 24 hr post dosing. Mice were dosed with test articles and samples were analyzed as described in Example 10.

[0034] FIG.7A is a set of representative flow cytometry plots of purified human T cells cultured in vitro in the presence of IL-2 (20 IU / mL) and TransAct before treatment with media alone, or with LNP formulation F-X3 containing anti-CD19-CAR mRNA Hz-828BBz.

[0035] FIG.7B is a bar graph plot of the data shown in the flow cytometry data in FIG.7A. Cells were cultured and analyzed as described in Example 15C.

[0036] FIGs.8A, 8B, 8C and 8D are graphs showing the results of Example 18, demonstrating B Cell depletion in humanized NSG mice. FIG.8A is a graph showing overall % of viable B cells in spleens 48 hr after dosing of formulations F-32 (VHH control) and F- X6 (containing anti-CD19-CAR construct m-283z). FIG.8B is a graph showing absolute count of viable B cells in spleens 48 hr after dosing. FIG.8C is a graph showing % of viable B cells in PBMC samples collected 6, 24 and 48 hr after dosing. FIG.8D is a graph showing the overall count of viable B cells in PBMC samples collected 6, 24 and 48 hrs after dosing. DETAILED DESCRIPTION I. Introduction

[0037] The instant specification describes compositions, methods, processes, kits and devices for the selection, design, preparation, manufacture, formulation, and / or use of LNP-based RNA medicines (e.g., vaccines, gene therapies, or gene-editing therapeutics). In various embodiments, the LNP-based RNA medicines comprise an LNP delivery system (as described in detail herein) and an encapsulated cargo / payload (e.g., RNA in the case of RNA medicines).

[0038] In various embodiments and as described further herein, the LNP delivery vehicle is a complex nanostructured body that provides protection to an encapsulated RNA payload (i.e., one or more RNA molecules) environmental damage (e.g., an intracellular environment). LNPs are formed through self-assembly of multiple lipid components, including (i) an ionizable lipid (e.g., ALC-0315 as in COMIRNATY® (Pfizer-BioNTech), SM-102 as in SPIKEVAX® (Moderna), or MC3 as in ONPATTRO® (Alnylam), or those ionizable lipids described herein), (ii) a helper lipid (such as, butnot limited to, 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC)), (iii) a sterol (e.g., cholesterol), and (iv) a PEG-lipid (e.g., PEG-DSPE).

[0039] In various embodiments and as described further herein, the RNA payload in the herein described LNP-based medicines may comprise coding and / or non-coding RNA, and / or mixtures thereof. The particular RNA payload constituents will generally reflect the medicine. For example, an LNP-based vaccine or therapeutic may comprise only coding RNA for expressing a vaccine antigen or a therapeutic protein, respectively. However, an LNP-based gene editing medicine may comprise a combination of coding RNA (e.g., encoding a CRISPR nuclease) and non-coding RNAs (e.g., guide RNAs). In various embodiments, the RNA molecule delivered by the herein disclosed LNPs can be a linear mRNA. In other embodiments, the RNA molecule delivered by the herein disclosed LNPs can be a circular mRNA. In still other embodiments, the RNA molecule delivered by the herein disclosed LNPs can include both linear and circular forms of mRNA. In further embodiments, the RNA may comprise one or more modifications, including chemical modifications (e.g., ribonucleotide analogs, alternative phosphate chain linkers), sequence modification (e.g., relative to a wild type sequence), and / or structural modification (e.g., secondary- folded structures, such as, but not limited to, stem-loops, hairpins, and G-quadruplexes, and tertiary structural elements, such as, but not limited to, helical duplexes and triple-stranded structures). A. LNP-Based RNA Vaccines

[0040] Described herein in certain aspects are improved LNP-based RNA vaccines for use in immunization against disease. In various aspects, the disclosure describes improved LNPs, including better performing ionizable lipids, that enhance the targeted delivery of LNP-based RNA vaccines and therapeutics based on linear and / or circular mRNAs. The improved LNPs protect linear and / or circular mRNA cargos (i.e., the circular and / or linear mRNA molecules encapsulated by the LNPs) from degradation and clearance while achieving targeted systemic or local delivery for use as enhanced vaccines.

[0041] The instant specification describes compositions, methods, processes, kits and devices for the selection, design, preparation, manufacture, formulation, and / or use of LNP-based RNA vaccines. In particular, as described herein are compositions, methods, processes, kits and devices for the selection, design, preparation, manufacture, formulation, and / or use of LNP-based RNA vaccines for the delivery of an RNA molecule that codes for one or more immunogenic viral antigens for use as vaccine and / or immunogenic compositions. In various embodiments, the RNA molecules delivered by the herein disclosed LNPs can be linear mRNA. In other embodiments, the RNA molecules delivered by the herein disclosed LNPs can be circular mRNA. In still other embodiments, the RNA molecule delivered by the herein disclosed LNPs can include both linear and circular forms of mRNA. In further embodiments, the RNA may comprise one or more modifications, including chemical modifications (e.g., ribonucleotide analogs, alternative phosphate chain linkers), sequence modification (e.g., relative to a wild type sequence), and / or structural modification (e.g.,secondary-folded structures, such as, but not limited to, stem-loops, hairpins, and G-quadruplexes, and tertiary structural elements, such as, but not limited to, helical duplexes and triple-stranded structures). In various other embodiments, the disclosure provides novel lipid components of the herein disclosed LNPs, including, but not limited to, novel ionizable lipids. B. LNP-Based RNA Therapeutics

[0042] Described herein in certain aspects are improved LNP-based RNA therapeutics for use in treating disease or a symptom thereof. In various aspects, the disclosure describes improved LNPs, including better performing ionizable lipids, that enhance the targeted delivery of LNP-based RNA therapeutics based on linear and / or circular mRNAs. The improved LNPs protect linear and / or circular mRNA cargos (i.e., the circular and / or linear mRNA molecules encapsulated by the LNPs) from degradation and clearance while achieving targeted systemic or local delivery for use as enhanced therapeutic agents.

[0043] The instant specification describes compositions, methods, processes, kits and devices for the selection, design, preparation, manufacture, formulation, and / or use of LNP-based RNA therapeutics. In particular, described herein are compositions, methods, processes, kits and devices for the selection, design, preparation, manufacture, formulation, and / or use of LNP-based RNA therapeutics for the delivery of an RNA molecule that codes for one or more therapeutic proteins for use treating a disease or a symptom thereof. Further described herein are compositions, methods, processes, kits and devices for the selection, design, preparation, manufacture, formulation, and / or use of LNP-based RNA therapeutics for the administration of an RNA molecule that codes for one or more therapeutic proteins for the prophylactic and / or therapeutic treatment of one or more diseases or a symptom thereof. In various embodiments, the RNA molecules delivered by the herein disclosed LNPs can be linear mRNA. In other embodiments, the RNA molecules delivered by the herein disclosed LNPs can be circular mRNA. In still other embodiments, the RNA molecules delivered by the herein disclosed LNPs can include both linear and circular forms of mRNA. In further embodiments, the RNA may comprise one or more modifications, including chemical modifications (e.g., ribonucleotide analogs, alternative phosphate chain linkers), sequence modification (e.g., relative to a wild type sequence), and / or structural modification (e.g., secondary- folded structures, such as, but not limited to, stem-loops, hairpins, and G-quadruplexes, and tertiary structural elements, such as, but not limited to, helical duplexes and triple-stranded structures). In various other embodiments, the disclosure provides novel lipid components of the herein disclosed LNPs, including, but not limited to, novel ionizable lipids. C. LNP-Based Gene Editing Therapeutics

[0044] Also described herein are LNP compositions comprising gene editing systems for use in treating disease and / or otherwise modifying the sequence and / or expression of target nucleotide sequences. The disclosure provides LNPs capable of delivering a gene editing system to a target organ, tissue, and / or cell. The gene editing systems may be delivered to cells under in vitro or ex vivoconditions and to organs, tissues, or cells under in vivo conditions (e.g., administered to a subject in an effective amount).

[0045] The disclosure also provides in various aspects therapeutic or pharmaceutical compositions comprising LNPs comprising gene editing systems or one or more components thereof. The gene editing systems may comprise DNA components, RNA components, protein components, nucleoprotein components, polysaccharide components, or combinations thereof. In other aspects, the disclosure provides nucleic acid molecules (e.g., RNA or DNA) that encode and / or constitute various componentry of the deliverable gene editing systems contemplated herein. In addition, other aspects of the disclosure provide nucleic acid molecules as components of the herein contemplated gene editing systems, such as, but not limited to plasmids or vectors encoding one or more components of a gene editing system, RNAs encoding one or more components of a gene editing system (e.g., mRNAs coding for a nuclease domain of a gene editing system), and non-coding RNAs (e.g., guide RNAs capable of complexing with and targeting a nucleic acid-programmable DNA binding domain to a specific target nucleotide sequence or a retron ncRNAs).

[0046] In further embodiments, the nucleic acid components (e.g., RNA) may comprise one or more modifications, including chemical modifications (e.g., ribonucleotide analogs, alternative phosphate chain linkers), sequence modification (e.g., relative to a wild type sequence), and / or structural modification (e.g., secondary-folded structures, such as, but not limited to, stem-loops, hairpins, and G-quadruplexes, and tertiary structural elements, such as, but not limited to, helical duplexes and triple-stranded structures).

[0047] The disclosure, in other aspects, describes various protein components (which may be encoded by the nucleic acid components described herein) of the various gene editing systems contemplated herein, including, but not limited to, user-programmable DNA binding proteins and various effector proteins, such as nucleases, polymerases, reverse transcriptases, recombinases, integrases, endonucleases, exonucleases, transposases, and deaminases.

[0048] The disclosure also describes nucleoprotein components of the gene editing systems contemplated herein, such as, but not limited to nuclease-guide RNA complexes. The disclosure also provides methods of modifying the sequence and / or expression level of a target nucleic acid molecule through the delivery and / or administration of an LNP described herein that comprises a gene editing system or components thereof. Still further, the disclosure provides methods of treating a disease by administering a therapeutically effective amount of an LNP-based gene editing system that results in the modification in the sequence and / or expression level of a target nucleic acid molecule (e.g., a disease-associated gene or regulatory sequence, such as a promoter, transcription factor binding site, or gene enhancer site).

[0049] The gene editing systems deliverable by the herein disclosed LNPs can be any type of gene editing system. Without limitation, the gene editing systems contemplated herein can include (A) nucleobase gene editing systems which result in one or more the changes to the sequence of atarget nucleic acid molecule (e.g., a gene or gene regulatory sequence) (sequence modifications may include, but are not limited to, an insertion of one or more base pairs, a deletion of one of more base pairs, a substitution or one or more base pairs, a conversion of a base pair to another base pair (e.g., a G:C pair converted to an A:T pair), an inversion, or a translocation), (B) an epigenetic editing system which results in one or more modifications to the epigenome to bring about an effect on gene expression without altering the sequence of a nucleic acid molecule, and (C) gene editing systems that combine the features of nucleobase editing systems and epigenetic editing systems (e.g., combining components from both types of systems to change the sequence and an epigenomic component with one system).

[0050] Nucleobase editing systems include a wide array of configurations with various combinations of protein functionalities and / or nucleic acid molecule components, all of which are contemplated herein. In general, nucleobase editing systems comprise at least a (i) DNA binding domain that is user-programmable to target a specific sequence in a nucleic acid molecule and optionally (ii) one or more effector domains that facilitate the modification of the sequence of the nucleic acid molecule. User-programmability may comprise amino acid sequence-programmable DNA binding domains (e.g., TALENS, zinc finger-binding domains, meganucleases (or homing endonucleases)) or nucleic acid sequence-programmable DNA binding domains or proteins (“naspDBP”) (e.g., CRISPR-Cas9, CRISPR-Cas12a, CRISPR-Cas12f, CRISPR-Cas13a, CRISPR- Cas13b, or TnpB).

[0051] Similarly, epigenetic editing systems comprise at least a (i) DNA binding domain that targets a specific sequence in a nucleic acid molecule and (ii) one or more effector domains that facilitates the modification of one or more epigenomic features of the nucleic acid molecule.

[0052] Gene editing systems may comprise one or more effector domains that provide various functionalities that facilitate changes in nucleotide sequence and / or gene expression, such as, but not limited to, single-strand DNA binding proteins, nucleases, endonucleases, exonucleases, deaminases (e.g., cytidine deaminases or adenosine deaminases), polymerases (e.g., reverse transcriptases), integrases, recombinases, etc., and fusion proteins comprising one or more functional domains linked together.

[0053] In addition, gene editing systems that utilize a nucleic acid sequence-programmable DNA binding domain or protein (naspDBP) may also comprise one or more non-coding nucleic acids, such as, one or more guide RNAs which complex with the nucleic acid programmable DNA binding protein (naspDBP) and target the complex to a specific nucleotide sequence. In the case of prime editing, the guide RNA may be a prime editing guide RNA (“pegRNA”) which comprises a specialized RNA template molecule that provides a template or coding sequence for a reverse transcriptase of the prime editing system. In some embodiments, the RNA template molecule may be coupled to a guide RNA as an extension arm at the 5’ or 3’ end of the guide RNA. In other embodiments, the RNA template molecule may be provided in trans as a separate molecule in amanner such that the RNA template molecule may itself become localized and associated with the target sequence and / or the gene editing system at the site of editing. In some embodiments, co- localization of an in trans RNA template molecule may be achieved with an aptamer or other RNA structure which binds to a binding partner that is coupled to, integrated with, or otherwise associated with the editing complex.

[0054] In the case of editing systems comprising a nucleic acid sequence-programmable DNA binding protein (naspDBP), such as a CRISPR-Cas9 or CRISPR-Cas12a nuclease, appropriate guides may be designed and synthesized using methods, software, and commercial sources which are well known to those having ordinary skill in the art such that guide RNAs for any given naspDBP may be obtained without undue experimentation.

[0055] Reference may be made to the following references providing information and tools for the design, synthesis, modification, and structural configuration of guide RNAs: (1) Mohr SE, Hu Y, Ewen-Campen B, Housden BE, Viswanatha R, Perrimon N. CRISPR guide RNA design for research applications. FEBS J.2016 Sep;283(17):3232-8. doi: 10.1111 / febs.13777. Epub 2016 Jun 22. PMID: 27276584; PMCID: PMC5014588; (2) Hoberecht L, Perampalam P, Lun A, Fortin JP. A comprehensive Bioconductor ecosystem for the design of CRISPR guide RNAs across nucleases and technologies. Nat Commun.2022 Nov 2;13(1):6568. doi: 10.1038 / s41467-022-34320-7. PMID: 36323688; PMCID: PMC9630310; (3) Cram D, Kulkarni M, Buchwaldt M, Rajagopalan N, Bhowmik P, Rozwadowski K, Parkin IAP, Sharpe AG, Kagale S. WheatCRISPR: a web-based guide RNA design tool for CRISPR / Cas9-mediated genome editing in wheat. BMC Plant Biol.2019 Nov 6;19(1):474. doi: 10.1186 / s12870-019-2097-z. PMID: 31694550; PMCID: PMC6836449; (4) Pliatsika V, Rigoutsos I. "Off-Spotter": very fast and exhaustive enumeration of genomic lookalikes for designing CRISPR / Cas guide RNAs. Biol Direct.2015 Jan 29;10:4. doi: 10.1186 / s13062-015- 0035-z. PMID: 25630343; PMCID: PMC4326336; (5) Hoof JB, Nødvig CS, Mortensen UH. Genome Editing: CRISPR-Cas9. Methods Mol Biol.2018;1775:119-132. doi: 10.1007 / 978-1-4939-7804-5_11. PMID: 29876814; (6) Labun K, Krause M, Torres Cleuren Y, Valen E. CRISPR Genome Editing Made Easy Through the CHOPCHOP Website. Curr Protoc.2021 Apr;1(4):e46. doi: 10.1002 / cpz1.46. PMID: 33905612; (7) Lee CM, Davis TH, Bao G. Examination of CRISPR / Cas9 design tools and the effect of target site accessibility on Cas9 activity. Exp Physiol.2018 Apr 1;103(4):456-460. doi: 10.1113 / EP086043. Epub 2017 Apr 12. PMID: 28303677; PMCID: PMC7266697; (8) Ma S, Lv J, Feng Z, Rong Z, Lin Y. Get ready for the CRISPR / Cas system: A beginner's guide to the engineering and design of guide RNAs. J Gene Med.2021 Nov;23(11):e3377. doi: 10.1002 / jgm.3377. Epub 2021 Jul 28. PMID: 34270141; (9) Hiranniramol K, Chen Y, Wang X. CRISPR / Cas9 Guide RNA Design Rules for Predicting Activity. Methods Mol Biol.2020;2115:351- 364. doi: 10.1007 / 978-1-0716-0290-4_19. PMID: 32006410; (10) Wiles MV, Qin W, Cheng AW, Wang H. CRISPR-Cas9-mediated genome editing and guide RNA design. Mamm Genome.2015 Oct;26(9-10):501-10. doi: 10.1007 / s00335-015-9565-z. Epub 2015 May 20. PMID: 25991564;PMCID: PMC4602062; (11) Creutzburg SCA, Wu WY, Mohanraju P, Swartjes T, Alkan F, Gorodkin J, Staals RHJ, van der Oost J. Good guide, bad guide: spacer sequence-dependent cleavage efficiency of Cas12a. Nucleic Acids Res.2020 Apr 6;48(6):3228-3243. doi: 10.1093 / nar / gkz1240. PMID: 31989168; PMCID: PMC7102956; (12) Heigwer F, Boutros M. Cloud-Based Design of Short Guide RNA (sgRNA) Libraries for CRISPR Experiments. Methods Mol Biol.2021;2162:3-22. doi: 10.1007 / 978-1-0716-0687-2_1. PMID: 32926374; (13) Dronina J, Samukaite-Bubniene U, Ramanavicius A. Towards application of CRISPR-Cas12a in the design of modern viral DNA detection tools (Review). J Nanobiotechnology.2022 Jan 21;20(1):41. doi: 10.1186 / s12951-022- 01246-7. PMID: 35062978; PMCID: PMC8777428; (14) Krysler AR, Cromwell CR, Tu T, Jovel J, Hubbard BP. Guide RNAs containing universal bases enable Cas9 / Cas12a recognition of polymorphic sequences. Nat Commun.2022 Mar 25;13(1):1617. doi: 10.1038 / s41467-022-29202-x. PMID: 35338140; PMCID: PMC8956631; (15) Shin HR, Kweon J, Kim Y. Gene Manipulation Using Fusion Guide RNAs for Cas9 and Cas12a. Methods Mol Biol.2021;2162:185-193. doi: 10.1007 / 978- 1-0716-0687-2_10. PMID: 32926383; (16) Schubert MS, Thommandru B, Woodley J, Turk R, Yan S, Kurgan G, McNeill MS, Rettig GR. Optimized design parameters for CRISPR Cas9 and Cas12a homology-directed repair. Sci Rep.2021 Sep 30;11(1):19482. doi: 10.1038 / s41598-021-98965-y. PMID: 34593942; PMCID: PMC8484621; (17) Crone MA, MacDonald JT, Freemont PS, Siciliano V. gDesigner: computational design of synthetic gRNAs for Cas12a-based transcriptional repression in mammalian cells. NPJ Syst Biol Appl.2022 Sep 16;8(1):34. doi: 10.1038 / s41540-022-00241-w. PMID: 36114193; PMCID: PMC9481559; (18) Konstantakos V, Nentidis A, Krithara A, Paliouras G. CRISPR-Cas9 gRNA efficiency prediction: an overview of predictive tools and the role of deep learning. Nucleic Acids Res.2022 Apr 22;50(7):3616-3637. doi: 10.1093 / nar / gkac192. PMID: 35349718; PMCID: PMC9023298; (19) Wang J, Zhang X, Cheng L, Luo Y. An overview and metanalysis of machine and deep learning-based CRISPR gRNA design tools. RNA Biol.2020 Jan;17(1):13-22. doi: 10.1080 / 15476286.2019.1669406. Epub 2019 Sep 27. PMID: 31533522; PMCID: PMC6948960; and (20) Cram D, Kulkarni M, Buchwaldt M, Rajagopalan N, Bhowmik P, Rozwadowski K, Parkin IAP, Sharpe AG, Kagale S. WheatCRISPR: a web-based guide RNA design tool for CRISPR / Cas9-mediated genome editing in wheat. BMC Plant Biol.2019 Nov 6;19(1):474. doi: 10.1186 / s12870-019-2097-z. PMID: 31694550; PMCID: PMC6836449; each of which are incorporated herein by reference in their entireties.

[0056] In the case of prime editing, in particular, further reference may be made to the following references providing information and tools for the design, synthesis, modification, and structural configuration of pegRNAs: (1) Hsu JY, Grünewald J, Szalay R, Shih J, Anzalone AV, Lam KC, Shen MW, Petri K, Liu DR, Joung JK, Pinello L. PrimeDesign software for rapid and simplified design of prime editing guide RNAs. Nat Commun.2021 Feb 15;12(1):1034. doi: 10.1038 / s41467- 021-21337-7. PMID: 33589617; PMCID: PMC7884779; (2) Li Y, Chen J, Tsai SQ, Cheng Y. Easy- Prime: a machine learning-based prime editor design tool. Genome Biol.2021 Aug 19;22(1):235. doi:10.1186 / s13059-021-02458-0. PMID: 34412673; PMCID: PMC8377858; (3) Zhang W, Petri K, Ma J, Lee H, Tsai CL, Joung JK, Yeh JJ. Enhancing CRISPR prime editing by reducing misfolded pegRNA interactions. bioRxiv [Preprint].2023 Aug 15:2023.08.14.553324. doi: 10.1101 / 2023.08.14.553324. PMID: 37645936; PMCID: PMC10462064; (4) Jin S, Lin Q, Gao Q, Gao C. Optimized prime editing in monocot plants using PlantPegDesigner and engineered plant prime editors (ePPEs). Nat Protoc.2023 Mar;18(3):831-853. doi: 10.1038 / s41596-022-00773-9. Epub 2022 Nov 25. PMID: 36434096; (5) Lin Q, Jin S, Zong Y, Yu H, Zhu Z, Liu G, Kou L, Wang Y, Qiu JL, Li J, Gao C. High-efficiency prime editing with optimized, paired pegRNAs in plants. Nat Biotechnol.2021 Aug;39(8):923-927. doi: 10.1038 / s41587-021-00868-w. Epub 2021 Mar 25. PMID: 33767395; (6) Standage-Beier K, Tekel SJ, Brafman DA, Wang X. Prime Editing Guide RNA Design Automation Using PINE-CONE. ACS Synth Biol.2021 Feb 19;10(2):422-427. doi: 10.1021 / acssynbio.0c00445. Epub 2021 Jan 19. PMID: 33464043; PMCID: PMC7901017; (7) Zhang W, Petri K, Ma J, Lee H, Tsai CL, Joung JK, Yeh JJ. Enhancing CRISPR prime editing by reducing misfolded pegRNA interactions. bioRxiv [Preprint].2023 Aug 15:2023.08.14.553324. doi: 10.1101 / 2023.08.14.553324. PMID: 37645936; PMCID: PMC10462064; (8) Chow RD, Chen JS, Shen J, Chen S. A web tool for the design of prime-editing guide RNAs. Nat Biomed Eng.2021 Feb;5(2):190-194. doi: 10.1038 / s41551-020-00622-8. Epub 2020 Sep 28. PMID: 32989284; PMCID: PMC7882013; each of which are incorporated herein by reference in their entireties.

[0057] Reference may also be made to the following commercial vendors which sell guide RNAs for CRISPR editing applications (including base editing and prime editing) and provide various tools and instruction for the ordering, design, synthesis, modification, and structural configuration of guide RNAs: GENSCRIPT, SYNTHEGO, TAKARA BIO, INTEGRATED DNA TECHNOLOGIES, LC SCIENCES, HORIZON DISCOVERY; SIGMA-ALDRICH; ORIGENE, and TWIST BIOSCIENCES, among others.

[0058] In addition, guide RNA may be modified with chemical modifications and / or structural modifications for enhancing various properties thereof, including specificity, stability, and limiting off-target activity. One of ordinary skill in the art will be able to modify a guide RNA with any known modification without undue experimentation. Guide modifications are discussed in the following references: (1) Ke Y, Ghalandari B, Huang S, Li S, Huang C, Zhi X, Cui D, Ding X.2'-O- Methyl modified guide RNA promotes the single nucleotide polymorphism (SNP) discrimination ability of CRISPR-Cas12a systems. Chem Sci.2022 Feb 1;13(7):2050-2061. doi: 10.1039 / d1sc06832f. PMID: 35308857; PMCID: PMC8848812; (2) Allen D, Rosenberg M, Hendel A. Using Synthetically Engineered Guide RNAs to Enhance CRISPR Genome Editing Systems in Mammalian Cells. Front Genome Ed.2021 Jan 28;2:617910. doi: 10.3389 / fgeed.2020.617910. PMID: 34713240; PMCID: PMC8525374; (3) Basila M, Kelley ML, Smith AVB. Minimal 2'-O-methyl phosphorothioate linkage modification pattern of synthetic guide RNAs for increased stability and efficient CRISPR-Cas9 gene editing avoiding cellular toxicity. PLoS One.2017 Nov27;12(11):e0188593. doi: 10.1371 / journal.pone.0188593. PMID: 29176845; PMCID: PMC5703482; (4) Sakovina L, Vokhtantsev I, Vorobyeva M, Vorobyev P, Novopashina D. Improving Stability and Specificity of CRISPR / Cas9 System by Selective Modification of Guide RNAs with 2'-fluoro and Locked Nucleic Acid Nucleotides. Int J Mol Sci.2022 Nov 3;23(21):13460. doi: 10.3390 / ijms232113460. PMID: 36362256; PMCID: PMC9655745; (5) Shapiro J, Tovin A, Iancu O, Allen D, Hendel A. Chemical Modification of Guide RNAs for Improved CRISPR Activity in CD34+ Human Hematopoietic Stem and Progenitor Cells. Methods Mol Biol.2021;2162:37-48. doi: 10.1007 / 978-1-0716-0687-2_3. PMID: 32926376; (6) Filippova J, Matveeva A, Zhuravlev E, Stepanov G. Guide RNA modification as a way to improve CRISPR / Cas9-based genome-editing systems. Biochimie.2019 Dec;167:49-60. doi: 10.1016 / j.biochi.2019.09.003. Epub 2019 Sep 4. PMID: 31493470; (7) Hendel A, Bak RO, Clark JT, Kennedy AB, Ryan DE, Roy S, Steinfeld I, Lunstad BD, Kaiser RJ, Wilkens AB, Bacchetta R, Tsalenko A, Dellinger D, Bruhn L, Porteus MH. Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells. Nat Biotechnol.2015 Sep;33(9):985-989. doi: 10.1038 / nbt.3290. Epub 2015 Jun 29. PMID: 26121415; PMCID: PMC4729442; (8)_ Ryan DE, Taussig D, Steinfeld I, Phadnis SM, Lunstad BD, Singh M, Vuong X, Okochi KD, McCaffrey R, Olesiak M, Roy S, Yung CW, Curry B, Sampson JR, Bruhn L, Dellinger DJ. Improving CRISPR-Cas specificity with chemical modifications in single-guide RNAs. Nucleic Acids Res.2018 Jan 25;46(2):792-803. doi: 10.1093 / nar / gkx1199. Erratum in: Nucleic Acids Res.2022 Mar 21;50(5):2986. PMID: 29216382; PMCID: PMC5778453; (9) Palumbo CM, Gutierrez-Bujari JM, O'Geen H, Segal DJ, Beal PA. Versatile 3' Functionalization of CRISPR Single Guide RNA. Chembiochem.2020 Jun 2;21(11):1633-1640. doi: 10.1002 / cbic.201900736. Epub 2020 Mar 5. PMID: 31943634; PMCID: PMC7323579; (10) Mullally G, van Aelst K, Naqvi MM, Diffin FM, Karvelis T, Gasiunas G, Siksnys V, Szczelkun MD.5' modifications to CRISPR-Cas9 gRNA can change the dynamics and size of R-loops and inhibit DNA cleavage. Nucleic Acids Res.2020 Jul 9;48(12):6811-6823. doi: 10.1093 / nar / gkaa477. PMID: 32496535; PMCID: PMC7337959; (12) Lu S, Zhang Y, Yin H. Chimeric DNA-RNA Guide RNA Designs. Methods Mol Biol.2021;2162:79-85. doi: 10.1007 / 978-1-0716-0687-2_6. PMID: 32926379; each of which are incorporated by reference herein in their entireties.

[0059] In the specific case of prime editing, pegRNAs may be modified with chemical modifications and / or structural modifications for enhancing various properties thereof, including specificity, stability, and limiting off-target activity. One of ordinary skill in the art will be able to modify a pegRNA for prime editing with any known modification without undue experimentation. pegRNA modifications are discussed in the following references: (1) Nelson JW, Randolph PB, Shen SP, Everette KA, Chen PJ, Anzalone AV, An M, Newby GA, Chen JC, Hsu A, Liu DR. Engineered pegRNAs improve prime editing efficiency. Nat Biotechnol.2022 Mar;40(3):402-410. doi: 10.1038 / s41587-021-01039-7. Epub 2021 Oct 4. Erratum in: Nat Biotechnol.2021 Dec 8;: PMID: 34608327; PMCID: PMC8930418; (2) Liu B, Dong X, Cheng H, Zheng C, Chen Z, Rodríguez TC,Liang SQ, Xue W, Sontheimer EJ. A split prime editor with untethered reverse transcriptase and circular RNA template. Nat Biotechnol.2022 Sep;40(9):1388-1393. doi: 10.1038 / s41587-022-01255- 9. Epub 2022 Apr 4. PMID: 35379962; each of which are incorporated by reference herein in their entireties.

[0060] Other specialized guide RNAs may be included depending upon the requirements and / or nature of the gene editing system and the cognate nucleic acid programmable proteins. For example, TnpB enzymes require a specialized guide RNA referred to as reRNA. Also, guide RNAs have different characteristics (e.g., PAM preferences, the spacer length, and the scaffold portion that binds to the nuclease protein) depending upon the programmable nuclease requirements.

[0061] The gene editing systems contemplated here may introduce a wide variety of changes, including (A) a change in the sequence of the target nucleic acid molecule, such as, but not limited to, (i) a nucleobase substitution (e.g., a purine to a pyrimidine), (ii) a deletion of one or more nucleobases, (iii) an insertion of one or more nucleobases, (iv) a combination of a deletion and insertion of one or more nucleobases, (v) an inversion of a nucleobase sequence, a (vi) translocation of a nucleobase sequence, and (vii) a combination or two or more such modifications, and (B) one or more modifications to the epigenome to bring about an effect on gene expression without altering the sequence of a nucleic acid molecule wherein said epigenetic change results in altered gene expression through altered chromatin structure or accessibility.

[0062] The LNP compositions and / or gene editing systems described herein may include a variety of coding RNA molecules that code for the various components of gene editors. In various aspects, the coding RNA may be linear mRNA. In other embodiments, the coding RNA may be circular mRNA. In various aspects, the improved LNPs protect linear and / or circular mRNA cargos from degradation and clearance while achieving targeted systemic or local delivery for use as enhanced gene editing platforms and / or therapeutic agents.

[0063] In various other aspects, the LNP compositions and / or gene editing systems described herein may also include a repair template, e.g., an homology-directed repair (HDR)-dependent repair template (or HDR template). Such HDR templates are well-known in the art and can include single- strand or double-stranded DNA (e.g., oligos) or RNA. Further information regarding HDR and HDR templates for use in editing systems for various applications, such as gene knock-in, may be found in Fu YW, Dai XY, Wang WT, Yang ZX, Zhao JJ, Zhang JP, Wen W, Zhang F, Oberg KC, Zhang L, Cheng T, Zhang XB. Dynamics and competition of CRISPR-Cas9 ribonucleoproteins and AAV donor-mediated NHEJ, MMEJ and HDR editing. Nucleic Acids Res.2021 Jan 25;49(2):969-985. doi: 10.1093 / nar / gkaa1251. PMID: 33398341; PMCID: PMC7826255; Iyer S, Mir A, Vega-Badillo J, Roscoe BP, Ibraheim R, Zhu LJ, Lee J, Liu P, Luk K, Mintzer E, Guo D, Soares de Brito J, Emerson CP Jr, Zamore PD, Sontheimer EJ, Wolfe SA. Efficient Homology-Directed Repair with Circular Single-Stranded DNA Donors. CRISPR J.2022 Oct;5(5):685-701. doi: 10.1089 / crispr.2022.0058. Epub 2022 Sep 7. PMID: 36070530; PMCID: PMC9595650; and Richardson CD, Ray GJ, DeWittMA, Curie GL, Corn JE. Enhancing homology-directed genome editing by catalytically active and inactive CRISPR-Cas9 using asymmetric donor DNA. Nat Biotechnol.2016 Mar;34(3):339-44. doi: 10.1038 / nbt.3481. Epub 2016 Jan 20. PMID: 26789497, each of which are incorporated herein by reference in their entireties.

[0064] Accordingly, the instant specification describes compositions, methods, processes, kits and devices for the selection, design, preparation, manufacture, formulation, and / or use of LNP- based gene editing systems as therapeutic compositions. Further described herein are compositions, methods, processes, kits and devices for the selection, design, preparation, manufacture, formulation, and / or use of LNP-based gene editing therapeutics for the prophylactic and / or therapeutic treatment of one or more diseases or a symptom thereof. The components capable of being encapsulated by or otherwise incorporated by the LNPs described herein may be referred to as LNP “payloads” and may include all of the biological materials described above, including DNA molecules, RNA molecules (coding and / or non-coding), proteins, and nucleoproteins (e.g., Cas / guide RNA complexes). II. LNP delivery systems

[0065] The RNA payloads (e.g., linear and circular mRNAs) described herein may be encapsulated and delivered by lipid nanoparticles (LNPs) and compositions and / or formulations comprising RNA-encapsulated LNPs.

[0066] Below describes LNPs that may be used as the RNA payload delivery vehicles contemplated herein, as well as the various ionizable lipids, structural lipids, PEGylated lipids, and phospholipids that may be used to make the herein LNPs for delivery RNA payloads to cells. In addition, below describes additional LNP components that are contemplated, such as targeting moieties and other lipid components. A. Lipid Nanoparticle Compositions

[0067] In one aspect, the present disclosure further provides delivery systems for delivery of a therapeutic payload (e.g., the RNA payloads described herein which may encode a polypeptid of interest, e.g., an antigen or a therapeutic protein) disclosed herein. In some embodiments, a delivery system suitable for delivery of the therapeutic payload disclosed herein comprises a lipid nanoparticle (LNP) formulation.

[0068] In some embodiments, an LNP of the present disclosure comprises an ionizable lipid, a structural lipid, a PEGylated lipid (aka PEG lipid), and a phospholipid. In alternative embodiments, an LNP comprises an ionizable lipid, a structural lipid, a PEGylated lipid (aka PEG lipid), and a zwitterionic amino acid lipid. In some embodiments, an LNP further comprises a 5th lipid, besides any of the aforementioned lipid components. In some embodiments, the LNP encapsulates one or more elements of the active agent of the present disclosure. In some embodiments, an LNP further comprises a targeting moiety covalently or non-covalently bound to the outer surface of the LNP. In some embodiments, the targeting moiety is a targeting moiety that binds to, or otherwise facilitates uptake by, cells of a particular organ system.

[0069] In some embodiments, an LNP has a diameter of at least about 20nm, 30 nm, 40nm, 50nm, 60nm, 70nm, 80nm, or 90nm. In some embodiments, an LNP has a diameter of less than about 100nm, 110nm, 120nm, 130nm, 140nm, 150nm, or 160nm. In some embodiments, an LNP has a diameter of less than about 120 nm. In some embodiments, an LNP has a diameter of less than about 100nm. In some embodiments, an LNP has a diameter of less than about 90nm. In some embodiments, an LNP has a diameter of less than about 80nm. In some embodiments, an LNP has a diameter of about 60-100nm. In some embodiments, an LNP has a diameter of about 50-120nm. In some embodiments, an LNP has a diameter of about 75-80nm.

[0070] In some embodiments, the lipid nanoparticle compositions of the present disclosure are described according to the respective molar ratios of the component lipids in the formulation. As a non-limiting example, the mol-% of the ionizable lipid may be from about 10 mol-% to about 80 mol- %. As a non-limiting example, the mol-% of the ionizable lipid may be from about 20 mol-% to about 70 mol-%. As a non-limiting example, the mol-% of the ionizable lipid may be from about 30 mol-% to about 60 mol-%. As a non-limiting example, the mol-% of the ionizable lipid may be from about 35 mol-% to about 55 mol-%. As a non-limiting example, the mol-% of the ionizable lipid may be from about 40 mol-% to about 50 mol-%. As a non-limiting example, the mol-% of the ionizable lipid may be from about 30 mol-% to about 40 mol-%. As a non-limiting example, the mol-% of the ionizable lipid may be from about 25 mol-% to about 35 mol-%. In some embodiments, the mol-% of the ionizable lipid is about 10 mol-%. In some embodiments, the mol-% of the ionizable lipid is about 15 mol-%. In some embodiments, the mol-% of the ionizable lipid is about 20 mol-%. In some embodiments, the mol-% of the ionizable lipid is about 25 mol-%. In some embodiments, the mol-% of the ionizable lipid is about 30 mol-%. In some embodiments, the mol-% of the ionizable lipid is about 33 mol-%. In some embodiments, the mol-% of the ionizable lipid is about 35 mol-%. In some embodiments, the mol-% of the ionizable lipid is about 40 mol-%. In some embodiments, the mol-% of the ionizable lipid is about 45 mol-%. In some embodiments, the mol-% of the ionizable lipid is about 55 mol-%. In some embodiments, the mol-% of the ionizable lipid is about 60 mol-%.

[0071] In some embodiments, the mol-% of the phospholipid may be from about 1 mol-% to about 50 mol-%. In some embodiments, the mol-% of the phospholipid may be from about 2 mol-% to about 45 mol-%. In some embodiments, the mol-% of the phospholipid may be from about 3 mol- % to about 40 mol-%. In some embodiments, the mol-% of the phospholipid may be from about 4 mol-% to about 35 mol-%. In some embodiments, the mol-% of the phospholipid may be from about 5 mol-% to about 30 mol-%. In some embodiments, the mol-% of the phospholipid may be from about 10 mol-% to about 20 mol-%. In some embodiments, the mol-% of the phospholipid may be from about 5 mol-% to about 20 mol-%. In some embodiments, the mol-% of the phospholipid is from about 30 mol-% to about 60 mol-%. In some embodiments, the mol-% of the phospholipid is from about 35 mol-% to about 55 mol-%. In some embodiments, the mol-% of the phospholipid is from about 35 mol-% to about 45 mol-%. In some embodiments, the mol-% of the phospholipid is about 10mol-%. In some embodiments, the mol-% of the phospholipid is about 15 mol-%. In some embodiments, the mol-% of the phospholipid is about 20 mol-%. In some embodiments, the mol-% of the phospholipid is about 25 mol-%. In some embodiments, the mol-% of the phospholipid is about 30 mol-%. In some embodiments, the mol-% of the phospholipid is about 35 mol-%. In some embodiments, the mol-% of the phospholipid is about 40 mol-%. In some embodiments, the mol-% of the phospholipid is about 45 mol-%. In some embodiments, the mol-% of the phospholipid is about 55 mol-%. In some embodiments, the mol-% of the phospholipid is about 60 mol-%.

[0072] In some embodiments, the mol-% of the phospholipid as described above comprises two or more phospholipids at an individual mol-% that totals to an aforementioned amount. In certain embodiments, the mol-% of the phospholipid is about 20 mol-% each of two phospholipids. In certain embodiments, the mol-% of the phospholipid is about 15 mol-% each of two phospholipids. In certain embodiments, the mol-% of the phospholipid is about 25 mol-% each of two phospholipids. In certain embodiments, the mol-% of the phospholipid is about 30 mol-% each of two phospholipids. In certain embodiments, the mol-% of the phospholipid is about 15 mol-% of a first phospholipid and about 20 mol-% of a second phospholipid. In certain embodiments, the mol-% of the phospholipid is about 30 mol-% of a first phospholipid and about 10 mol-% of a second phospholipid. In certain embodiments, the mol-% of the phospholipid is about 25 mol-% of a first phospholipid and about 10 mol-% of a second phospholipid. In certain embodiments, the mol-% of the phospholipid is about 25 mol-% of a first phospholipid and about 20 mol-% of a second phospholipid. In certain embodiments, the mol-% of the phospholipid is about 15 mol-% of a first phospholipid and about 20 mol-% of a second phospholipid.

[0073] In some embodiments, the mol-% of the structural lipid may be from about 10 mol-% to about 80 mol-%. In some embodiments, the mol-% of the structural lipid may be from about 20 mol-% to about 70 mol-%. In some embodiments, the mol-% of the structural lipid may be from about 30 mol-% to about 60 mol-%. In some embodiments, the mol-% of the structural lipid may be from about 35 mol-% to about 55 mol-%. In some embodiments, the mol-% of the structural lipid may be from about 40 mol-% to about 50 mol-%.

[0074] In some embodiments, the mol-% of the PEG lipid may be from about 0.1 mol-% to about 10 mol-%. In some embodiments, the mol-% of the PEG lipid may be from about 0.2 mol-% to about 5 mol-%. In some embodiments, the mol-% of the PEG lipid may be from about 0.5 mol-% to about 3 mol-%. In some embodiments, the mol-% of the PEG lipid may be from about 1 mol-% to about 2 mol-%. In some embodiments, the mol-% of the PEG lipid may be about 1.5 mol-%. In some embodiments, the mol-% of the PEG lipid may be about 2.5 mol-%. i. Ionizable lipids

[0075] In some embodiments, an LNP disclosed herein comprises an ionizable lipid. In some embodiments, an LNP comprises two or more ionizable lipids.

[0076] Described below are a number of exemplary ionizable Lipids of the Disclosure.Formula (AX’’’’)

[0077] The present disclosure, in some embodiments, provides compounds of Formula (AX’’’’)or a pharmaceutically acceptable salt thereof, wherein: A is selected from an optionally substituted bridged carbocyclic bicycle, bridged carbocyclic multicycle, bridged heterocyclic bicycle, or bridged heterocyclic multicycle; n is an integer selected from 0, 1 or 2; m is an integer selected from 1 or 2, such that m plus n is less than or equal to 3; R1is selected from the group consisting of -OH, -OAc, -NR2, -N(R)RH,each R is independently -H or C1-C6aliphatic; each RHis C1-C6aliphatic-OH; each X1and XAare each independently a bond or optionally substituted C1-C6aliphatic; each Y1is independently selected from the group consisting of,, and a bond; wherein the bond marked with an "*" is attached to X1; each X2and X3is independently a bond or optionally substituted C1-C12 aliphatic; each Y2and Y3is independently selected from the group consisting ofwherein the bond marked with an "*" is attached to X2or X3; each X4and X5is independently a bond or optionally substituted C1-C6aliphatic; each Y4and Y5is independently selected from the group consisting of a bond,wherein the bond marked with an "*" is attached to X4or X5; each X6and X7is independently a bond or optionally substituted C1-C6 aliphatic; R2is -CH(OR6)(OR7), -CH(SR6)(SR7), -CH(R6)(R7), -CF(R6)(R7), -R10, optionally substituted C5-C18 aliphatic, or optionally substituted C1-C14 aliphatic-R10, wherein one or more methylene linkages of R2are each optionally and independently replaced with an optionally substituted C3-C8cycloalkylenyl, phenyl, -O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or -C(O)O-; each R3is independently -CH(OR8)(OR9), -CH(SR8)(SR9), -CH(R8)(R9), -CF(R8)(R9), -R11, optionally substituted C5-C18 aliphatic, or optionally substituted C1-C14 aliphatic-R11, wherein one or more methylene linkages of R3are each optionally and independently replaced with an optionally substituted C3-C8 cycloalkylenyl, phenyl, -O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, - NHC(O)-, or -C(O)O-; R6and R7are each independently optionally substituted -C1-C14aliphatic, -R10, or optionally substituted -C1-C14aliphatic-R10; wherein one or more methylene linkages of R6and R7are each optionally and independently replaced with an optionally substituted C3-C8cycloalkylenyl, phenyl, - O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or -C(O)O-; R8and R9are each independently optionally substituted -C1-C14 aliphatic, -R11, or optionally substituted -C1-C14 aliphatic-R11; wherein one or more methylene linkages of R8and R9are each optionally and independently replaced with an optionally substituted C3-C8cycloalkylenyl, phenyl, - O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or -C(O)O-; and each R10and R11is independently an optionally substituted cylic, bicyclic, bridged bicyclic, multicyclic or bridged multicyclic C4-C14cycloalkyl or optionally substituted cylic, bicyclic, bridged bicyclic, multicyclic or bridged multicyclic 4-14 membered heterocyclyl, or two R10or two R11taken together form an optionally substituted bridged bicyclic or multicyclic C4-C14 cycloalkyl or optionally substituted bridged bicyclic or multicyclic 4-14 membered heterocyclyl;wherein one or more of X1, XA, X2, X3, X4, X5, X6, X7, R2and R3is optionally and independently substituted with one or more substituents selected from -F, -Cl, -Br and -I. Formula (AX’’’)

[0078] The present disclosure, in some embodiments, provides compounds of Formula (AX’’’)or a pharmaceutically acceptable salt thereof, wherein: A is selected from an optionally substituted 4-14 membered bridged carbocyclic bicycle, bridged carbocyclic multicycle, bridged heterocyclic bicycle, or bridged heterocyclic multicycle; n is an integer selected from 0, 1 or 2; m is an integer selected from 1 or 2, such that m plus n is less than or equal to 3; R1is selected from the group consisting of -OH, -OAc, -NR H 2, -N(R)R ,each R is independently -H or C1-C6aliphatic; each RHis C1-C6aliphatic-OH; each X1and XAare each independently a bond or optionally substituted C1-C6 aliphatic; each Y1is independently selected from the group consisting of,, and a bond; wherein the bond marked with an "*" is attached to X1; each X2and X3is independently a bond or optionally substituted C1-C12 aliphatic; each Y2and Y3is independently selected from the group consisting ofwherein the bond marked with an "*" is attached to X2or X3; each X4and X5is independently a bond or optionally substituted C1-C6aliphatic; each Y4and Y5is independently selected from the group consisting of a bond,wherein the bond marked with an "*" is attached to X4or X5; each X6and X7is independently a bond or optionally substituted C1-C6 aliphatic; R2is -CH(OR6)(OR7), -CH(SR6)(SR7), -CH(R6)(R7), -CF(R6)(R7), -R10, optionally substituted C5-C18 aliphatic, or optionally substituted C1-C14 aliphatic-R10, wherein one or more methylene linkages of R2are each optionally and independently replaced with an optionally substituted C3-C8cycloalkylenyl, phenyl, -O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or -C(O)O-; each R3is independently -CH(OR8)(OR9), -CH(SR8)(SR9), -CH(R8)(R9), -CF(R8)(R9), -R11, optionally substituted C5-C18 aliphatic, or optionally substituted C1-C14 aliphatic-R11, wherein one or more methylene linkages of R3are each optionally and independently replaced with an optionally substituted C3-C8 cycloalkylenyl, phenyl, -O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, - NHC(O)-, or -C(O)O-; R6and R7are each independently optionally substituted -C1-C14aliphatic, -R10, or optionally substituted -C1-C14aliphatic-R10; wherein one or more methylene linkages of R6and R7are each optionally and independently replaced with an optionally substituted C3-C8cycloalkylenyl, phenyl, - O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or -C(O)O-; R8and R9are each independently optionally substituted -C1-C14 aliphatic, -R11, or optionally substituted -C1-C14 aliphatic-R11; wherein one or more methylene linkages of R8and R9are each optionally and independently replaced with an optionally substituted C3-C8cycloalkylenyl, phenyl, - O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or -C(O)O-; and each R10and R11is independently an optionally substituted cylic, bicyclic, bridged bicyclic, multicyclic or bridged multicyclic C4-C14cycloalkyl or optionally substituted cylic, bicyclic, bridged bicyclic, multicyclic or bridged multicyclic 4-14 membered heterocyclyl, or two R10or two R11taken together form an optionally substituted bridged bicyclic or multicyclic C4-C14 cycloalkyl or optionally substituted bridged bicyclic or multicyclic 4-14 membered heterocyclyl;wherein one or more of X1, XA, X2, X3, X4, X5, X6, X7, R2and R3is optionally and independently substituted with one or more substituents selected from -F, -Cl, -Br and -I. Formula (AX’’)

[0079] The present disclosure, in some embodiments, provides compounds of Formula (AX’’)or a pharmaceutically acceptable salt thereof, wherein: A is selected from an optionally substituted 4-14 membered bridged carbocyclic bicycle, bridged carbocyclic multicycle, bridged heterocyclic bicycle, or bridged heterocyclic multicycle; n is an integer selected from 0, 1 or 2; m is an integer selected from 1 or 2, such that m plus n is less than or equal to 3; R1is selected from the group consisting of -OH, -OAc, -NR2,each R is independently -H or C1-C6aliphatic; X1and XAare each independently a bond or optionally substituted C1-C6aliphatic; Y1is selected from the group consisting of,, and a bond; wherein the bond marked with an "*" is attached to X1; each X2and X3is independently a bond or optionally substituted C1-C12 aliphatic; each Y2and Y3is independently selected from the group consisting ofwherein the bond marked with an "*" is attached to X2or X3; each X4and X5is independently optionally substituted C1-C6aliphatic; each Y4and Y5is independently selected from the group consisting of a bond,wherein the bond marked with an "*" is attached to X4or X5; each X6and X7is independently a bond or optionally substituted C1-C6 aliphatic; R2is -CH(OR6)(OR7), -CH(SR6)(SR7), -CH(R6)(R7), -R10, optionally substituted C5-C18 aliphatic, or optionally substituted C1-C14 aliphatic-R10, wherein one or more methylene linkages of R2are each optionally and independently replaced with an optionally substituted C3-C8cycloalkylenyl, phenyl, - O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or -C(O)O-; each R3is independently -CH(OR8)(OR9), -CH(SR8)(SR9), -CH(R8)(R9), -R11, optionally substituted C5-C18 aliphatic, or optionally substituted C1-C14 aliphatic-R11, wherein one or more methylene linkages of R3are each optionally and independently replaced with an optionally substituted C3-C8 cycloalkylenyl, phenyl, -O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or -C(O)O-; R6and R7are each independently optionally substituted -C1-C14 aliphatic, -R10, or optionally substituted -C1-C14aliphatic-R10; wherein one or more methylene linkages of R6and R7are each optionally and independently replaced with an optionally substituted C3-C8cycloalkylenyl, phenyl, - O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or -C(O)O-; R8and R9are each independently optionally substituted -C1-C14 aliphatic, -R11, or optionally substituted -C1-C14 aliphatic-R11; wherein one or more methylene linkages of R8and R9are each optionally and independently replaced with an optionally substituted C3-C8 cycloalkylenyl, phenyl, - O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or -C(O)O-; and each R10and R11is independently an optionally substituted cylic, bicyclic, bridged bicyclic, multicyclic or bridged multicyclic C4-C14cycloalkyl or optionally substituted cylic, bicyclic, bridged bicyclic, multicyclic or bridged multicyclic 4-14 membered heterocyclyl, or two R10or two R11taken together form an optionally substituted bridged bicyclic or multicyclic C4-C14 cycloalkyl or optionally substituted bridged bicyclic or multicyclic 4-14 membered heterocyclyl.Formula (AX’)

[0080] The present disclosure, in some embodiments, provides compounds of Formula (AX’)(AX’), or a pharmaceutically acceptable salt thereof, wherein: A is selected from an optionally substituted 4-14 membered bridged carbocyclic bicycle, bridged carbocyclic multicycle, bridged heterocyclic bicycle, or bridged heterocyclic multicycle; n is an integer selected from 1 or 2; R1is selected from the group consisting of -OH, -OAc, -NR2,each R is independently -H or C1-C6 aliphatic; X1and XAare each independently a bond or optionally substituted C1-C6 aliphatic; Y1is selected from the group consisting of,, and a bond; wherein the bond marked with an "*" is attached to X1; each X2and X3is independently a bond or optionally substituted C1-C12aliphatic; each Y2and Y3is independently selected from the group consisting of; wherein the bond marked with an "*" is attached to X2or X3; each X4and X5is independently optionally substituted C1-C6aliphatic;R2is -CH(OR6)(OR7), -CH(SR6)(SR7), -CH(R6)(R7), -R10, optionally substituted C5-C18 aliphatic, or optionally substituted C1-C14 aliphatic-R10, wherein one or more methylene linkages of R2are each optionally and independently replaced with an optionally substituted C3-C8 cycloalkylenyl, phenyl, - O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or -C(O)O-; each R3is independently -CH(OR8)(OR9), -CH(SR8)(SR9), -CH(R8)(R9), -R11, optionally substituted C5-C18aliphatic, or optionally substituted C1-C14aliphatic-R11, wherein one or more methylene linkages of R3are each optionally and independently replaced with an optionally substituted C3-C8 cycloalkylenyl, phenyl, -O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or -C(O)O-; R6and R7are each independently optionally substituted -C1-C14 aliphatic, -R10, or optionally substituted -C1-C14 aliphatic-R10; wherein one or more methylene linkages of R6and R7are each optionally and independently replaced with an optionally substituted C3-C8cycloalkylenyl, phenyl, - O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or -C(O)O-; R8and R9are each independently optionally substituted -C1-C14aliphatic, -R11, or optionally substituted -C1-C14 aliphatic-R11; wherein one or more methylene linkages of R8and R9are each optionally and independently replaced with an optionally substituted C3-C8 cycloalkylenyl, phenyl, - O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or -C(O)O-; and each R10and R11is independently an optionally substituted cylic, bicyclic, bridged bicyclic, multicyclic or bridged multicyclic C4-C14cycloalkyl or optionally substituted cylic, bicyclic, bridged bicyclic, multicyclic or bridged multicyclic 4-14 membered heterocyclyl, or two R10or two R11taken together form an optionally substituted bridged bicyclic or multicyclic C4-C14cycloalkyl or optionally substituted bridged bicyclic or multicyclic 4-14 membered heterocyclyl. Formula (AX*)

[0081] The present disclosure, in some embodiments, provides compounds of Formula (AX*)(AX*), or a pharmaceutically acceptable salt thereof, wherein:A is selected from an optionally substituted bridged carbocyclic or heterocyclic core selected from then is an integer selected from 0, 1 or 2; m is an integer selected from 1 or 2, such that m plus n is less than or equal to 3; R1is selected from the group consisting of -OH, -OAc, -NR2, -N(R)RH ,each R is independently -H or C1-C6 aliphatic; each RHis C1-C6 aliphatic-OH; each X1and XAare each independently a bond or optionally substituted C1-C6aliphatic; each Y1is independently selected from the group consisting of,, and a bond; wherein the bond marked with an "*" is attached to X1; each X2and X3is independently a bond or optionally substituted C1-C12aliphatic; each Y2and Y3is independently selected from the group consisting of; wherein the bond marked with an "*" is attached to X2or X3; each X4and X5is independently a bond or optionally substituted C1-C6aliphatic; each Y4and Y5is independently selected from the group consisting of a bond,wherein the bond marked with an "*" is attached to X4or X5; each X6and X7is independently a bond or optionally substituted C1-C6aliphatic; R2is -CH(OR6)(OR7), -CH(SR6)(SR7), -CH(R6)(R7), -CF(R6)(R7), -R10, optionally substituted C5-C18 aliphatic, or optionally substituted C1-C14 aliphatic-R10, wherein one or more methylene linkages of R2are each optionally and independently replaced with an optionally substituted C3-C8 cycloalkylenyl, phenyl, -O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or -C(O)O-; each R3is independently -CH(OR8)(OR9), -CH(SR8)(SR9), -CH(R8)(R9), -CF(R8)(R9), -R11, optionally substituted C5-C18aliphatic, or optionally substituted C1-C14aliphatic-R11, wherein one or more methylene linkages of R3are each optionally and independently replaced with an optionally substituted C3-C8 cycloalkylenyl, phenyl, -O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, - NHC(O)-, or -C(O)O-; R6and R7are each independently optionally substituted -C1-C14 aliphatic, -R10, or optionally substituted -C1-C14aliphatic-R10; wherein one or more methylene linkages of R6and R7are each optionally and independently replaced with an optionally substituted C3-C8cycloalkylenyl, phenyl, - O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or -C(O)O-; R8and R9are each independently optionally substituted -C1-C14 aliphatic, -R11, or optionally substituted -C1-C14 aliphatic-R11; wherein one or more methylene linkages of R8and R9are each optionally and independently replaced with an optionally substituted C3-C8 cycloalkylenyl, phenyl, - O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or -C(O)O-; and each R10and R11is independently an optionally substituted cylic, bicyclic, bridged bicyclic, multicyclic or bridged multicyclic C4-C14cycloalkyl or optionally substituted cylic, bicyclic, bridged bicyclic, multicyclic or bridged multicyclic 4-14 membered heterocyclyl, or two R10or two R11taken together form an optionally substituted bridged bicyclic or multicyclic C4-C14 cycloalkyl or optionally substituted bridged bicyclic or multicyclic 4-14 membered heterocyclyl; wherein one or more of X1, XA, X2, X3, X4, X5, X6, X7, R2and R3is optionally and independently substituted with one or more substituents selected from -F, -Cl, -Br and -I. Formula (AX)

[0082] The present disclosure, in some embodiments, provides compounds of Formula (AX)(AX), or a pharmaceutically acceptable salt thereof, wherein: A is selected from an optionally substituted bridged carbocyclic or heterocyclic core selected from thegroup consisting of:n is an integer selected from 1 or 2; R1is selected from the group consisting of -OH, -OAc, -NR2,each R is independently -H or C1-C6 aliphatic; X1and XAare each independently a bond or optionally substituted C1-C6aliphatic; Y1is selected from the group consisting of, and a bond; wherein the bond marked with an "*" is attached to X1; each X2and X3is independently a bond or optionally substituted C1-C12aliphatic; each Y2and Y3is independently selected from the group consisting ofwherein the bond marked with an "*" is attached to X2or X3; each X4and X5is independently optionally substituted C1-C6aliphatic; R2is -CH(OR6)(OR7), -CH(SR6)(SR7), -CH(R6)(R7), -R10, optionally substituted C5-C18aliphatic, or optionally substituted C1-C14 aliphatic-R10, wherein one or more methylene linkages of R2are each optionally and independently replaced with an optionally substituted C3-C8 cycloalkylenyl, phenyl, - O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or -C(O)O-;each R3is independently -CH(OR8)(OR9), -CH(SR8)(SR9), -CH(R8)(R9), -R11, optionally substituted C5-C18 aliphatic, or optionally substituted C1-C14 aliphatic-R11, wherein one or more methylene linkages of R3are each optionally and independently replaced with an optionally substituted C3-C8 cycloalkylenyl, phenyl, -O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or -C(O)O-; R6and R7are each independently optionally substituted -C1-C14aliphatic, -R10, or optionally substituted -C1-C14aliphatic-R10; wherein one or more methylene linkages of R6and R7are each optionally and independently replaced with an optionally substituted C3-C8 cycloalkylenyl, phenyl, - O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or -C(O)O-; R8and R9are each independently optionally substituted -C1-C14 aliphatic, -R11, or optionally substituted -C1-C14 aliphatic-R11; wherein one or more methylene linkages of R8and R9are each optionally and independently replaced with an optionally substituted C3-C8cycloalkylenyl, phenyl, - O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or -C(O)O-; and each R10and R11is independently an optionally substituted cylic, bicyclic, bridged bicyclic, multicyclic or bridged multicyclic C4-C14 cycloalkyl or optionally substituted cylic, bicyclic, bridged bicyclic, multicyclic or bridged multicyclic 4-14 membered heterocyclyl, or two R10or two R11taken together form an optionally substituted bridged bicyclic or multicyclic C4-C14 cycloalkyl or optionally substituted bridged bicyclic or multicyclic 4-14 membered heterocyclyl. Formula (AX-A)

[0083] The present disclosure, in some embodiments, provides compounds of Formula (AX- A)(AX-A), or a pharmaceutically acceptable salt thereof, wherein R1, R, X1, XA, X2, X3, X4, X5, Y1, Y2, Y3, R2, R3, R6, R7, R8, R9, R10, and R11are as described in Formula (AX) or as otherwise described in any embodiments below. Formula (AX-AZ)

[0084] The present disclosure, in some embodiments, provides compounds of Formula (AX- AZ)(AX-AZ), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y1, Y2, Y3, R2, and R3are as described in Formula (AX) or as otherwise described in any embodiments below. Formula (AX-AZ’)

[0085] The present disclosure, in some embodiments, provides compounds of Formula (AX- AZ’)(AX-AZ’), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y1, Y2, Y3, R2, and R3are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-AZ’’)

[0086] The present disclosure, in some embodiments, provides compounds of Formula (AX- AZ’’)(AX-AZ’’), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X4, Y1, Y2, and R2are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-AZ’’’)

[0087] The present disclosure, in some embodiments, provides compounds of Formula (AX- AZ’’’)(AX-AZ’’’), or a pharmaceutically acceptable salt thereof, R1, X1, XA, X2, X4, Y1, Y2, and R2are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below.Formula (AX-AY)

[0088] The present disclosure, in some embodiments, provides compounds of Formula (AX- AY)(AX-AY), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, X6, X7, X8, Y1, Y2, Y3, R2, and R3are as described in Formula (AX’’’) or as otherwise described in any embodiments below. Formula (AX-AY’)

[0089] The present disclosure, in some embodiments, provides compounds of Formula (AX- AZ)(AX-AY’’), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, X6, X7, Y1, Y2, Y3, R2, and R3are as described in Formula (AX’’’) or as otherwise described in any embodiments below. Formula (AX-AY’’)

[0090] The present disclosure, in some embodiments, provides compounds of Formula (AX- AY’’’)(AX-AY’’’), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, X6, X7, Y1, Y2, Y3, R2, and R3are as described in Formulae (AX’’’) or as otherwise described in any embodiments below.Formula (AX-B)

[0091] The present disclosure, in some embodiments, provides compounds of Formula (AX- B)(AX-B), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y1, R2, and R3are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-C)

[0092] The present disclosure, in some embodiments, provides compounds of Formula (AX- C)(AX-C), or a pharmaceutically acceptable salt thereof, R1, X1, XA, X2, X3, X4, X5, Y1, R2, and R3are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-D)

[0093] The present disclosure, in some embodiments, provides compounds of Formula (AX-(AX-D),or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y2, Y3, R2, and R3are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-E), (AX-E’)

[0094] The present disclosure, in some embodiments, provides compounds of Formula (AX- E) or (AX-E’)(AX-E), (AX-E’), or a pharmaceutically acceptable salt thereof, wherein R1, XA, X2, X3, X4, X5, Y2, Y3, R2, and R3are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-F)

[0095] The present disclosure, in some embodiments, provides compounds of Formula (AX- F(AX-F), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y2, Y3, R2, and R3are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-G), (AX-G’)

[0096] The present disclosure, in some embodiments, provides compounds of Formula (AX- G) or (AX-G’)(AX-G), (AX-G’), or a pharmaceutically acceptable salt thereof, wherein R1, XA, X2, X3, X4, X5, Y2, Y3, R2, and R3are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-H), (AX-H’), (AX-H’’), (AX-H’’’), or (AX-H’’’’)

[0097] The present disclosure, in some embodiments, provides compounds of Formula (AX- H), (AX-H’), (AX-H’’), (AX-H’’’), (AX-H’’’’)(AX-H’’’), (AX-H’’’’), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y1, R6, R7, R8, and R9, R10are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below.

[0098] The present disclosure, in some embodiments, provides compounds of Formula (AX- H’’):(AX-H’’), or a pharmaceutically acceptable salt thereof, wherein R6is optionally substituted C4-C10alkyl; R7is optionally substituted C4-C10alkyl; R8is optionally substituted C4-C10 alkyl; R9is optionally substituted C4-C10 alkyl; X4is optionally substituted C1-C6 alkylene; X5is optionally substituted C1-C6alkylene; and XA-R1is selected from the group consisting ofFormula (AX-I), (AX-I’), (AX-I’’), (AX-I’’’), (AX-I’’’’)

[0099] The present disclosure, in some embodiments, provides compounds of Formula (AX- I), (AX-I’), (AX-I’’), (AX-I’’’), or (AX-I’’’’)or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y1, R6, R7, R8, and R9, are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-J), (AX-J’), (AX-J’’), (AX-J’’’), (AX-J’’’’)

[0100] The present disclosure, in some embodiments, provides compounds of Formula (AX- J), (AX-J’), (AX-J’’), (AX-J’’’), or (AX-J’’’’)(AX-J’’’), (AX-J’’’’), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y1, R6, R7, R8, and R9, are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-K), (AX-K’), (AX-K’’), (AX-K’’’), (AX-K’’’’)

[0101] The present disclosure, in some embodiments, provides compounds of Formula (AX- K), (AX-K’), (AX-K’’), (AX-K’’’), or (AX-K’’’’)(AX-K’’’), (AX-K’’’’), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y1, R6, R7, R8, and R9are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-L), (AX-L’), (AX-L’’)

[0102] The present disclosure, in some embodiments, provides compounds of Formula (AX- L), (AX-L’), or (AX-L’’)or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y1, R6, R7, and R11are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-M), (AX-M’), (AX-M’’)

[0103] The present disclosure, in some embodiments, provides compounds of Formula (AX- M), (AX-M’), or (AX-M’’)(AX-M’), (AX-M’’), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y1, R6, R7, and R11are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-N), (AX-N’), (AX-N’’)

[0104] The present disclosure, in some embodiments, provides compounds of Formula (AX- N), (AX-N’), or (AX-N’’)(AX-N’) (AX-N’’), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y1, R8, R9, and R10are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-O), (AX-O’), (AX-O’’)

[0105] The present disclosure, in some embodiments, provides compounds of Formula (AX- O), (AX-O’), or (AX-O’’)(AX-O),(AX-O’) (AX-O’’), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y1, R8, R9, and R10are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-P), (AX-P’), (AX-P’’)

[0106] The present disclosure, in some embodiments, provides compounds of Formula (AX- P), (AX-P’), or (AX-P’’)(AX-P’) (AX-P’’), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y1, R8, R9, and R10are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (A-Q), (AX-Q’), (AX-Q’’)

[0107] The present disclosure, in some embodiments, provides compounds of Formula (AX- Q), (AX-Q’), or (AX-Q’’)(AX-Q’) (AX-Q’’), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y1, R8, R9, and R10are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-R), (AX-R’), (AX-R’’)

[0108] The present disclosure, in some embodiments, provides compounds of Formula (AX- R), (AX-R’), or (AX-R’’)are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below.Formula (AX-S), (AX-S’), (AX-S’’)

[0109] The present disclosure, in some embodiments, provides compounds of Formula (AX- S), (AX-S’), or (AX-S’’)(AX-S’), (AX-S’’), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y1, R6, R7, and R11are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-T), (AX-T’), (AX-T’’)

[0110] The present disclosure, in some embodiments, provides compounds of Formula (AX- T), (AX-T’), or (AX-T’’)(AX-T’), (AX-T’’), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y1, R6, R7, and R11are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-U), (AX-U’), (AX-U’’)

[0111] The present disclosure, in some embodiments, provides compounds of Formula (AX- U), (AX-U’), or (AX-U’’)(AX-U’), (AX-U’’), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y1, R6, R7, R8, and R9are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-V), (AX-V’), (AX-V’’)

[0112] The present disclosure, in some embodiments, provides compounds of Formula (AX- V), (AX-V’), or (AX-V’’)(AX-V),(AX-V’), (AX-V’’), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y1, R6, R7, and R11are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-W), (AX-W’), (AX-W’’)

[0113] The present disclosure, in some embodiments, provides compounds of Formula (AX- W), (AX-W’), or (AX-W’’)(AX-W),(AX-W’), (AX-W’’), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y1, R6, R7, and R11are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-X), (AX-X’), (AX-X’’)

[0114] The present disclosure, in some embodiments, provides compounds of Formula (AX- X), (AX-X’), or (AX-X’’)(AX-X’’), (AX-X’’’),or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y1, R8, R9, and R10are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-Y) (AX-Y’), (AX-Y’’)

[0115] The present disclosure, in some embodiments, provides compounds of Formula (AX- Y) (AX-Y’), or (AX-Y’’),(AX-Y’’), (AX-Y’’’), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y1, R8, R9, and R10are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-Z), (AX-Z’), (AX-Z’’)

[0116] The present disclosure, in some embodiments, provides compounds of Formula (AX- Z), (AX-Z’), or (AX-Z’’)(AX-Z),or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, Y1, R10, and R11are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-AA), (AX-AA’), (AX-AA’’)

[0117] The present disclosure, in some embodiments, provides compounds of Formula (AX- AA), (AX-AA’), or (AX-AA’’)(AX-AA’), (AX-AA’’),or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, Y1, R10, and R11are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-AB), (AX-AB’), (AX-AB’’)

[0118] The present disclosure, in some embodiments, provides compounds of Formula (AX- AB), (AX-AB’), or (AX-AB’’)or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, Y1, R2, and R3are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-AC), (AX-AC’), (AX-AC’’)

[0119] The present disclosure, in some embodiments, provides compounds of Formula (AX- AC), (AX-AC’), or (AX-AC’’)(AX-AC),(AX-AC’), (AX-AC’’), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, Y1, R2, and R3are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-AD), (AX-AD’), (AX-AD’’)

[0120] The present disclosure, in some embodiments, provides compounds of Formula (AX- AD), (AX-AD’), or (AX-AD’’)(AX-AD’), (AX-AD’’), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, Y1, R2, and R11are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below.Formula (AX-AE), (AX-AE’), (AX-AE’’)

[0121] The present disclosure, in some embodiments, provides compounds of Formula (AX- AE), (AX-AE’), or (AX-AE’’)(AX-AE’), (AX-AE’’), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, Y1, R2, and R11are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-AF), (AX-AF’), (AX-AF’’)

[0122] The present disclosure, in some embodiments, provides compounds of Formula (AX- AF), (AX-AF’), or (AX-AF’’)(AX-AF),(AX-AF’), (AX-AF’’), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, Y1, R3, and R10, are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-AG), (AX-AG’), (AX-AG’’)

[0123] The present disclosure, in some embodiments, provides compounds of Formula (AX- AG), (AX-AG’), or (AX-AG’’)(AX-AG’), (AX-AG’’), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, Y1, R3, and R10are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-AH), (AX-AH’), (AX-AH’’), (AX-AH’’’)

[0124] The present disclosure, in some embodiments, provides compounds of Formula (AX- AH), (AX-AH’), (AX-AH’’), or (AX-AH’’’)(AX-AH’’), (AX-AH’’’) or a pharmaceutically acceptable salt thereof, wherein R1, R, X1, XA, X2, X3, X4, X5, Y1, Y2, Y3, R2, R3, R8, R9, and R10are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-AI), (AX-AI’), (AX-AI’’), (AX-AI’’’)

[0125] The present disclosure, in some embodiments, provides compounds of Formula (AX- AI), (AX-AI’), (AX-AI’’), or (AX-AI’’’)(AX-AI’’), (AX-AI’’’) or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y1, Y2, Y3, R2, R3, R8, R9, and R10are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-AJ)

[0126] The present disclosure, in some embodiments, provides compounds of Formula (AX- AJ)(AX-AJ), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y1, R2, and R3are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-AK)

[0127] The present disclosure, in some embodiments, provides compounds of Formula (AX- AK)(AX-AK), or a pharmaceutically acceptable salt thereof, wherein X1, X2, X3, X4, X5, Y2, Y3, R2, and R3are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-AL), (AX-AL’), (AX-AL’’), (AX-AL’’’)

[0128] The present disclosure, in some embodiments, provides compounds of Formula (AX- AL), (AX-AL’), (AX-AL’’), or (AX-AL’’’)(AX-AL’’), (AX-AL’’’),or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y1, R2, R3, R6, R7, R8, R9, and R10are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-AM)

[0129] The present disclosure, in some embodiments, provides compounds of Formula (AX- AM)(AX-AM), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y1, R2, and R3are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-AN)

[0130] The present disclosure, in some embodiments, provides compounds of Formula (AX- AN)(AX-AN), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, R2, and R3are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-AO), (AX-AO’)

[0131] The present disclosure, in some embodiments, provides compounds of Formula (AX- AO) or (AX-AO’)(AX-AO), (AX-AO’) or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, Y1, R3, and R10are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-AX), (AX-AX’)

[0132] The present disclosure, in some embodiments, provides compounds of Formula (AX- AX), or (AX-AX’)(AX-AX), (AX-AX’), or a pharmaceutically acceptable salt thereof, wherein R1, XA, X4, X5, R2, R3, R6, R7, R8, and R9are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-AP), (AX-AP’), (AX-AP’’)

[0133] The present disclosure, in some embodiments, provides compounds of Formula (AX- A ’ ’’(AX-AP),or a pharmaceutically acceptable salt thereof, wherein R1, XA, X4, X5, R2, R3, R6, R7, R8, and R9are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-AQ), (AX-AQ’), (AX-AQ’’)

[0134] The present disclosure, in some embodiments, provides compounds of Formula (AX- A ’ ’’(AX-AQ’), (AX-AQ’’), or a pharmaceutically acceptable salt thereof, wherein R1, XA, X4, X5, R2, R3, R6, R7, R8, and R9are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below.

[0135] The present disclosure, in some embodiments, provides compounds of Formula (AX-A!Q’’):(AX-AQ’’), or a pharmaceutically acceptable salt thereof, wherein R6is optionally substituted C4-C10 alkyl; R7is optionally substituted C4-C10 alkyl; R8is optionally substituted C4-C10 alkyl; R9is optionally substituted C4-C10alkyl; X4is optionally substituted C1-C6alkylene; X5is optionally substituted C1-C6 alkylene; and XA-R1is selected from the group consisting ofFormula (AX-AR), (AX-AR’), (AX-AR’’)

[0136] The present disclosure, in some embodiments, provides compounds of Formula (AX- AR), (AX-AR’), or (AX-AR’’)(AX-AR’’), or a pharmaceutically acceptable salt thereof, wherein R1, XA, X4, X5, R2, R3, R6, R7, R8, and R9are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-AS), (AX-AS’), (AX-AS’’)

[0137] The present disclosure, in some embodiments, provides compounds of Formula (AX- A ’ ’’(AX-AS),or a pharmaceutically acceptable salt thereof, wherein R1, XA, X4, X5, R2, R3, R6, R7, R8, and R9are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-AT), (AX-AT’), (AX-AT’’)

[0138] The present disclosure, in some embodiments, provides compounds of Formula (AX- AT)(AX-AT), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X4, R1, and R2are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below.

[0139] The present disclosure, in some embodiments, provides compounds of Formula (AX- AT’)(AX-AT’),or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, R1, R2, and R3are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below.

[0140] The present disclosure, in some embodiments, provides compounds of Formula (AX- AT’’)(AX-AT’’), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, R1, R2, and R3are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-AW), (AX-AW’), (AX-AW’’)

[0141] The present disclosure, in some embodiments, provides compounds of Formula (AX- AW)(AX-AW), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X4, R1, and R2are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below.

[0142] The present disclosure, in some embodiments, provides compounds of Formula (AX- AW’)(AX-AW’),or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X4, R1, and R2are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below.

[0143] The present disclosure, in some embodiments, provides compounds of Formula (AX- AW’’)(AX-AW’’), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X4, R1, and R2are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AX-AV)

[0144] The present disclosure, in some embodiments, provides compounds of Formula (AX- AV)(AX-AV), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, X6, X7, Y1, Y4, Y5, R2, and R3are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AN)

[0145] The present disclosure, in some embodiments, provides compounds of Formula (AN)(AN), or a pharmaceutically acceptable salt thereof, wherein:A, n, R1, R, X1, XA, X2, X3, X4, X5, Y1, Y2, Y3, R2, R3, R6, R7, R8, R9, R10, and R11are as described in Formulae (AX’’’), (AX’’’’), (AX’’), (AX’) or (AX); each Y4and Y5is independently selected from the group consisting of a bond,; wherein the bond marked with an "*" is attached to X4or X5; and each X6and X7is independently a bond or optionally substituted C1-C6 aliphatic. Formula (AN-A)

[0146] The present disclosure, in some embodiments, provides compounds of Formula (AN- A)(AN-A), or a pharmaceutically acceptable salt thereof, wherein R1X1, XA, X2, X3, X4, X5, X6, X7, Y1, Y4, Y5, R2, and R3are as described in Formulae (AX’’’) or (AN) or as otherwise described in any embodiments below. Formula (AN-B)

[0147] The present disclosure, in some embodiments, provides compounds of Formula (AN- B)(AN-B), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, X7, Y1, Y5, R2, and R3are as described in Formulae (AX’’’) or (AN) or as otherwise described in any embodiments below.Formula (AN-BA), (AN-BA’), (AN-BA’’)

[0148] The present disclosure, in some embodiments, provides compounds of Formula (AN- BA), (AN-BA’), (AN-BA’’)(AN-BA’’), or a pharmaceutically acceptable salt thereof, wherein R1, R2, R3, XA, X4, X5, R6, R7, R8, and R9are as described in Formulae (AX’’’) or (AN) or as otherwise described in any embodiments below. Formula (AN-BB), (AN-BB’), (AN-BB’’)

[0149] The present disclosure, in some embodiments, provides compounds of Formula (AN- BB), (AN-BB’), (AN-BB’’)(AN-BB’’), or a pharmaceutically acceptable salt thereof, wherein R2, R3, XA, X4, X5, R6, R7, R8, and R9are as described in Formulae (AX’’’) or (AN) or as otherwise described in any embodiments below. Formula (AN-BC), (AN-BC’), (AN-BC’’)

[0150] The present disclosure, in some embodiments, provides compounds of Formula (AN- BC), (AN-BC’), (AN-B’’)(AN-BD),(AN-BD’’), or a pharmaceutically acceptable salt thereof, wherein R1, R2, R3, XA, X4, X5, R6, R7, R8, and R9are as described in Formulae (AX’’’) or (AN) or as otherwise described in any embodiments below. described in Formulae (AX’’’) or (AN) or as otherwise described in any embodiments below. Formula (AN-BD), (AN-BD’), (AN-BD’’)

[0151] The present disclosure, in some embodiments, provides compounds of Formula (AN- BD), (AN-BD’), (AN-BD’’)(AN-BC),or a pharmaceutically acceptable salt thereof, wherein R1, R2, R3, XA, X4, X5, R6, R7, R8, and R9are as described in Formulae (AX’’’) or (AN) or as otherwise described in any embodiments below. Formula (AN-BE), (AN-BE’), (AN-BE’’)

[0152] The present disclosure, in some embodiments, provides compounds of Formula (AN- BE), (AN-BE’), (AN-BE’’)(AN-BE’),(AN-BE’’), or a pharmaceutically acceptable salt thereof, wherein R1, R2, R3, XA, X4, X5, R6, R7, R8, and R9are as described in Formulae (AX’’’) or (AN) or as otherwise described in any embodiments below. Formula (AN-C)

[0153] The present disclosure, in some embodiments, provides compounds of Formula (AN- C)(AN-C), or a pharmaceutically acceptable salt thereof, wherein R1, R, X1, XA, X2, X3, X4, X5, X6, X7, Y1, Y4, Y5, R2, R3, R6, R7, R8, R9, R10, and R11are as described in Formulae (AX’’’) or (AN) or as otherwise described in any embodiments below. Formula (AV-A)

[0154] The present disclosure, in some embodiments, provides compounds of Formula (AX- AV)(AX-AV), or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, X6, X7, Y4, Y5, R2, and R3are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AV-AA), (AV-AA’) (AV-AA’’)

[0155] The present disclosure, in some embodiments, provides compounds of Formula (AV- AA), (AV-AA’) (AV-AA’’)(AV-AA’) or a pharmaceutically acceptable salt thereof, wherein R1, X1, XA, X2, X3, X4, X5, R2, R3, R6, R7, R8, and R9are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below.Formula (AV-AB), (AV-AB’) (AV-AB’’)

[0156] The present disclosure, in some embodiments, provides compounds of Formula (AV- AB), (AV-AB’) (AV-AB’’)(AV-AB’) or a pharmaceutically acceptable salt thereof, wherein R1, R, X1, XA, X2, X3, X4, X5, R2, R3, R6, R7, R8, and R9are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AV-AC), (AV-AC’) (AV-AC’’)

[0157] The present disclosure, in some embodiments, provides compounds of Formula (AV- AC), (AV-AC’) (AV-AC’’)(AV-AC’) or a pharmaceutically acceptable salt thereof, wherein R1, R, X1, X2, X3, X4, X5, R2, R3, R6, R7, R8, and R9are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AW)

[0158] The present disclosure, in some embodiments, provides compounds of Formula (AW)(AW), or a pharmaceutically acceptable salt thereof, wherein R1, XA, X2, X3, X4, X5, X6, X7, Y4, Y5, R2, and R3are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AW-AA), (AW-AA’)

[0159] The present disclosure, in some embodiments, provides compounds of Formula (AW- AA), (AW-AA’)(AW-AA’), or a pharmaceutically acceptable salt thereof, wherein R1, XA, X2, X3, X4, X5, R6, R7, R8, and R9are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below.Formula (AW-AB), (AW-AB’)

[0160] The present disclosure, in some embodiments, provides compounds of Formula (AW- AB), (AW-AB’)(AW-AB’), or a pharmaceutically acceptable salt thereof, wherein XA, X2, X3, X4, X5, R6, R7, R8, and R9are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AW-AC), (AW-AC’)

[0161] The present disclosure, in some embodiments, provides compounds of Formula (AW- AC), (AW-AC’)(AW-AC),(AW-AC’), or a pharmaceutically acceptable salt thereof, wherein X4, X5, R6, R7, R8, and R9are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. Formula (AW-AD), (AW-AD’)

[0162] The present disclosure, in some embodiments, provides compounds of Formula (AW- AD), (AW-AD’)(AW-AD’), or a pharmaceutically acceptable salt thereof, wherein R1, XA, R6, R7, R8, and R9are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below.Formula (AW-AE), (AW-AE’)

[0163] The present disclosure, in some embodiments, provides compounds of Formula (AW- AE), (AW-AE’)(AW-AE’), or a pharmaceutically acceptable salt thereof, wherein R1, XA, R6, R7, R8, and R9are as described in Formulae (AX’’’), (AX’’’’), (AX’’) or (AX) or as otherwise described in any embodiments below. A

[0164] As disclosed in Formula (AX’’’’), in certain embodiments, A is selected from an optionally substituted bridged carbocyclic bicycle, bridged carbocyclic multicycle, bridged heterocyclic bicycle, or bridged heterocyclic multicycle. In certain embodiments, A is selected from an optionally substituted 4-16 membered bridged carbocyclic bicycle, bridged carbocyclic multicycle, bridged heterocyclic bicycle, or bridged heterocyclic multicycle. In certain embodiments, A is selected from an optionally substituted 4-14 membered bridged carbocyclic bicycle, bridged carbocyclic multicycle, bridged heterocyclic bicycle, or bridged heterocyclic multicycle. In certain embodiments, A is an optionally substituted 4-16 membered bridged carbocyclic bicycle. In certain embodiments, A is an optionally substituted 4-16 membered bridged carbocyclic multicycle. In certain embodiments, A is an optionally substituted 4-16 membered bridged heterocyclic bicycle. In certain embodiments, A is an optionally substituted 4-16 membered bridged heterocyclic multicycle.In certain embodiments, A is an optionally substituted 4-14 membered bridged carbocyclic bicycle. In certain embodiments, A is an optionally substituted 4-14 membered bridged carbocyclic multicycle. In certain embodiments, A is an optionally substituted 4-14 membered bridged heterocyclic bicycle. In certain embodiments, A is an optionally substituted 4-14 membered bridged heterocyclic multicycle. [

[0166] As disclosed in Formula (AX), in certain embodiments, A is an optionally substitutedembodiments, A isAs disclosed inFormula (AX*), in certain embodiments, A is an optionally substituted core selected from the groupembodiments, A is a multivalent adamantyl core,. In certain embodiments, A isution, as defined in the present disclosure, and the remaining bonds are connected to X1, X2and X3. In certain embodiments, A is a multivalent (1S,5S,7S)-2,4,10-trioxaadamantane core,. In certainembodiments,[eA is, wherein * is attached to X1. In certain embodiments, A is. In certainI Iattached to X1. In certain embodiments, A is. In certain embodiments,certain embodiments, A is ,* attached to X1.In certain embodiments, A is, wherein * is attached to X1 . In certain embodiments,certain embodiments,attached to X1. In certain embodiments, A is , wherein * is attached to X1 . In certain embodiments, A is a multivalent cubane core,embodiments, A is a multivalent bicyclo[2.2.2]octane core,. In certain embodiments, A is cattached to X1. In certain embodiments,certain embodiments, A is amultivalent (1s,4s)-2-oxabicyclo[2.2.2]octane core,. In certain embodiments, A is cattached to X1. In certain embodiments, A is a multivalent (1s,4s)-2-oxabicyclo[2.2.2]octan-3-one core,certain embodiments, A is , wherein * is attached to X1. n and m

[0168] As disclosed in Formula (AX’’), (AX*) and (AX’’’), in certain embodiments, n is an integer selected from 0, 1 or 2 and m is an integer selected from 1 or 2, such that m plus n is less than or equal to 3. In certain embodiments, m is 1 and n is 2. In certain embodiments, m is 2 and n is 1. In certain embodiments, m is 1 and n is 1. In certain embodiments, m is 1 and n is 0. Y

[0169] As disclosed in Formula (AX), in certain embodiments, Y1is,, or bond; wherein the bond marked with an "*" is attached to X1. In certain embodiments,I. In certain embodiments, Y1is. In certain embodiments, Y1is. In certain embodiments, Y1is. In certainembodiments, Y1is. In certain embodiments, Y1is . In certain embodiments,. In certain embodiments, Y1is. In certain embodiments, Y1is a bond.

[0170] In certain embodiments, wherein the compound comprises two Y1(in other words, when m=2); each Y1is independently selected from any of the embodiments above, and need not be the same. In certain embodiments, wherein the compound comprises two Y1, they are each the same. X1

[0171] As disclosed in Formula (AX), in certain embodiments, X1is a bond or optionally substituted C1-C6aliphatic. In certain embodiments, X1is a bond. In certain embodiments, X1is a bond or unsubstituted C1-C6aliphatic. In certain embodiments, X1is unsubstituted C1-C6aliphatic. In certain embodiments, X1is a bond or optionally substituted C1-C6 alkylene. In certain embodiments, X1is optionally substituted C1-C6 alkylene. In certain embodiments, X1is a bond or unsubstituted C1- C6 alkylene. In certain embodiments, X1is unsubstituted C1-C6 alkylene. In certain embodiments, X1is a bond or unsubstituted C1-C4alkylene. In certain embodiments, X1is unsubstituted C1-C4alkylene. In certain embodiments, X1is a bond or unsubstituted C1-C2alkylene. In certain embodiments, X1is unsubstituted C1-C2alkylene. In certain embodiments, X1is unsubstituted C2-C6alkylene. In certain embodiments, X1is optionally substituted methylene. In certain embodiments, R2is optionally substituted C2 alkylene. In certain embodiments, X1is optionally substituted C3 alkylene. In certain embodiments, X1is optionally substituted C4 alkylene. In certain embodiments, X1is optionally substituted C5 alkylene. In certain embodiments, X1is optionally substituted C6 alkylene. In certain embodiments, X1is –(CH2)-. In certain embodiments, X1is –(CH2)2-. In certain embodiments, X1is –(CH2)3-. In certain embodiments, X1is –(CH2)4-. In certain embodiments, X1is –(CH2)5-. In certain embodiments, X1is –(CH2)6-.

[0172] In certain embodiments, X1is substituted with one or more substituents selected from -F, -Cl, -Br and -I. In certain embodiments, X1is substituted with one or more -F. In certain embodiments, X1is substituted with one or more -F on a carbon atom at a position selected from α- position and β-position from Y1.

[0173] In certain embodiments, wherein the compound comprises two X1(in other words, when m=2); each X1is independently selected from any of the embodiments above, and need not be the same. In certain embodiments, wherein the compound comprises two X1, they are each the same. XA

[0174] As disclosed in Formula (AX), in certain embodiments XAis a bond or optionally substituted C1-C6aliphatic. In certain embodiments, XAis a bond. In certain embodiments, XAis a bond or unsubstituted C1-C6aliphatic. In certain embodiments, XAis unsubstituted C1-C6aliphatic. In certain embodiments, XAis a bond or optionally substituted C1-C6 alkylene. In certain embodiments, XAis optionally substituted C1-C6 alkylene. In certain embodiments, XAis a bond or unsubstituted C1- C6 alkylene. In certain embodiments, XAis unsubstituted C1-C6 alkylene. In certain embodiments, XAis a bond or unsubstituted C1-C4alkylene. In certain embodiments, XAis unsubstituted C1-C4alkylene. In certain embodiments, XAis a bond or unsubstituted C1-C2alkylene. In certain embodiments, XAis unsubstituted C1-C2alkylene. In certain embodiments, XAis unsubstituted C2-C6alkylene. In certain embodiments, XAis optionally substituted methylene. In certain embodiments, R2is optionally substituted C2 alkylene. In certain embodiments, XAis optionally substituted C3 alkylene. In certain embodiments, XAis optionally substituted C4 alkylene. In certain embodiments, XAis optionally substituted C5 alkylene. In certain embodiments, XAis optionally substituted C6 alkylene. In certain embodiments, XAis –(CH2)-. In certain embodiments, XAis –(CH2)2-. In certain embodiments, XAis –(CH2)3-. In certain embodiments, XAis –(CH2)4-. In certain embodiments, XAis –(CH2)5-. In certain embodiments, XAis –(CH2)6-.

[0175] In certain embodiments, XAis substituted with one or more substituents selected from -F, -Cl, -Br and -I. In certain embodiments, XAis substituted with one or more -F. In certain embodiments, XAis substituted with one or more -F on a carbon atom at a position selected from α- position and β-position from Y1.

[0176] In certain embodiments, wherein the compound comprises two XA(in other words, when m=2); each XAis independently selected from any of the embodiments above, and need not be the same. In certain embodiments, wherein the compound comprises two XA, they are each the same.

[0177] As disclosed in Formulae (AX) in certain embodiments R1is selected from -OH, -

[0178] As disclosed in Formula (AX’’’), in certain embodiments R1is selected from the group consisting of -OH, -OAc, -NR2, -N(R)RH ,aliphatic-OH and each R is independently -H or C1-C6 aliphatic.

[0179] As disclosed in Formula (AX’’’’), in certain embodiments R1is selected from the His C1-C6 aliphatic-OH and each R is independently -H or C1-C6 aliphatic.

[0180] In certain embodiments, R1is -OH, -NR2, or. In certain embodiments, R1is -NR2or. In certain embodiments, R1is -NMe2, -NEt2, or. In certain embodiments, R1is -OH. In certain embodiments, R1is -OAc. In certain embodiments, R1is -NR2. In certain embodiments, R1is -NH2. In certain embodiments, R1is -NMe2. In certain embodiments, R1is -NEt . In certain12 embodiments, R is . In certain embodiments, R is . In certain embodiments, R1. In certain embodiments,certain embodiments,certain embodiments,certain embodiments, R1is. In certain embodiments, R1. In certain embodiments,certain embodiments, R1is -N(R)RH. In certain embodiments,R1is. In certain embodiments, R1. In certain embodiments, R1is. In certain embodiments, R1is. In certain embodiments, R1is. In certain embodiments,

[0181] In certain embodiments, R1is or comprises -OH or -OAc only A comprises a protonatable nitrogen atom. [ [[

[0185] In certain embodiments, wherein the compound comprises two R1(in other words, when m=2); each R1is independently selected from any of the embodiments above, and need not be the same. In certain embodiments, wherein the compound comprises two R1, they are each the same. R

[0186] As disclosed in Formula (AX), in certain embodiments R is -H or C1-C6 aliphatic. In certain embodiments, R is -H. In certain embodiments, R is C1-C6 aliphatic. In certain embodiments, R is or C1-C6 alkyl. In certain embodiments, R is C1-C4 alkyl. In certain embodiments, R is C1-C2 alkyl. In certain embodiments, R is unsubstituted C1-C6 alkyl. In certain embodiments, R is unsubstituted C1- C4alkyl. In certain embodiments, R is unsubstituted C1-C2alkyl. In certain embodiments, R is methyl or ethyl. X2and X3

[0187] As disclosed in Formula (AX), in certain embodiments, X2and X3are each independently a bond or optionally substituted C1-C12 aliphatic. In certain embodiments, X2and X3are the same. In certain embodiments, X2and X3are different.

[0188] In certain embodiments, X2is a bond. In certain embodiments, X2is an optionally substituted C1-C12alkylene. In certain embodiments, X2is an optionally substituted C1-C12alkenylene. In certain embodiments, X2is an optionally substituted C1-C10aliphatic. In certain embodiments, X2is an optionally substituted C1-C10alkylene. In certain embodiments, X2is an optionally substituted C1- C10 alkenylene. In certain embodiments, X2is an optionally substituted C1-C8 aliphatic. In certainembodiments, X2is an optionally substituted C1-C8 alkylene. In certain embodiments, X2is an optionally substituted C1-C8 alkenylene. In certain embodiments, X2is an optionally substituted C1-C6 aliphatic. In certain embodiments, X2is an optionally substituted C1-C6 alkylene. In certain embodiments, X2is an optionally substituted C1-C6alkenylene. In certain embodiments, X2is an optionally substituted C2-C12aliphatic. In certain embodiments, X2is an optionally substituted C2-C12alkylene. In certain embodiments, X2is an optionally substituted C2-C12alkenylene. In certain embodiments, X2is an optionally substituted C4-C12 aliphatic. In certain embodiments, X2is an optionally substituted C4-C12 alkylene. In certain embodiments, X2is an optionally substituted C4-C12 alkenylene. In certain embodiments, X2is an optionally substituted C4-C10 aliphatic. In certain embodiments, X2is an optionally substituted C4-C10 alkylene. In certain embodiments, X2is an optionally substituted C4-C10alkenylene. In certain embodiments, X2is an optionally substituted C6- C8aliphatic. In certain embodiments, X2is an optionally substituted C6-C8alkylene. In certain embodiments, X2is an optionally substituted C6-C8alkenylene. In certain embodiments, X2is a bond or an optionally substituted C1-C4 aliphatic. In certain embodiments, X2is a bond or an optionally substituted C1-C4 alkylene. In certain embodiments, X2is a bond or an unsubstituted C1-C4 aliphatic. In certain embodiments, X2is a bond or an unsubstituted C1-C4 alkylene. In certain embodiments, X2is a bond or an optionally substituted C1-C2aliphatic. In certain embodiments, X2is a bond or an optionally substituted C1-C2alkylene. In certain embodiments, X2is a bond or an unsubstituted C1-C2aliphatic. In certain embodiments, X2is a bond or an unsubstituted C1-C2alkylene. In certain embodiments, X2is –(CH2)-. In certain embodiments, X2is –(CH2)2-. In certain embodiments, X2is – (CH2)3-. In certain embodiments, X2is –(CH2)4-. In certain embodiments, X2is –(CH2)5-. In certain embodiments, X2is –(CH2)6-. In certain embodiments, X2is –(CH2)7-. In certain embodiments, X2is – (CH2)8-. In certain embodiments, X2is –(CH2)9-. In certain embodiments, X2is –(CH2)10-.

[0189] In certain embodiments, X3is a bond. In certain embodiments, X3is an optionally substituted C1-C12alkylene. In certain embodiments, X3is an optionally substituted C1-C12alkenylene. In certain embodiments, X3is an optionally substituted C1-C10aliphatic. In certain embodiments, X3is an optionally substituted C1-C10 alkylene. In certain embodiments, X3is an optionally substituted C1- C10 alkenylene. In certain embodiments, X3is an optionally substituted C1-C8 aliphatic. In certain embodiments, X3is an optionally substituted C1-C8 alkylene. In certain embodiments, X3is an optionally substituted C1-C8 alkenylene. In certain embodiments, X3is an optionally substituted C1-C6 aliphatic. In certain embodiments, X3is an optionally substituted C1-C6alkylene. In certain embodiments, X3is an optionally substituted C1-C6alkenylene. In certain embodiments, X3is an optionally substituted C2-C12aliphatic. In certain embodiments, X3is an optionally substituted C2-C12alkylene. In certain embodiments, X3is an optionally substituted C2-C12 alkenylene. In certain embodiments, X3is an optionally substituted C4-C12 aliphatic. In certain embodiments, X3is an optionally substituted C4-C12 alkylene. In certain embodiments, X3is an optionally substituted C4-C12 alkenylene. In certain embodiments, X3is an optionally substituted C4-C10 aliphatic. In certainembodiments, X3is an optionally substituted C4-C10 alkylene. In certain embodiments, X3is an optionally substituted C4-C10 alkenylene. In certain embodiments, X3is an optionally substituted C6- C8 aliphatic. In certain embodiments, X3is an optionally substituted C6-C8 alkylene. In certain embodiments, X3is an optionally substituted C6-C8alkenylene. In certain embodiments, X3is a bond or an optionally substituted C1-C4aliphatic. In certain embodiments, X3is a bond or an optionally substituted C1-C4alkylene. In certain embodiments, X3is a bond or an unsubstituted C1-C4aliphatic. In certain embodiments, X3is a bond or an unsubstituted C1-C4 alkylene. In certain embodiments, X3is a bond or an optionally substituted C1-C2 aliphatic. In certain embodiments, X3is a bond or an optionally substituted C1-C2 alkylene. In certain embodiments, X3is a bond or an unsubstituted C1-C2 aliphatic. In certain embodiments, X3is a bond or an unsubstituted C1-C2 alkylene. In certain embodiments, X3is –(CH2)-. In certain embodiments, X3is –(CH2)2-. In certain embodiments, X3is – (CH2)3-. In certain embodiments, X3is –(CH2)4-. In certain embodiments, X3is –(CH2)5-. In certain embodiments, X3is –(CH2)6-. In certain embodiments, X3is –(CH2)7-. In certain embodiments, X3is – (CH2)8-. In certain embodiments, X3is –(CH2)9-. In certain embodiments, X3is –(CH2)10-.

[0190] In certain embodiments, X2and X3are each independently a bond or an optionally substituted C1-C4 aliphatic. In certain embodiments, X2and X3are each independently a bond or an optionally substituted C1-C4alkylene. In certain embodiments, X2and X3are each independently a bond or an unsubstituted C1-C4aliphatic. In certain embodiments, X2and X3are each independently a bond or an unsubstituted C1-C4alkylene. In certain embodiments, X2and X3are each independently a bond or an optionally substituted C1-C2aliphatic. In certain embodiments, X2and X3are each independently a bond or an optionally substituted C1-C2 alkylene. In certain embodiments, X2and X3are each independently a bond or an unsubstituted C1-C2 aliphatic. In certain embodiments, X2and X3are each independently a bond or an unsubstituted C1-C2 alkylene.

[0191] In certain embodiments, X2and X3are both a bond or an optionally substituted C1-C4aliphatic. In certain embodiments, X2and X3are both a bond or an optionally substituted C1-C4alkylene. In certain embodiments, X2and X3are both a bond or an unsubstituted C1-C4aliphatic. In certain embodiments, X2and X3are both a bond or an unsubstituted C1-C4 alkylene. In certain embodiments, X2and X3are both a bond or an optionally substituted C1-C2 aliphatic. In certain embodiments, X2and X3are both a bond or an optionally substituted C1-C2 alkylene. In certain embodiments, X2and X3are both a bond or an unsubstituted C1-C2 aliphatic. In certain embodiments, X2and X3are both a bond or an unsubstituted C1-C2alkylene.

[0192] In certain embodiments, X2and X3are both bonds. In certain embodiments, X2and X3are both –(CH2)-. In certain embodiments, X2and X3are both –(CH2)2-. In certain embodiments, X2and X3are both –(CH2)3-. In certain embodiments, X2and X3are both –(CH2)4-. In certain embodiments, X2and X3are both –(CH2)5-.

[0193] In certain embodiments, X1, X2and X3are all the same. In certain embodiments, X1, X2and X3are all bonds. In certain embodiments, X1, X2and X3are all –(CH2)-. In certainembodiments, X1, X2and X3are all –(CH2)2-. In certain embodiments, X1, X2and X3are all –(CH2)3-. In certain embodiments, X1, X2and X3are all –(CH2)4-. In certain embodiments, X1, X2and X3are all –(CH2)5-.

[0194] In certain embodiments, wherein the compound comprises two X3(in other words, when n=2); each X3is independently selected from any of the embodiments above, and need not be the same. In certain embodiments, wherein the compound comprises two X3, they are each the same.

[0195] In certain embodiments, X2and X3are each independently substituted with one or more substituents selected from -F, -Cl, -Br and -I. In certain embodiments, X2and / or X3are substituted with one or more -F. In certain embodiments, X2and X3are substituted with one or more - F on a carbon atom at a position selected from α-position and β-position from Y2or Y3, respectively. Y2and Y3

[0196] As disclosed in Formula (AX), in certain embodiments, Y2and Y3are each independentlywh2 2 3erein the bond marked with an "*" is attached to X for Y or X for Y3.In certain embodiments, Y2and Y3are the same. In certain embodiments, Y2and Y3are different.

[0197] In certain embodiments, Y2and Y3are each independently, In certain embo2 3diments, Y and Y areeach independentlyIn certain embodiments,Y2and Y3are each independently. In certain embodiments, Y2and Y3are each independently. In certain embodiments, Y2and Y3are each O independently. In certain embodiments, Y2is. In certain embodiments, Y2is. In certain embodiments, Y2is. In certain embodiments,. In certain embodiments, Y2is. In certain embodiments, Y2is. In certain embodiments, Y3is. In certain embodiments, Y3is. In certain embodiments, Y3is. In certain embodiments, Y3is. In certain embodiments, Y3is . In certain embodiments,. In certain embodiments, Y3is. In certain embodiments, Y3and Y2are both. In certain embodiments, Y3and Y2are both. In certain embodiments, Y3and Y2are both. In certain embodiments, Y3and Y2are both.

[0198] In certain embodiments, wherein the compound comprises two Y3(in other words, when n=2); each Y3is independently selected from any of the embodiments above, and need not be the same. In certain embodiments, wherein the compound comprises two Y3, they are each the same. X4and X5

[0199] As disclosed in Formula (AX’’’), in certain embodiments, X4and X5are each independently a bond or optionally substituted C1-C6 aliphatic. As disclosed in Formula (AX), in certain embodiments, X4and X5are each independently optionally substituted C1-C6aliphatic. In certain embodiments, X4and X5are the same. In certain embodiments, X4and X5are different.

[0200] In certain embodiments, X4is a bond. In certain embodiments, X4is an optionally substituted C1-C6 aliphatic. In certain embodiments, X4is an optionally substituted C1-C6 alkylene. In certain embodiments, X4is an optionally substituted C1-C6 alkenylene. In certain embodiments, X4is an optionally substituted C2-C5 aliphatic. In certain embodiments, X4is an optionally substituted C2- C5alkylene. In certain embodiments, X4is an optionally substituted C2-C5alkenylene. In certain embodiments, X4is an optionally substituted C3-C4aliphatic. In certain embodiments, X4is anoptionally substituted C3-C4 alkylene. In certain embodiments, X4is an optionally substituted C3-C4 alkenylene. In certain embodiments, X4is –(CH2)-. In certain embodiments, X4is –(CH2)2-. In certain embodiments, X4is –(CH2)3-. In certain embodiments, X4is –(CH2)4-. In certain embodiments, X4is – (CH2)5-. In certain embodiments, X4is –(CH2)6-.

[0201] In certain embodiments, X5is a bond. In certain embodiments, X5is an optionally substituted C1-C6aliphatic. In certain embodiments, X5is an optionally substituted C1-C6alkylene. In certain embodiments, X5is an optionally substituted C1-C6 alkenylene. In certain embodiments, X5is an optionally substituted C2-C5 aliphatic. In certain embodiments, X5is an optionally substituted C2- C5 alkylene. In certain embodiments, X5is an optionally substituted C2-C5 alkenylene. In certain embodiments, X5is an optionally substituted C3-C4 aliphatic. In certain embodiments, X5is an optionally substituted C3-C4alkylene. In certain embodiments, X5is an optionally substituted C3-C4alkenylene. In certain embodiments, X5is –(CH2)-. In certain embodiments, X5is –(CH2)2-. In certain embodiments, X5is –(CH2)3-. In certain embodiments, X5is –(CH2)4-. In certain embodiments, X5is – (CH2)5-. In certain embodiments, X5is –(CH2)6-.

[0202] In certain embodiments, X4and X5are each independently an optionally substituted C1-C4 aliphatic. In certain embodiments, X4and X5are each independently an optionally substituted C1-C4alkylene. In certain embodiments, X4and X5are each independently an unsubstituted C1-C4aliphatic. In certain embodiments, X4and X5are each independently an unsubstituted C1-C4alkylene. In certain embodiments, X4and X5are each independently an optionally substituted C1-C2aliphatic. In certain embodiments, X4and X5are each independently an optionally substituted C1-C2alkylene. In certain embodiments, X4and X5are each independently an unsubstituted C1-C2 aliphatic. In certain embodiments, X4and X5are each independently an unsubstituted C1-C2 alkylene.

[0203] In certain embodiments, X4and X5are both an optionally substituted C1-C4 aliphatic. In certain embodiments, X4and X5are both an optionally substituted C1-C4alkylene. In certain embodiments, X4and X5are both an unsubstituted C1-C4aliphatic. In certain embodiments, X4and X5are both an unsubstituted C1-C4alkylene. In certain embodiments, X4and X5are both an optionally substituted C1-C2 aliphatic. In certain embodiments, X4and X5are both an optionally substituted C1- C2 alkylene. In certain embodiments, X4and X5are both an unsubstituted C1-C2 aliphatic. In certain embodiments, X4and X5are both an unsubstituted C1-C2 alkylene.

[0204] In certain embodiments, X4and X5are both a bond. In certain embodiments, X4and X5are both –(CH2)-. In certain embodiments, X4and X5are both –(CH2)2-. In certain embodiments, X4and X5are both –(CH2)3-. In certain embodiments, X4is –(CH2)2- and X5is –(CH2)-. In certain embodiments, X4and X5are both –(CH2)4-. In certain embodiments, X4and X5are both –(CH2)5-. In certain embodiments, X4and X5are both –(CH2)6-.

[0205] In certain embodiments, X4and X5are each independently substituted with one or more substituents selected from -F, -Cl, -Br and -I. In certain embodiments, X4and / or X5are substituted with one or more -F. In certain embodiments, X4and X5are substituted with one or more -F on a carbon atom at a position selected from α-position and β-position from Y4or Y5, respectively. In certain embodiments, X4and / or X5are substituted with one or more -F. In certain embodiments, X4and X5are substituted with one or more -F on a carbon atom at a position selected from α-position and β-position from Y2or Y3, respectively. In certain embodiments, one or both of X4and X5are each independently optionally substituted with one or more -CF3. In certain embodiments, one or both of 4X and X5are . In certain embodiments, one or both of X4and X5are. As described in this paragraph, the orientation of the linkage is not intended to be limiting and both orientations / directions are contemplated.

[0206] In certain embodiments, wherein the compound comprises two X5(in other words, when n=2); each X5is independently selected from any of the embodiments above, and need not be the same. In certain embodiments, wherein the compound comprises two X5, they are each the same. Y4and Y5

[0207] As disclosed in Formulae (AN) and (AX’’’), in certain embodiments, Y4and Y5arefor Y4or X5for Y5.In certain embodiments, Y4and Y5are the same. In certain embodiments, Y4and Y5are different.

[0208] In certain embodiments, Y4and Y5are each independently,and Y5are each independently. In certain embodiments, Y4and Y5are each independently. In certain embodiments, Y4and Y5are eachindependently. In certain embodiments, Y4is. In certain. In certain embodiments, Y5is. In certain embodiments, Y5is. In certain embodiments, Y5is. In certain embodiments, Y5is. In certain embodiments, Y5is. In certain embodiments,. In certain embodiments, Y5is. In certain embodiments, Y5and Y4are both. In certain embodiments, Y5and Y4are both. In certain embodiments, Y5and Y4are both. In certain embodiments, Y5and Y4are both.

[0209] In certain embodiments, wherein the compound comprises two Y5(in other words, when n=2); each Y5is independently selected from any of the embodiments above, and need not be the same. In certain embodiments, wherein the compound comprises two Y5, they are each the same. X6and X7

[0210] As disclosed in Formulae (AN) and (AX’’’), in certain embodiments, X6and X7are each independently a bond or optionally substituted C1-C6 aliphatic. In certain embodiments, X6and X7are the same. In certain embodiments, X6and X7are different.

[0211] In certain embodiments, X6is a bond. In certain embodiments, X6is an optionally substituted C1-C6 aliphatic. In certain embodiments, X6is an optionally substituted C1-C6 alkylene. In certain embodiments, X6is an optionally substituted C1-C6 alkenylene. In certain embodiments, X6is an optionally substituted C2-C5aliphatic. In certain embodiments, X6is an optionally substituted C2- C5alkylene. In certain embodiments, X6is an optionally substituted C2-C5alkenylene. In certain embodiments, X6is an optionally substituted C3-C4aliphatic. In certain embodiments, X6is an optionally substituted C3-C4 alkylene. In certain embodiments, X6is an optionally substituted C3-C4 alkenylene. In certain embodiments, X6is –(CH2)-. In certain embodiments, X6is –(CH2)2-. In certain embodiments, X6is –(CH2)3-. In certain embodiments, X6is –(CH2)4-. In certain embodiments, X6is – (CH2)5-. In certain embodiments, X6is –(CH2)6-.

[0212] In certain embodiments, X7is a bond. In certain embodiments, X7is an optionally substituted C1-C6aliphatic. In certain embodiments, X7is an optionally substituted C1-C6alkylene. In certain embodiments, X7is an optionally substituted C1-C6alkenylene. In certain embodiments, X7is an optionally substituted C2-C5 aliphatic. In certain embodiments, X7is an optionally substituted C2- C5 alkylene. In certain embodiments, X7is an optionally substituted C2-C5 alkenylene. In certain embodiments, X7is an optionally substituted C3-C4 aliphatic. In certain embodiments, X7is an optionally substituted C3-C4alkylene. In certain embodiments, X7is an optionally substituted C3-C4alkenylene. In certain embodiments, X7is –(CH2)-. In certain embodiments, X7is –(CH2)2-. In certain embodiments, X7is –(CH2)3-. In certain embodiments, X7is –(CH2)4-. In certain embodiments, X7is – (CH2)5-. In certain embodiments, X7is –(CH2)6-.

[0213] In certain embodiments, X6and X7are each independently an optionally substituted C1-C4 aliphatic. In certain embodiments, X6and X7are each independently an optionally substituted C1-C4 alkylene. In certain embodiments, X6and X7are each independently an unsubstituted C1-C4 aliphatic. In certain embodiments, X6and X7are each independently an unsubstituted C1-C4alkylene. In certain embodiments, X6and X7are each independently an optionally substituted C1-C2aliphatic. In certain embodiments, X6and X7are each independently an optionally substituted C1-C2alkylene. In certain embodiments, X6and X7are each independently an unsubstituted C1-C2 aliphatic. In certain embodiments, X6and X7are each independently an unsubstituted C1-C2 alkylene.

[0214] In certain embodiments, X6and X7are both an optionally substituted C1-C4 aliphatic. In certain embodiments, X6and X7are both an optionally substituted C1-C4 alkylene. In certain embodiments, X6and X7are both an unsubstituted C1-C4aliphatic. In certain embodiments, X6and X7are both an unsubstituted C1-C4alkylene. In certain embodiments, X6and X7are both an optionally substituted C1-C2aliphatic. In certain embodiments, X6and X7are both an optionally substituted C1- C2 alkylene. In certain embodiments, X6and X7are both an unsubstituted C1-C2 aliphatic. In certain embodiments, X6and X7are both an unsubstituted C1-C2 alkylene.

[0215] In certain embodiments, X6and X7are both –(CH2)-. In certain embodiments, X6and X7are both –(CH2)2-. In certain embodiments, X6and X7are both –(CH2)3-. In certain embodiments, X6is a bond and X7is –(CH2)2-. In certain embodiments, X6and X7are both a bond.

[0216] In certain embodiments, X6and X7are each independently substituted with one or more substituents selected from -F, -Cl, -Br and -I. In certain embodiments, X6and / or X7are substituted with one or more -F. In certain embodiments, X6and X7are substituted with one or more - F on a carbon atom at a position selected from α-position and β-position from Y4or Y5, respectively.

[0217] In certain embodiments, wherein the compound comprises two X7(in other words, when n=2); each X7is independently selected from any of the embodiments above, and need not be the same. In certain embodiments, wherein the compound comprises two X7, they are each the same. R2

[0218] As disclosed in Formula (AX), in certain embodiments, R2is -CH(OR6)(OR7), - CH(SR6)(SR7), -CH(R6)(R7), -R10, optionally substituted C5-C18aliphatic, or optionally substituted C1- C14 aliphatic-R10, wherein one or more methylene linkages of R2are each optionally and independently replaced with an optionally substituted C3-C8 cycloalkylenyl, phenyl, -O-, -NH-, -S-, - SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)- or -C(O)O-. In certain embodiments, R2is - CH(OR6)(OR7). In certain embodiments, R2is -CH(R6)(R7). In certain embodiments, R2is - CH(SR6)(SR7). In certain embodiments, R2is -R10. In certain embodiments, R2is optionally substituted C5-C18aliphatic. In certain embodiments, R2is optionally substituted C1-C14aliphatic-R10. In certain embodiments, R2is substituted with one or more -F. As disclosed in Formula (AX’’’), in certain embodiments, R2is -CF(R6)(R7). [[f[

[0222] As disclosed in Formula (AX), in certain embodiments, R3is -CH(OR8)(OR9), - CH(SR8)(SR9), -CH(R8)(R9), -R11, optionally substituted C5-C18aliphatic, or optionally substituted C1- C14 aliphatic-R11, wherein one or more methylene linkages of R3are each optionally and independently replaced with an optionally substituted C3-C8 cycloalkylenyl, phenyl, -O-, -NH-, -S-, - SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)- or -C(O)O-. In certain embodiments, R3is - CH(OR8)(OR9). In certain embodiments, R3is -CH(R8)(R9). In certain embodiments, R3is - CH(SR8)(SR9). In certain embodiments, R3is -R11. In certain embodiments, R3is optionally substituted C5-C18aliphatic. In certain embodiments, R3is optionally substituted C1-C14aliphatic-R11. In certain embodiments, R3is substituted with one or more -F. As disclosed in Formula (AX’’’), in certain embodiments, R3is -CF(R8)(R9). [[f[

[0226] In certain embodiments, R2and R3are the same. In certain embodiments, R2and R3are different.

[0227] As disclosed in Formula (AX), in certain embodiments, R6and R7are each independently optionally substituted -C1-C14 aliphatic, -R10, or optionally substituted -C1-C14 aliphatic-R10; wherein one or more methylene linkages of R6and R7are each optionally and independently replaced with an optionally substituted C3-C8 cycloalkylenyl, phenyl, -O-, -NH-, -S-, - SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)- or -C(O)O-.

[0228] In certain embodiments, R6and R7are the same. In certain embodiments, R6and R7are different.

[0229] In certain embodiments, R6is optionally substituted C1-C14 aliphatic. In certain embodiments, R6is optionally substituted C1-C14 alkyl. In certain embodiments, R6is optionally substituted C1-C14 branched alkyl. In certain embodiments, R6is optionally substituted C1-C14 straight chain alkyl. In certain embodiments, R6is optionally substituted C1-C14 alkenyl. In certain embodiments, R6is optionally substituted C1-C14branched alkenyl. In certain embodiments, R6is optionally substituted C1-C14straight chain alkenyl. In certain embodiments, R6is optionally substituted C4-C10 alkyl. In certain embodiments, R6is optionally substituted straight chain C4-C10 alkyl. In certain embodiments, R6is unsubstituted C6-C10 alkyl. In certain embodiments, R6is unsubstituted straight chain C4-C10 alkyl. In certain embodiments, R6is optionally substituted C6-C10 alkyl. In certain embodiments, R6is optionally substituted straight chain C6-C10 alkyl. In certain embodiments, R6is unsubstituted C4-C10alkyl. In certain embodiments, R6is unsubstituted straight chain C6-C10alkyl. In certain embodiments, R6is optionally substituted –(CH2)5CH3. In certain embodiments, R6is optionally substituted –(CH2)6CH3. In certain embodiments, R6is optionally substituted –(CH2)7CH3. In certain embodiments, R6is optionally substituted –(CH2)8CH3. In certain embodiments, R6is optionally substituted –(CH2)9CH3.

[0230] In certain embodiments, R7is optionally substituted C1-C14 aliphatic. In certain embodiments, R7is optionally substituted C1-C14 alkyl. In certain embodiments, R7is optionally substituted C1-C14 branched alkyl. In certain embodiments, R7is optionally substituted C1-C14 straight chain alkyl. In certain embodiments, R7is optionally substituted C1-C14alkenyl. In certain embodiments, R7is optionally substituted C1-C14branched alkenyl. In certain embodiments, R7is optionally substituted C1-C14straight chain alkenyl. In certain embodiments, R7is optionally substituted C4-C10 alkyl. In certain embodiments, R7is optionally substituted straight chain C4-C10 alkyl. In certain embodiments, R7is unsubstituted C6-C10 alkyl. In certain embodiments, R7is unsubstituted straight chain C4-C10 alkyl. In certain embodiments, R7is optionally substituted C6-C10 alkyl. In certain embodiments, R7is optionally substituted straight chain C6-C10 alkyl. In certain embodiments, R7is unsubstituted C4-C10alkyl. In certain embodiments, R7is unsubstituted straight chain C6-C10alkyl. In certain embodiments, R7is optionally substituted –(CH2)5CH3. In certain embodiments, R7is optionally substituted –(CH2)6CH3. In certain embodiments, R7is optionally substituted –(CH2)7CH3. In certain embodiments, R7is optionally substituted –(CH2)8CH3. In certain embodiments, R7is optionally substituted –(CH2)9CH3.

[0231] In certain embodiments, each R6and R7are each independently selected fromR8and R9

[0232] As disclosed in Formula (AX), in certain embodiments, R8and R9are each independently optionally substituted -C1-C14aliphatic, -R11, or optionally substituted -C1-C14aliphatic-R11; wherein one or more methylene linkages of R8and R9are each optionally and independently replaced with an optionally substituted C3-C8 cycloalkylenyl, phenyl, -O-, -NH-, -S-, - SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)- or -C(O)O-.

[0233] In certain embodiments, R8and R9are the same. In certain embodiments, R8and R9are different.

[0234] In certain embodiments, R8is optionally substituted C1-C14aliphatic. In certain embodiments, R8is optionally substituted C1-C14alkyl. In certain embodiments, R8is optionally substituted C1-C14branched alkyl. In certain embodiments, R8is optionally substituted C1-C14straight chain alkyl. In certain embodiments, R8is optionally substituted C1-C14 alkenyl. In certain embodiments, R8is optionally substituted C1-C14 branched alkenyl. In certain embodiments, R8is optionally substituted C1-C14 straight chain alkenyl. In certain embodiments, R8is optionally substituted C4-C10alkyl. In certain embodiments, R8is optionally substituted straight chain C4-C10alkyl. In certain embodiments, R8is unsubstituted C6-C10alkyl. In certain embodiments, R8isunsubstituted straight chain C4-C10 alkyl. In certain embodiments, R8is optionally substituted C6-C10 alkyl. In certain embodiments, R8is optionally substituted straight chain C6-C10 alkyl. In certain embodiments, R8is unsubstituted C4-C10 alkyl. In certain embodiments, R8is unsubstituted straight chain C6-C10alkyl. In certain embodiments, R8is optionally substituted –(CH2)5CH3. In certain embodiments, R8is optionally substituted –(CH2)6CH3. In certain embodiments, R8is optionally substituted –(CH2)7CH3. In certain embodiments, R8is optionally substituted –(CH2)8CH3. In certain embodiments, R8is optionally substituted –(CH2)9CH3.

[0235] In certain embodiments, R9is optionally substituted C1-C14 aliphatic. In certain embodiments, R9is optionally substituted C1-C14 alkyl. In certain embodiments, R9is optionally substituted C1-C14 branched alkyl. In certain embodiments, R9is optionally substituted C1-C14 straight chain alkyl. In certain embodiments, R9is optionally substituted C1-C14alkenyl. In certain embodiments, R9is optionally substituted C1-C14branched alkenyl. In certain embodiments, R9is optionally substituted C1-C14straight chain alkenyl. In certain embodiments, R9is optionally substituted C4-C10 alkyl. In certain embodiments, R9is optionally substituted straight chain C4-C10 alkyl. In certain embodiments, R9is unsubstituted C6-C10 alkyl. In certain embodiments, R9is unsubstituted straight chain C4-C10 alkyl. In certain embodiments, R9is optionally substituted C6-C10 alkyl. In certain embodiments, R9is optionally substituted straight chain C6-C10alkyl. In certain embodiments, R9is unsubstituted C4-C10alkyl. In certain embodiments, R9is unsubstituted straight chain C6-C10alkyl. In certain embodiments, R9is optionally substituted –(CH2)5CH3. In certain embodiments, R9is optionally substituted –(CH2)6CH3. In certain embodiments, R9is optionally substituted –(CH2)7CH3. In certain embodiments, R9is optionally substituted –(CH2)8CH3. In certain embodiments, R9is optionally substituted –(CH2)9CH3.

[0236] In certain embodiments, each R8and R9are each independently selected from

[0237] In certain embodiments, each R8and R9are each independently selected fromR10and R11

[0238] As disclosed in Formula (AX), in certain embodiments, each R10and R11are independently an optionally substituted cyclic, bicyclic, bridged bicyclic, multicyclic or bridged multicyclic C4-C14cycloalkyl or optionally substituted cylic, bicyclic, bridged bicyclic, multicyclic or bridged multicyclic 4-14 membered heterocyclyl, or two R10or two R11taken together form an optionally substituted bridged bicyclic or multicyclic C4-C14cycloalkyl or optionally substituted bridged bicyclic or multicyclic 4-14 membered heterocyclyl.

[0239] In certain embodiments, each R10and R11are independently an optionally substituted monocyclic C4-C14 cycloalkyl. In certain embodiments, each R10and R11are independently an optionally substituted monocyclic C6-C14 cycloalkyl. In certain embodiments, each R10and R11are independently an optionally substituted monocyclic C6-C8cycloalkyl. In certain embodiments, each R10and R11are independently an optionally substituted bicyclic C4-C14cycloalkyl. In certain embodiments, each R10and R11are independently an optionally substituted bicyclic C6-C14cycloalkyl. In certain embodiments, each R10and R11are independently an optionally substituted bicyclic C8-C14 cycloalkyl. In certain embodiments, each R10and R11are independently an optionally substituted bicyclic C6-C10 cycloalkyl. In certain embodiments, each R10and R11are independently an optionally substituted bridged bicyclic C4-C14cycloalkyl. In certain embodiments, each R10and R11are independently an optionally substituted bridged bicyclic C6-C14cycloalkyl. In certain embodiments, each R10and R11are independently an optionally substituted bridged bicyclic C8-C14cycloalkyl. In certain embodiments, each R10and R11are independently an optionally substituted bridged bicyclic C6-C10 cycloalkyl. In certain embodiments, each R10and R11are independently an optionally substituted multicyclic C4-C14 cycloalkyl. In certain embodiments, each R10and R11are independently an optionally substituted multicyclic C6-C14 cycloalkyl. In certain embodiments, each R10and R11are independently an optionally substituted multicyclic C8-C14cycloalkyl. In certain embodiments, each R10and R11are independently an optionally substituted bridged multicyclic C4-C14cycloalkyl. In certain embodiments, each R10and R11are independently an optionally substituted bridged multicyclic C6-C14 cycloalkyl. In certain embodiments, each R10and R11are independently an optionally substituted bridged multicyclic C8-C14 cycloalkyl. In certain embodiments, two R10or two R11taken together form an optionally substituted bridged bicyclic or multicyclic C4-C14 cycloalkyl. In certain embodiments, two R10or two R11taken together form an optionally substituted bridged bicyclic or multicyclic C6-C14cycloalkyl. In certain embodiments, two R10or two R11taken together form an optionally substituted bridged bicyclic or multicyclic C8-C14cycloalkyl.

[0240] In certain embodiments, each R10and R11are independently an optionally substituted monocyclic 4-14 membered heterocyclyl. In certain embodiments, each R10and R11are independently an optionally substituted monocyclic 6-14 membered heterocyclyl. In certain embodiments, each R10and R11are independently an optionally substituted monocyclic C6-C8 heterocyclyl. In certain embodiments, each R10and R11are independently an optionally substitutedbicyclic 4-14 membered heterocyclyl. In certain embodiments, each R10and R11are independently an optionally substituted bicyclic 6-14 membered heterocyclyl. In certain embodiments, each R10and R11are independently an optionally substituted bicyclic C8-C14 heterocyclyl. In certain embodiments, each R10and R11are independently an optionally substituted bicyclic 6-12 membered heterocyclyl. In certain embodiments, each R10and R11are independently an optionally substituted bridged bicyclic 4- 14 membered heterocyclyl. In certain embodiments, each R10and R11are independently an optionally substituted bridged bicyclic 6-14 membered heterocyclyl. In certain embodiments, each R10and R11are independently an optionally substituted bridged bicyclic C8-C14 heterocyclyl. In certain embodiments, each R10and R11are independently an optionally substituted bridged bicyclic 6-12 membered heterocyclyl. In certain embodiments, each R10and R11are independently an optionally substituted multicyclic 4-14 membered heterocyclyl. In certain embodiments, each R10and R11are independently an optionally substituted multicyclic 6-14 membered heterocyclyl. In certain embodiments, each R10and R11are independently an optionally substituted multicyclic C8-C14heterocyclyl. In certain embodiments, each R10and R11are independently an optionally substituted bridged multicyclic 4-14 membered heterocyclyl. In certain embodiments, each R10and R11are independently an optionally substituted bridged multicyclic 6-14 membered heterocyclyl. In certain embodiments, each R10and R11are independently an optionally substituted bridged multicyclic C8-C14heterocyclyl. In certain embodiments, two R10or two R11taken together form an optionally substituted bridged bicyclic or multicyclic 4-14 membered heterocyclyl. In certain embodiments, two R10or two R11taken together form an optionally substituted bridged bicyclic or multicyclic 6-14 membered heterocyclyl. In certain embodiments, two R10or two R11taken together form an optionally substituted bridged bicyclic or multicyclic 8-14 membered heterocyclyl.

[0241] In certain embodiments, each R10and R11is independently an optionally substituted monovalent cyclic group selected from,embodiments, each R10and R11is independently a structure selected from,i

[0242] In some embodiments, Lipids of the Disclosure are compounds of any one of Formulae (AX), (AX’), (AX”), (AX’’’), (AX’’’’), (AX*) or any of the other Formulae disclosed herein, comprising a ‘core segment’, selected from those of Table (AX-A) below: Table (AX-A). Non-Limiting Examples of Core Segments of Lipids of the DisclosureFormulae (AX), (AX’), (AX”), (AX’’’), (AX’’’’), (AX*) or any of the other Formulae disclosed herein, comprising one or more ‘tail groups’ ,, ,, independently selected from those of Table (AX-B) below: Table (AX-B). Non-Limiting Examples of Tail Segments of Lipids of the Disclosure

[0244] In some embodiments, Lipids of the Disclosure are compounds of any one of Formulae (AX), (AX’), (AX”), (AX’’’), (AX’’’’), (AX*) or any of the other Formulae disclosed herein, comprising one or more ‘head groups’, independently selected from those of Table (AX-C) below: Table (AX-C). Non-Limiting Examples of Head Segments of Lipids of the Disclosure

[0245] In some embodiments, Lipids of the Disclosure are compounds comprising a ‘core segment’ selected from those of Table (AX-A), one or more ‘tail groups’ independently selected from those of Table (AX-B), and one or more ‘head groups’ independently selected from those of Table (AX-C), in all possible permutations and combinations. The present disclosure contemplates each and every compound comprising a combination of core, tail and head segments of Tables (AX-A), (AX- B) and (AX-C), as individual and distinct compounds.

[0246] In some embodiments, Lipids of the Disclosure are selected from any lipid in Table (I) below or a pharmaceutically acceptable salt, solvate, stereoisomer, or enantiomer thereof: Table (I). Non-Limiting Examples of Ionizable Lipids of the Disclosure(I) above, any stereoisomer thereof, or any mixture of stereoisomers thereof, or a pharmaceutically acceptable salt of any of the aforementioned. In some embodiments, Lipids of the Disclosure are selected from any lipid in Table (I) above, an enantiomer thereof, or any mixture of enantiomers thereof, or a pharmaceutically acceptable salt of any of the aforementioned. In some embodiments, Lipids of the Disclosure are selected from any lipid in Table (I) above, any diastereomer thereof, any enantiomer thereof, or any mixture of diastereomers and / or enantiomers thereof, or a pharmaceutically acceptable salt of any of the aforementioned.

[0248] In some embodiments, an LNP of the present disclosure comprises an ionizable lipid selected from a lipid disclosed in PCT Publications WO2023044343A1, WO2023044333A1, WO2023122752A1, WO2024044728A1, and WO2023196931A1, and WO2024192277A1 (each of which is incorporated by reference herein in their entirety). In some embodiments, an ionizable lipid is selected from a lipid disclosed in PCT Publications WO2015095340A1, WO2021021634A1, WO2020237227A1, WO2019236673A1, WO2021226597A1, WO2021113777A1, WO2023056033A1, and WO2023081526A1 (each of which is incorporated by reference herein in their entirety).

[0249] In some embodiments, an LNP of the present disclosure comprises an ionizable lipid disclosed in PCT Application Publication WO2023044343A1, which isincorporated by reference herein, in its entirety. In certain embodiments, the ionizable lipid is one of Formulae (VII-A), (VIII-A), (X), or (X-A) of PCT Application Publication WO2023044343A1.

[0250] In some embodiments, an LNP of the present disclosure comprises an ionizable lipid disclosed in PCT Application Publication WO2023044333A1, which is incorporated by reference herein, in its entirety. In certain embodiments, the ionizable lipid is one of Formulae (CY), (CY-I), (CY-IV), (CY-IV’), or (CY-VI’) of PCT Application Publication WO2023044333A1.

[0251] In some embodiments, an LNP of the present disclosure comprises an ionizable lipid disclosed in PCT Publication WO2023122752A1, which is incorporated by reference herein, in its entirety. In certain embodiments, the ionizable lipid is one of Formulae II, III, VI, VI’, VI’’, VI’’’, VII, VII’, VII’’, VII’’’, VIII, VIII’, VIII’’, VIII’’’, IX, IX’, IX’’, IX’’’, X, X’, X’’, X’’’, XI, XI’, XI’’, XI’’’, XII, XII’, XII’’, XII’’’, XIII, XIII’, XIII’’, XIII’’’, XIV, XIV’, XIV’’, XIV’’’, XV, XV’, XV’’, XV’’’, XVI, XVI’, XVI’’, XVI’’’, XVII, XVIII, XVIII’, XIX, XX, or XXI of PCT Application Publication WO2023122752A1.

[0252] In some embodiments, an LNP of the present disclosure comprises an ionizable lipid disclosed in PCT Publication WO2023196931A1, which is incorporated by reference herein, in its entirety. In certain embodiments, the ionizable lipid is one of Formulae (CX-I), (CX-i), or (CZ) of PCT Application Publication WO2023196931A1. WO2024192277A

[0253] In some embodiments, an LNP of the present disclosure comprises an ionizable lipid disclosed in PCT Publication WO2024192277A1, which is incorporated by reference herein, in its entirety. In certain embodiments, the ionizable lipid is one of Formulae (S-I), (S-M), (AT), (AT-E’), (AT-F’’’), (AT-M), (AT-N’), (AT-O’), (AT-P’’’), (AC), (CO), (CC), (CC-A), (CC-C), (CC-E), (CC-F’), (CC-H), (CC-J), or (CC-L) of PCT Application Publication WO2024192277A1. ii. Structural lipids

[0254] In some embodiments, an LNP comprises a structural lipid. In some embodiments, an LNP comprises two or more structural lipids. Structural lipids can be selected from the group consisting of, but are not limited to, cholesterol, fecosterol, fucosterol, beta sitosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, cholic acid, sitostanol, litocholicacid, tomatine, ursolic acid, alpha-tocopherol, Vitamin D3, Vitamin D2, Calcipotriol, botulin, lupeol, oleanolic acid, beta-sitosterol-acetate and mixtures thereof. In some embodiments, the structural lipid is cholesterol. In some embodiments, the structural lipid is a cholesterol analogue disclosed by Patel, et al., Nat Commun., 11, 983 (2020), which is incorporated herein by reference in its entirety. In some embodiments, the structural lipid comprises cholesterol and a corticosteroid (such as prednisolone, dexamethasone, prednisone, and hydrocortisone), or any combinations thereof. In some embodiments, a structural lipid is described in international patent application WO2019152557A1, which is incorporated herein by reference in its entirety.

[0255] In some embodiments, a structural lipid is a cholesterol analog. Using a cholesterol analog may enhance endosomal escape as described in Patel et al., Naturally-occurring cholesterol analogues in lipid nanoparticles induce polymorphic shape and enhance intracellular delivery of mRNA, Nature Communications (2020), which is incorporated herein by reference.

[0256] In some embodiments, a structural lipid is a phytosterol. Using a phytosterol may enhance endosomal escape as described in Herrera et al., Illuminating endosomal escape of polymorphic lipid nanoparticles that boost mRNA delivery, Biomaterials Science (2020), which is incorporated herein by reference.

[0257] In some embodiments, a structural lipid contains plant sterol mimetics for enhanced endosomal release.

[0258] In some embodiments, the structural lipid is cholesteryl hemisuccinate (CHEMS). In some embodiments, the structural lipid is 3-(4-((2-(4-morpholinyl)ethyl)amino)-4-oxobutanoate) (Mochol). iii. PEGylated lipids

[0259] A PEGylated lipid is a lipid modified with polyethylene glycol. The term “PEGylated lipid” is used interchangeably herein with the shortened term “PEG lipid”.

[0260] In some embodiments, an LNP comprises one, two or more PEGylated lipid or PEG- modified lipid. A PEGylated lipid may be selected from the non-limiting group consisting of PEG- modified phosphatidylethanolamines, PEG-modified phosphatidic acids, PEG-modified ceramides, PEG-modified dialkylamines, PEG-modified diacylglycerols, PEG-modified dialkylglycerols, and mixtures thereof. For example, a PEG lipid may be PEG-c-DOMG, PEG-DMG, PEG-DLPE, PEG- DMPE, PEG-DPPC, or a PEG-DSPE lipid.

[0261] In some embodiments, the PEGylated lipid is selected from (R)-2,3- bis(octadecyloxy)propyl-1-(methoxypoly(ethyleneglycol)2000)propylcarbamate, PEG-S-DSG, PEG- S-DMG, PEG-PE, PEG-PAA, PEG-OH DSPE C18, PEG-DSPE, PEG-DSG, PEG-DPG, PEG- DOMG, PEG-DMPE Na, PEG-DMPE, PEG-DMG2000, PEG-DMG C14, PEG-DMG, PEG-DMA, PEG-Ceramide C16, PEG-C-DOMG, PEG-c-DMOG, PEG-c-DMA, PEG-cDMA, PEGA, PEG750-C- DMA, PEG400, PEG2k-DMG, PEG2k-C11, PEG2000-PE, PEG2000P, PEG2000-DSPE, PEG2000- DOMG, PEG2000-DMG, PEG2000-C-DMA, PEG2000, PEG200, PEG(2k)-DMG, PEG DSPE C18,PEG DMPE C14, PEG DLPE C12, PEG Click DMG C14, PEG Click C12, PEG Click C10, N(Carbonyl-methoxypolyethylenglycol-2000)-l,2-distearoyl-sn-glycero3-phosphoethanolamine, Myrj52, mPEG-PLA, MPEG-DSPE, mPEG3000-DMPE, MPEG-2000-DSPE, MPEG2000-DSPE, mPEG2000-DPPE, mPEG2000-DMPE, mPEG2000-DMG, mDPPE-PEG2000, l,2-distearoyl-sn- glycero-3-phosphoethanolamine-PEG2000, HPEG-2K-LIPD, Folate PEG-DSPE, DSPE-PEGMA 500, DSPE-PEGMA, DSPE-PEG6000, DSPE-PEG5000, DSPE-PEG2K-NAG, DSPE-PEG2k, DSPE- PEG2000maleimide, DSPE-PEG2000, DSPE-PEG, DSG-PEGMA, DSG-PEG5000, DPPE-PEG-2K, DPPE-PEG, DPPE-mPEG2000, DPPE-mPEG, DPG-PEGMA, DOPE-PEG2000, DMPE-PEGMA, DMPE-PEG2000, DMPE-Peg, DMPE-mPEG2000, DMG-PEGMA, DMG-PEG2000, DMG-PEG, distearoyl-glycerol-polyethyleneglycol, C18PEG750, C18PEG5000, C18PEG3000, C18PEG2000, C16PEG2000, C14PEG2000, C18-PEG5000, C18PEG, C16PEG, C16 mPEG (polyethylene glycol) 2000 Ceramide, C14-PEG-DSPE200, C14-PEG2000, C14PEG2000, C14-PEG 2000, C14-PEG, C14PEG, 14:0-PEG2KPE, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-PEG2000, (R)-2,3- bis(octadecyloxy)propyl-1-(methoxypoly(ethyleneglycol)2000)propylcarbamate, (PEG)-C-DOMG, PEG-C-DMA, and DSPE-PEG-X.

[0262] In some embodiments, the LNP comprises a PEGylated lipid disclosed in one of US 2019 / 0240354; US 2010 / 0130588; US 2021 / 0087135; WO 2021 / 204179; US 2021 / 0128488; US 2020 / 0121809; US 2017 / 0119904; US 2013 / 0108685; US 2013 / 0195920; US 2015 / 0005363; US 2014 / 0308304; US 2013 / 0053572; WO 2019 / 232095A1; WO 2021 / 077067; WO 2019 / 152557; US 2015 / 0203446; US 2017 / 0210697; US 2014 / 0200257; or WO 2019 / 089828A1, each of which is incorporated by reference herein in their entirety. In some embodiments, the LNP comprises a PEGylated lipid described or disclosed in PCT / US2023 / 072878 filed August 25, 2023, which is incorporated by reference herein in its entirety.

[0263] In some embodiments, the LNP comprises a PEGylated lipid substitute in place of the PEGylated lipid. All embodiments disclosed herein that contemplate a PEGylated lipid should be understood to also apply to PEGylated lipid substitutes. In some embodiments, the LNP comprises a polysarcosine-lipid conjugate, such as those disclosed in US 2022 / 0001025 A1, which is incorporated by reference herein in its entirety. In some embodiments the LNP comprises a polyoxazoline-lipid conjugate, such as those disclosed in US 2022 / 0249695 A1, which is incorporated by reference herein in its entirety.

[0264] In some embodiments, the LNP comprises a PEGylated lipid disclosed and described in PCT Application WO 2024 / 044728 A1, which is incorporated by reference herein, in its entirety. In certain embodiments, the PEGylated lipid is a lipid of any one of formulas PL-I’, PL-I’’, PL-I, PL- Ia, PL-Ib, PL-Iaa, PL-Iab, PL-Iac, PL-Iad, PL-Iae, PL-Iaf, PL-Iag, PL-Iah, PL-Iba, PL-Ibb, PL-Ibc, PL-Ibd, PL-Ibe, PL-Ibf, PL-Ibg, PL-Ibh, PL-Ica, PL-Icb, PL-Icc, PL-Icd, PL-Id PL-Ie, PL-If, PL-Ig, PL-Ih, PL-Ii, PL-Iha, PL-Ihb, PL-Ihc, PL-Ihd, PL-Iia, PL-Iib, PL-Iic, PL-Iid, PL-Ij, PL-Ik, L-Il, PL- Im, PL-In, PL-Io, PL-Ip, PL-Iq, PL-Ioa, PL-Iob, PL-Ioc, PL-Iod, PL-Ioe, PL-Iof, PL-Iog, PL-Ioh, PL-Ipa, PL-Ipb, PL-Ipc, PL-Ipd, PL-Ipe, PL-Ipf, PL-Ipg, PL-Iph, PL-Iqa, PL-Iqb, PL-Iqc, PL-Iqd, PL-Ir, PL-Is, PL-It, PL-Iu, PL-Iv, PL-Iw, PL-Iva, PL-Ivb, PL-Ivc, PL-Ivd, PL-Iwa, PL-Iwb, PL-Iwc, PL- Iwd, PL-Ix, PL-Ixx, PL-Iy, PL-Iyy, PL-Iyyy, PL-Iz, PL-Izz, PL-Izzz, PL-II’, PL-II’’, PL-II, PL-IIc, PL-IId, PL-IIe, PL-IIf, PL-IIg, PL-IIh, PL-IIa, PL-IIb, PL-IIk, PL-IIm or PL-IIn.

[0265] In some embodiments, the PEGylated lipid is a compound of formula PL-I’:PL-I’ or a pharmaceutically acceptable salt thereof, wherein: A1is a saturated 5-6 membered carbocyclic ring or a saturated 5-6 membered heterocyclic ring containing 1 or 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, wherein the carbocyclic ring and heterocyclic ring are substituted with t occurrences of R4; X1is -N(H)-, -N(C1-6 alkyl)-, -C1-6 aliphatic-N(H)-, -C1-6 aliphatic-N(C1-6 alkyl)-, -O- or -C1-6 aliphatic-O-; L1is -C(O)(C1-6 aliphatic)C(O)-N(R)-, -C(O)(C1-6 aliphatic)-N(R)C(O)-, -C(O)(C1-6 aliphatic)C(O)O-, -C(O)(C1-6aliphatic)C(O)-, -C(O)(C1-6aliphatic)C(O)OCH2-, -C(O)(C1-6aliphatic)-, -C(O)(C1-6aliphatic)-N(R)-, or -C(O)-; L2and L3are independently a covalent bond or C1-6alkylene wherein one methylene unit of the C1-6alkylene is optionally replaced with -O-, -NR-, -S-, -S-S-, -S(O)-, -S(O)2-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)O-, -OC(O)N(R)-, -N(R)C(O)O-, -C(O)N(R)-, -N(R)C(O)-, -N(R)C(O)N(R)-, -C(R5)=N-, or - C(R5)=N-O-; R1is H, C1-6alkyl, -(C1-6alkyl)-N3, -(C1-6alkyl)-SH, or C3-8alkynyl; R2and R3are independently a straight or branched C6-30alkyl, straight or branched C6-30alkenyl, or straight or branched C6-30alkynyl; wherein 1, 2, or 3 methylene units are independently and optionally replaced by a saturated or partially unsaturated C3-6carbocyclic ring or phenylene; wherein the alkyl, alkenyl, and alkynyl and any carbocyclic ring or phenylene is substituted with m instances of Rx; R4is C1-4 alkyl; R5is C1-6 alkyl or C2-14 alkenyl; each R is independently hydrogen or an optionally substituted group selected from C1-6aliphatic, a 3-8 membered saturated or partially unsaturated monocyclic carbocyclic ring, phenyl, an 8-10 membered bicyclic aromatic carbocyclic ring, a 4-8 membered saturated or partially unsaturated monocyclic heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, a 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, or an 8-10 membered bicyclic heteroaromatic ring having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur;each Rxis independently halogen, -CN, -OR, -SR, -C(O)R, -C(O)OR, or -OC(O)OR; n is an integer from 10-75, inclusive; m is 0, 1, 2, 3, or 4; and t is 0, 1, or 2.

[0266] In some embodiments, the PEGylated lipid is a compound of formula PL-II’:PL-II’ or a pharmaceutically acceptable salt thereof, wherein: X1is -N(H)-, -N(C1-6 alkyl)-, -C1-6 aliphatic-N(H)-, -C1-6 aliphatic-N(C1-6 alkyl)-, -O- or -C1-6 aliphatic-O-; L1is -C(O)(C1-6aliphatic)C(O)-, -C(O)(C1-6aliphatic)-, or -C(O)-; L2and L3are a covalent bond or C1-6alkylene wherein one methylene unit of the C1-6alkylene is optionally replaced with -O-, -NR-, -S-, -S-S-, -S(O)-, -S(O)2-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)O-, -OC(O)N(R)-, -N(R)C(O)O-, -C(O)N(R)-, -N(R)C(O)-, -N(R)C(O)N(R)-, -C(R6)=N-, or -C(R6)=N- O-; R1is H, C1-6alkyl, -(C1-6alkyl)-N3, -(C1-6alkyl)-SH, or C3-8alkynyl; R2and R3are independently straight or branched C6-30alkyl, straight or branched C6-30alkenyl, or straight or branched C6-30alkynyl; wherein 1, 2, or 3 methylene units are independently and optionally replaced by a saturated or partially unsaturated C3-6carbocyclic ring or phenylene; wherein the alkyl, alkenyl, and alkynyl and any carbocyclic ring or phenylene is substituted with m instances of Rx; R6is C1-6 alkyl or C2-14 alkenyl; each R is independently hydrogen or an optionally substituted group selected from C1-6 aliphatic, a 3-8 membered saturated or partially unsaturated monocyclic carbocyclic ring, phenyl, an 8-10 membered bicyclic aromatic carbocyclic ring, a 4-8 membered saturated or partially unsaturated monocyclic heterocyclic ring having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, a 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, or an 8-10 membered bicyclic heteroaromatic ring having 1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur; each Rxis independently halogen, -CN, -OR, -SR, -C(O)R, -C(O)OR, or OC(O)OR; n is an integer from 10-75, inclusive; and m is 0, 1, 2, 3, or 4.

[0267] In some embodiments, the PEGylated lipid compound is one of those shown in Table (I-X), or a pharmaceutically acceptable salt thereof.Table (I-X). Exemplary PEGylated Compoundsiv. Phospholipids

[0268] In some embodiments, an LNP of the present disclosure comprises a phospholipid. In some embodiments, an LNP of the present disclosure comprises two or more phospholipids. Phospholipids useful in the compositions and methods may be selected from the non-limiting group consisting of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dioleoyl-sn-glycero-3- phosphoethanolamine (DOPE), 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2- dimyristoyl-sn-glycero-phosphocholine (DMPC), 1.2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-diundecanoyl-sn-glycero-phosphocholine (DUPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn- glycero-3-phosphocholine (18:0 Diether PC), 1-oleoyl-2-cholesterylhemisuc cinoyl-sn-glycero-3- phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2- dilinolenoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2- didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-diphytanoylsn-glycero-3-phosphoethanolamine (ME 16.0 PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn- glycero-3-phosphoethanolamine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1,2- dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG), sodium (S)-2-ammonio-3- ((((R)-2-(oleoyloxy)-3-(stearoyloxy)propoxy)oxidophosphoryl)oxy)propanoate (L-α- phosphatidylserine; Brain PS), dimyristoyl phosphatidylcholine (DMPC), dimyristoyl phosphoethanolamine (DMPE), dimyristoylphosphatidylglycerol (DMPG), dioleoyl- phosphatidylethanolamine4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dioleoylphosphatidylglycerol (DOPG), 1,2-dioleoyl-sn-glycero-3-(phospho-L-serine) (DOPS), acell- fusogenicphospholipid (DPhPE), dipalmitoylphosphatidylethanolamine (DPPE), 1,2-Dielaidoyl-sn- phosphatidylethanolamine (DEPE), dipalmitoylphosphatidylglycerol (DPPG), dipalmitoylphosphatidylserine (DPPS), distearoylphosphatidylcholine (DSPC), distearoyl- phosphatidyl-ethanolamine (DSPE), distearoyl phosphoethanolamineimidazole (DSPEI), 1,2- diundecanoyl-sn-glycero-phosphocholine (DUPC), egg phosphatidylcholine (EPC), 1,2-dioleoyl-sn- glycero-3-phosphate (18:1 PA; DOPA), ammonium bis((S)-2-hydroxy-3-(oleoyloxy)propyl) phosphate (18:1 DMP; LBPA), 1,2-dioleoyl-sn-glycero-3-phospho-(1’-myo-inositol) (DOPI; 18:1 PI), 1,2-distearoyl-sn-glycero-3-phospho-L-serine (18:0 PS), 1,2-dilinoleoyl-sn-glycero-3-phospho-L- serine (18:2 PS), 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine (16:0-18:1 PS; POPS), 1- stearoyl-2-oleoyl-sn-glycero-3-phospho-L-serine (18:0-18:1 PS), 1-stearoyl-2-linoleoyl-sn-glycero-3- phospho-L-serine (18:0-18:2 PS), 1-oleoyl-2-hydroxy-sn-glycero-3-phospho-L-serine (18:1 Lyso PS), 1-stearoyl-2-hydroxy-sn-glycero-3-phospho-L-serine (18:0 Lyso PS), and sphingomyelin. In some embodiments, an LNP comprises DSPC. In certain embodiments, an LNP comprises DOPE. In some embodiments, an LNP comprises both DSPC and DOPE.

[0269] In some embodiments, the LNP comprises a phospholipid selected from 1- pentadecanoyl-2-oleoyl-sn-glycero-3-phosphocholine, 1-myristoyl-2-palmitoyl-sn-glycero-3- phosphocholine, 1-myristoyl-2-stearoyl-sn-glycero-3-phosphocholine, 1-palmitoyl-2-myristoyl-sn- glycero-3-phosphocholine, 1-palmitoyl-2-stearoyl-sn-glycero-3-phosphocholine, 1-palmitoyl-2- oleoyl-glycero-3-phosphocholine, 1-palmitoyl-2-linoleoyl-sn-glycero-3-phosphocholine, 1-palmitoyl- 2-arachidonoyl-sn-glycero-3-phosphocholine, 1-palmitoyl-2-docosahexaenoyl-sn-glycero-3- phosphocholine, 1-stearoyl-2-myristoyl-sn-glycero-3-phosphocholine, 1-stearoyl-2-palmitoyl-sn- glycero-3-phosphocholine, 1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine, 1-stearoyl-2-linoleoyl- sn-glycero-3-phosphocholine, 1-stearoyl-2-arachidonoyl-sn-glycero-3-phosphocholine, 1-stearoyl-2- docosahexaenoyl-sn-glycero-3-phosphocholine, 1-oleoyl-2-myristoyl-sn-glycero-3-phosphocholine, 1-oleoyl-2-palmitoyl-sn-glycero-3-phosphocholine, 1-oleoyl-2-stearoyl-sn-glycero-3-phosphocholine, 1-palmitoyl-2-acetyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-sn-glycero-3-phospho-(1’-myo- inositol-3’,4’-bisphosphate), 1,2-dioleoyl-sn-glycero-3-phospho-(1’-myo-inositol-3’,5’-bisphosphate), 1,2-dioleoyl-sn-glycero-3-phospho-(1’-myo-inositol-4’,5’-bisphosphate), 1,2-dioleoyl-sn-glycero-3- phospho-(1'-myo-inositol-3',4',5'-trisphosphate), 1,2-dioleoyl-sn-glycero-3-phospho-(1’-myo-inositol-3’-phosphate), 1,2-dioleoyl-sn-glycero-3-phospho-(1’-myo-inositol-4’-phosphate), 1,2-dioleoyl-sn- glycero-3-phospho-(1'-myo-inositol-5'-phosphate), 1,2-dioleoyl-sn-glycero-3-phospho-(1’-myo- inositol), 1,2-dioleoyl-sn-glycero-3-phospho-L-serine, and 1-(8Z-octadecenoyl)-2-palmitoyl-sn- glycero-3-phosphocholine.

[0270] In some embodiments, the LNP comprises a phospholipid selected from DSPS (Distearoylphosphatidylserine), DSPG (1,2-distearoyl-sn-glycero-3-phospho-(1'-rac-glycerol)), DSPA (1,2-Distearoyl-sn-glycero-3-phosphate), diPhyPC (1,2-diphytanoyl-sn-glycero-3-phosphocholine), diPhy-diether-PC (1,2-di-O-phytanyl-sn-glycero-3-phosphocholine), diPhyPE (1,2-diphytanoyl-sn- glycero-3-phosphoethanolamine), diPhy-diether-PE (1,2-di-O-phytanyl-sn-glycero-3- phosphoethanolamine), diPhyPS (1,2-diphytanoyl-sn-glycero-3-phospho-L-serine), diPhyPG (1,2- diphytanoyl-sn-glycero-3-phospho-(1'-rac-glycerol)), diPhyPA (1,2-diphytanoyl-sn-glycero-3- phosphate), Egg PA (L-α-phosphatidic acid), and Soy PA (L-α-phosphatidic acid).

[0271] In some embodiments, the LNP comprises a phospholipid selected from 18:1 (Δ9- Cis) PE (DOPE), 18:0-18:1 PE (SOPE), C16-18:1 PE, 16:0-18:1 PE (POPE), 18:1 BMP (S,R), 18:0- 18:1 PC (SOPC), 16:0-18:1 PC (POPC), 4ME 16:0 Diether PE (4Me), 18:1 (Δ9-Trans) PE (DEPE), 16:1 PE (DPPE), and CL. In certain embodiments, the LNP comprises a phospholipid described or disclosed in Alvarez-Benedicto, et al. (Biomater. Sci., 2022, 10, 549) and Li, et al. (Asian Journal of Pharmaceutical Sciences, 2015, 10, 81-98).

[0272] In certain embodiments, the phospholipid is a sphingoid lipid or sphingolipid (used interchangeably herein), such as, but not limited to sphingomyelin. As used herein, the terms “sphingoid lipid” and “sphingolipid” are meant to refer to a class of lipids containing a backbone comprising a sphingoid base. An exemplary sphingoid base is sphingosine. In certain embodiments, the LNP comprises a sphingolipid selected from Egg Sphingomyelin (Egg SM / ESM / (2S,3R,E)-3- hydroxy-2-palmitamidooctadec-4-en-1-yl (2-(trimethylammonio)ethyl) phosphate), Brain or Porcine Sphingomyelin (Brain SM / (2S,3R,E)-3-hydroxy-2-stearamidooctadec-4-en-1-yl (2- (trimethylammonio)ethyl) phosphate), Milk or Bovine Sphingomyelin (Milk SM / (2S,3R,E)-3- hydroxy-2-tricosanamidooctadec-4-en-1-yl (2-(trimethylammonio)ethyl) phosphate), 28:0 SM (N- octacosanoyl-D-erythro-sphingosylphosphorylcholine), 14:0 SM (N-myristoyl-D-erythro- sphingosylphosphorylcholine), 16:1 SM (N-palmitoleoyl-D-erythro-sphingosylphosphorylcholine), 12:0 Dihydro SM (N-lauroyl-D-erythro-sphinganylphosphorylcholine), Lyso SM (Sphingosylphosphorylcholine), Lyso SM (Sphingosylphosphorylcholine), Lyso SM (dihydro) (Sphinganine Phosphorylcholine), 24:1 SM (N-nervonoyl-D-erythro-sphingosylphosphorylcholine), 24:0 SM (N-lignoceroyl-D-erythro-sphingosylphosphorylcholine), 18:1 SM (N-oleoyl-D-erythro- sphingosylphosphorylcholine), 18:0 SM (N-stearoyl-D-erythro-sphingosylphosphorylcholine), 17:0 SM (N-heptadecanoyl-D-erythro-sphingosylphosphorylcholine), 16:0 SM (N-palmitoyl-D-erythro- sphingosylphosphorylcholine), 12:0 SM (N-lauroyl-D-erythro-sphingosylphosphorylcholine), 06:0 SM (N-hexanoyl-D-erythro-sphingosylphosphorylcholine), 02:0 SM (N-acetyl-D-erythro-sphingosylphosphorylcholine), 3-O-methyl Lyso SM (3-O-methyl-spingosylphosphorylcholine), 3-O- methyl-N-methyl Lyso SM (3-O-methyl-N-methyl-spingosylphosphorylcholine), and 3-N-methyl Lyso SM (3-N-methyl-spingosylphosphorylcholine).

[0273] In some embodiments, the LNP comprises a phospholipid comprising at least one constrained tail, such as those described by Gan, et al. (Bioeng Transl Med.2020 Sep; 5(3): e10161.). In certain embodiments, the phospholipid is one selected from:.

[0274] In some embodiments, the LNP comprises a phospholipid comprising a ceramide analogue having a triazole linkage, such as those described by Kim et al., Bioorg. Med. Chem. Lett., 17(16), 2007, 4584-4587.

[0275] In some embodiments, the LNP comprises a phospholipid disclosed in WO 2023 / 141470, which is incorporated by reference herein, in its entirety. In certain embodiments, the

[0276] In some embodiments, the LNP comprises a phospholipid disclosed in WO2022040641A2, which is incorporated by reference herein, in its entirety.

[0277] In some embodiments, a phospholipid tail may be modified in order to promote endosomal escape as described in U.S.2021 / 0121411, which is incorporated herein by reference.

[0278] In some embodiments, the LNP comprises a phospholipid disclosed in one of US 2019 / 0240354; US 2010 / 0130588; US 2021 / 0087135; WO 2021 / 204179; US 2021 / 0128488; US 2020 / 0121809; US 2017 / 0119904; US 2013 / 0108685; US 2013 / 0195920; US 2015 / 0005363; US 2014 / 0308304; US 2013 / 0053572; WO 2019 / 232095A1; WO 2021 / 077067; WO 2019 / 152557; US 2017 / 0210697; or WO 2019 / 089828A1, each of which is incorporated by reference herein in their entirety.

[0279] In some embodiments, phospholipids disclosed in US 2020 / 0121809 have the following structure:wherein R1 and R2 are each independently a branched or straight, saturated or unsaturated carbon chain (e.g., alkyl, alkenyl, alkynyl). v. Targeting moieties

[0280] In some embodiments, the lipid nanoparticle further comprises a targeting moiety. The targeting moiety may be an antibody or a fragment thereof. The targeting moiety may be capable of binding to a target antigen. In certain embodiments, the lipid nanoparticle comprises more than one targeting moiety. In certain embodiments, the lipid nanoparticle comprises more than one targeting moiety, wherein the targeting moieties target at least two different receptors, and in some embodiments, the at least two different receptors are prevalent on different types of cells or tissues.

[0281] In some embodiments, the pharmaceutical composition comprises a targeting moiety that is operably connected to a lipid nanoparticle. In some embodiments, the targeting moiety is capable of binding to a target antigen. In some embodiments, the target antigen is expressed in a target organ. In some embodiments, the target antigen is expressed more in the target organ than it is in the liver.

[0282] In some embodiments, the targeting moiety is an antibody as described in WO2016189532A1, which is incorporated herein by reference. For example, in some embodiments, the targeted particles are conjugated to a specific anti-CD38 monoclonal antibody (mAb), which allows specific delivery of the siRNAs encapsulated within the particles at a greater percentage to B- cell lymphocytes malignancies (such as MCL) than to other subtypes of leukocytes.

[0283] In some embodiments, the targeting moiety targets a receptor selected from CD20, CCR7, CD3, CD4, CD5, CD8, CD16, CD19, CD20, CD21, CD22, CD25, CD28, CD35, CD40, CD45RA, CD45RO, CD52, CD62L, CD80, CD95, CD127, and CD137. In some embodiments, the targeting moiety targets a receptor selected from CD1, CD2, CD3, CD5, CD7, CD8, CD16, CD25, CD26, CD27, CD28, CD30, CD38, CD39, CD40L, CD44, CD45, CD62L, CD69, CD73, CD80, CD83, CD86, CD95, CD103, CD119, CD126, CD150, CD153, CD154, CD161, CD183, CD223, CD254, CD275, CD45RA, CXCR3, CXCR5, FasL, IL18R1, CTLA-4, 0X40, GITR, LAG3, ICOS, PD-1, leu-12, TCR, TLR1, TLR2, TLR3, TLR4, TLR6, NKG2D, CCR, CCR1, CCR2, CCR4, CCR6, and CCR7. In some embodiments, the targeting moiety targets a receptor selected from CD2, CD3, CD5 and CD7. In some embodiments, the targeting moiety targets a receptor selected from CD2,CD3, CD5, CD7, CD8, CD4, beta 7 integrin, beta 2 integrin, and C1q. In some embodiments, the targeting moiety targets CD117. In some embodiments, the targeting moiety targets CD90. In some embodiments, the targeting moiety targets a receptor selected from a mannose receptor, CD206 and C1q. In some embodiments, the targeting moiety is selected from T-cell receptor motif antibodies, T- cell α chain antibodies, T-cell β chain antibodies, T-cell γ chain antibodies, T-cell δ chain antibodies, CCR7 antibodies, CD3 antibodies, CD4 antibodies, CD5 antibodies, CD7 antibodies, CD8 antibodies, CD11b antibodies, CD11c antibodies, CD16 antibodies, CD19 antibodies, CD20 antibodies, CD21 antibodies, CD22 antibodies, CD25 antibodies, CD28 antibodies, CD34 antibodies, CD35 antibodies, CD40 antibodies, CD45RA antibodies, CD45RO antibodies, CD52 antibodies, CD56 antibodies, CD62L antibodies, CD68 antibodies, CD80 antibodies, CD90 antibodies, CD95 antibodies, CD117 antibodies, CD127 antibodies, CD133 antibodies, CD137 (4-1BB) antibodies, CD163 antibodies, F4 / 80 antibodies, IL-4Rα antibodies, Sca-1 antibodies, CTLA-4 antibodies, GITR antibodies GARP antibodies, LAP antibodies, granzyme B antibodies, LFA-1 antibodies, transferrin receptor antibodies, and fragments thereof. In certain embodiments, the targeting moiety is any one described or contemplated in US20230312713A1, US20230203538A1, US20230320995A1, US20160145348, and US20110038941, each of which is incorporated by reference herein in its entirety.

[0284] In some embodiments, the lipid nanoparticles may be targeted when conjugated / attached / associated with a targeting moiety such as an antibody, or a fragment thereof. vi. Zwitterionic amino lipids

[0285] In some embodiments, an LNP comprises a zwitterionic lipid. In some embodiments, an LNP comprising a zwitterionic lipid does not comprise a phospholipid. In some embodiments, an LNP comprising a zwitterionic lipid further comprises a phospholipid.

[0286] Zwitterionic amino lipids have been shown to be able to self-assemble into LNPs without phospholipids to load, stabilize, and release mRNAs intracellularly as described in U.S. Patent Application 20210121411, which is incorporated herein by reference in its entirety. Zwitterionic, ionizable cationic and permanently cationic helper lipids enable tissue-selective mRNA delivery and CRISPR-Cas9 gene editing in spleen, liver and lungs as described in Liu et al., Membrane-destablizing ionizable phospholipids for organ-selective mRNA delivery and CRISPR-Cas gene editing, Nat Mater. (2021), which is incorporated herein by reference in its entirety.

[0287] The zwitterionic lipids may have head groups containing a cationic amine and an anionic carboxylate as described in Walsh et al., Synthesis, Characterization and Evaluation of Ionizable Lysine-Based Lipids for siRNA Delivery, Bioconjug Chem. (2013), which is incorporated herein by reference in its entirety. Ionizable lysine-based lipids containing a lysine head group linked to a long-chain dialkylamine through an amide linkage at the lysine α-amine may reduce immunogenicity as described in Walsh et al., Synthesis, Characterization and Evaluation of Ionizable Lysine-Based Lipids for siRNA Delivery, Bioconjug Chem. (2013).vii. Additional lipid components

[0288] In some embodiments, the LNP compositions of the present disclosure further comprise one or more additional lipid components capable of influencing the tropism of the LNP. In some embodiments, the LNP further comprises at least one lipid selected from DDAB, EPC, 14PA, 18BMP, DODAP, DOTAP, and C12-200 (see Cheng, et al. Nat Nanotechnol.2020 April; 15(4): 313– 320.; Dillard, et al. PNAS 2021 Vol.118 No.52.).

[0289] In some embodiments, an LNP of the present disclosure further comprises one or more additional ionizable lipids, such as, but not limited to those disclosed in one of US 2023 / 0053437; US 2019 / 0240354; US 2010 / 0130588; US 2021 / 0087135; WO 2021 / 204179; US 2021 / 0128488; US 2020 / 0121809; US 2017 / 0119904; US 2013 / 0108685; US 2013 / 0195920; US 2015 / 0005363; US 2014 / 0308304; US 2013 / 0053572; WO 2019 / 232095A1; WO 2021 / 077067; WO 2019 / 152557; US 2017 / 0210697; or WO 2019 / 089828A1, each of which is incorporated by reference herein in their entirety. In certain embodiments, an LNP of the present disclosure further comprises one or more additional ionizable lipids selected from those disclosed in WO2023044343A1 or WO2023044333A1, both of which are incorporated by reference herein in their entirety.

[0290] In some embodiments, the LNP compositions of the present disclosure comprise, or further comprise one or more lipids selected from 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 Diether PC), 1,2-dilinolenoyl-sn-glycero-3-phosphocholine (18:3 PC), Acylcarnosine (AC), 1- hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), N-oleoyl-sphingomyelin (SPM) (C18:l), N- lignoceryl SPM (C24:0), N-nervonoylshphingomyelin (C24:l), Cardiolipin (CL), l,2-bis(tricosa- 10,12-diynoyl)-sn-glycero-3-phosphocholine (DC8-9PC), dicetyl phosphate (DCP), dihexadecyl phosphate (DCP1), 1,2-Dipalmitoylglycerol-3-hemisuccinate (DGSucc), short-chain bis-n- heptadecanoyl phosphatidylcholine (DHPC), dihexadecoyl-phosphoethanolamine (DHPE), 1,2- dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), l,2-dilauroyl-sn-glycero-3-PE (DLPE), dimyristoyl glycerol hemisuccinate (DMGS), dimyristoyl phosphatidylcholine (DMPC), dimyristoyl phosphoethanolamine (DMPE), dimyristoylphosphatidylglycerol (DMPG), dioleyloxybenzylalcohol (DOBA), 1,2-dioleoylglyceryl-3-hemisuccinate (DOGHEMS), N-[2-(2-{2-[2-(2,3-Bis-octadec-9- enyloxy-propoxy)-ethoxy]-ethoxy}-ethoxy)-ethyl]-3-(3,4,5-dihydroxy-6-hydroxymethyl-1etrahydro- pyran-2-ylsulfanyl)-propionamide (DOGP4αMan), dioleoylphosphatidylcholine (DOPC), dioleoylphosphatidylethanolamine (DOPE), dioleoyl-phosphatidylethanolamine4-(N- maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dioleoylphosphatidylglycerol (DOPG), 1,2-dioleoyl-sn-glycero-3-(phospho-L-serine) (DOPS), acell-fusogenicphospholipid (DPhPE), dipalmitoylphosphatidylethanolamine (DPPE), dipalmitoylphosphatidylglycerol (DPPG), dipalmitoylphosphatidylserine (DPPS), distearoylphosphatidylcholine (DSPC), distearoyl- phosphatidyl-ethanolamine (DSPE), distearoyl phosphoethanolamineimidazole (DSPEI), 1,2- diundecanoyl-sn-glycero-phosphocholine (DUPC), egg phosphatidylcholine (EPC), histaminedistearoylglycerol (HDSG), 1,2-Dipalmitoylglycerol-hemisuccinate-Nα-Histidinyl-Hemisuccinate (HistSuccDG), N-(5'-hydroxy-3'-oxypentyl)-10-12-pentacosadiynamide (h-Pegi- PCDA), 2-[l-hexyloxyethyl]-2-devinylpyropheophorbide-a (HPPH), hydrogenatedsoybeanphosphatidylcholine (HSPC), 1,2-Dipalmitoylglycerol-O-α-histidinyl-Nα- hemisuccinate (IsohistsuccDG), mannosialized dipalmitoylphosphatidylethanolamine (ManDOG), l,2- Dioleoyl-sn-Glycero-3-Phosphoethanolamine-N-[4-(p-maleimidomethyl)cyclohexane-carboxamide] (MCC-PE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16:0 PE), 1-myristoyl-2- hydroxy-sn-glycero-phosphocholine (MHPC), a thiol-reactive maleimide headgroup lipid e.g.1,2- dioleoyl-sn-glycero-3-phosphoethanolamine-N-[4-(p-maleimidophenyl)but-yramid (MPB-PE), Nervonic Acid (NA), sodium cholate (NaChol), l,2-dioleoyl-sn-glycero-3-[phosphoethanolamine-N- dodecanoyl (NC12-DOPE), 1-oleoyl-2-cholesteryl hemisuccinoyl-sn-glycero-3-phosphocholine (OChemsPC), phosphatidylethanolamine lipid (PE), PE lipid conjugated with polyethylene glycol(PEG) (e.g., polyethylene glycol-distearoylphosphatidylethanolamine lipid (PEG-PE)), phosphatidylglycerol (PG), partially hydrogenated soy phosphatidylchloline (PHSPC), phosphatidylinositol lipid (PI), phosphotidylinositol-4-phosphate (PIP), palmitoyloleoylphosphatidylcholine (POPC), phosphatidylethanolamine (POPE), palmitoyloleyolphosphatidylglycerol (POPG), phosphatidylserine (PS), lissamine rhodamineB- phosphatidylethanolamine lipid (Rh-PE), purified soy-derived mixture of phospholipids (SIOO), phosphatidylcholine (SM), 18-1-trans-PE,1-stearoyl-2-oleoyl-phosphatidyethanolamine (SOPE), soybean phosphatidylcholine (SPC), sphingomyelins (SPM), alpha,alpha-trehalose-6,6'-dibehenate (TDB), l,2-dielaidoyl-sn-glycero-3-phophoethanolamine (transDOPE), ((23S,5R)-3- (bis(hexadecyloxy)methoxy)-5-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)tetrahydrofuran- 2-yl)methylmethylphosphate, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl- sn-glycero-3-phosphoethanolamine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2- didocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinolenoyl-sn-glycero-3- phosphocholine, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3- phosphoethanolamine, 1,2-dioleyl-sn-glycero-3-phosphoethanolamine, 1,2-distearoyl-sn-glycero-3- phosphoethanolamine, 16-O-monomethyl PE, 16-O-dimethyl PE, and dioleylphosphatidylethanolamine. B. Exemplary LNP Compositions

[0291] In some embodiments, provided herein are LNPs comprising (a): at least one Lipid of the Disclosure; (b) at least one PEG lipid; (c) at least one structural lipid; and (d) at least one non- ionizable lipid and / or a zwitterionic lipid. In some embodiments, the LNPs further comprise an additional ionizable lipid, besides a compound of Formula (AX), or any other formulation disclosed herein.

[0292] In some embodiments, the PEG-lipid is selected from the group consisting of PEG-c- DOMG, PEG-DMG, PEG-DLPE, PEG-DMPE, PEG-DPPC, and PEG-DSPE.

[0293] In some embodiments, the structural lipid is selected from the group consisting of cholesterol, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, and alpha-tocopherol.

[0294] In some embodiments, the non-ionizable lipid is a phospholipid selected from the group consisting of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dioleoyl-sn-glycero-3- phosphoethanolamine (DOPE), 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2- dimyristoyl-sn-glycero-phosphocholine (DMPC), 1.2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-diundecanoyl-sn-glycero-phosphocholine (DUPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocho line (POPC), 1,2-di-O-octadecenyl-sn- glycero-3-phosphocholine (18:0 Diether PC), 1-oleoyl-2-cholesterylhemisuc cinoyl-sn-glycero-3- phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2- dilinolenoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2- didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-diphytanoylsn-glycero-3-phosphoethanolamine (ME 16.0 PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3- phosphoethanolamine, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn- glycero-3-phosphoethanolamine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1,2- dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG), sodium (S)-2-ammonio-3- ((((R)-2-(oleoyloxy)-3-(stearoyloxy)propoxy)oxidophosphoryl)oxy)propanoate (L-α- phosphatidylserine; Brain PS), dimyristoyl phosphatidylcholine (DMPC), dimyristoyl phosphoethanolamine (DMPE), dimyristoylphosphatidylglycerol (DMPG), dioleoyl- phosphatidylethanolamine4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dioleoylphosphatidylglycerol (DOPG), 1,2-dioleoyl-sn-glycero-3-(phospho-L-serine) (DOPS), acell- fusogenicphospholipid (DPhPE), dipalmitoylphosphatidylethanolamine (DPPE), dipalmitoylphosphatidylglycerol (DPPG), dipalmitoylphosphatidylserine (DPPS), distearoylphosphatidylcholine (DSPC), distearoyl-phosphatidyl-ethanolamine (DSPE), distearoyl phosphoethanolamineimidazole (DSPEI), 1,2-diundecanoyl-sn-glycero-phosphocholine (DUPC), egg phosphatidylcholine (EPC), 1,2-dioleoyl-sn-glycero-3-phosphate (18:1 PA; DOPA), ammonium bis((S)-2-hydroxy-3-(oleoyloxy)propyl) phosphate (18:1 DMP; LBPA), 1,2-dioleoyl-sn-glycero-3- phospho-(1’-myo-inositol) (DOPI; 18:1 PI), 1,2-distearoyl-sn-glycero-3-phospho-L-serine (18:0 PS), 1,2-dilinoleoyl-sn-glycero-3-phospho-L-serine (18:2 PS), 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho- L-serine (16:0-18:1 PS; POPS), 1-stearoyl-2-oleoyl-sn-glycero-3-phospho-L-serine (18:0-18:1 PS), 1- stearoyl-2-linoleoyl-sn-glycero-3-phospho-L-serine (18:0-18:2 PS), 1-oleoyl-2-hydroxy-sn-glycero-3- phospho-L-serine (18:1 Lyso PS), 1-stearoyl-2-hydroxy-sn-glycero-3-phospho-L-serine (18:0 Lyso PS), and sphingomyelin.

[0295] In some embodiments, the non-ionizable lipid is a phospholipid selected from the group consisting of Egg Sphingomyelin (Egg SM / ESM / (2S,3R,E)-3-hydroxy-2- palmitamidooctadec-4-en-1-yl (2-(trimethylammonio)ethyl) phosphate), Brain or PorcineSphingomyelin (Brain SM / (2S,3R,E)-3-hydroxy-2-stearamidooctadec-4-en-1-yl (2- (trimethylammonio)ethyl) phosphate), Milk or Bovine Sphingomyelin (Milk SM / (2S,3R,E)-3- hydroxy-2-tricosanamidooctadec-4-en-1-yl (2-(trimethylammonio)ethyl) phosphate), 28:0 SM (N- octacosanoyl-D-erythro-sphingosylphosphorylcholine), 14:0 SM (N-myristoyl-D-erythro- sphingosylphosphorylcholine), 16:1 SM (N-palmitoleoyl-D-erythro-sphingosylphosphorylcholine), 12:0 Dihydro SM (N-lauroyl-D-erythro-sphinganylphosphorylcholine), Lyso SM (Sphingosylphosphorylcholine), Lyso SM (Sphingosylphosphorylcholine), Lyso SM (dihydro) (Sphinganine Phosphorylcholine), 24:1 SM (N-nervonoyl-D-erythro-sphingosylphosphorylcholine), 24:0 SM (N-lignoceroyl-D-erythro-sphingosylphosphorylcholine), 18:1 SM (N-oleoyl-D-erythro- sphingosylphosphorylcholine), 18:0 SM (N-stearoyl-D-erythro-sphingosylphosphorylcholine), 17:0 SM (N-heptadecanoyl-D-erythro-sphingosylphosphorylcholine), 16:0 SM (N-palmitoyl-D-erythro- sphingosylphosphorylcholine), 12:0 SM (N-lauroyl-D-erythro-sphingosylphosphorylcholine), 06:0 SM (N-hexanoyl-D-erythro-sphingosylphosphorylcholine), 02:0 SM (N-acetyl-D-erythro- sphingosylphosphorylcholine), 3-O-methyl Lyso SM (3-O-methyl-spingosylphosphorylcholine), 3-O- methyl-N-methyl Lyso SM (3-O-methyl-N-methyl-spingosylphosphorylcholine), and 3-N-methyl Lyso SM (3-N-methyl-spingosylphosphorylcholine).

[0296] In some embodiments, (a) the PEG lipid is PEG2k-DMG or PEG2k-DSPE or a mixture thereof; (b) the structural lipid is cholesterol; and (c) the phospholipid, non-ionizable lipid or zwitterionic lipid is a sphingolipid or DSPC or a mixture thereof.

[0297] In some embodiments, the lipid component of the nanoparticle comprises: (a) about 0 mol% to about 10 mol% of PEG lipid; (b) about 0 mol% to about 30 mol% structural lipid; (c) about 20 mol% to about 45 mol% phospholipid, non-ionizable lipid or zwitterionic lipid; and (d) about 30 mol% to about 60 mol% of a Lipid of the Disclosure.

[0298] In some embodiments, the lipid component of the nanoparticle comprises: (a) about 1 mol% to about 2 mol% of PEG lipid; (b) about 25 mol% to about 40 mol% structural lipid; (c) about 20 mol% to about 45 mol% phospholipid, non-ionizable lipid or zwitterionic lipid; and (d) about 30 mol% to about 60 mol% of a Lipid of the Disclosure.

[0299] In some embodiments, the lipid component of the nanoparticle comprises: (a) about 2 mol% of PEG lipid; (b) about 25 mol% structural lipid; (c) about 40 mol% phospholipid, non- ionizable lipid or zwitterionic lipid; and (d) about 33 mol% of a Lipid of the Disclosure.

[0300] In some embodiments, the lipid component of the nanoparticle comprises: (a) about 2.5 mol% of PEG lipid; (b) about 39 mol% structural lipid; (c) about 10 mol% phospholipid, non- ionizable lipid or zwitterionic lipid; and (d) about 48.5 mol% of a Lipid of the Disclosure.

[0301] In some embodiments, the lipid component of the nanoparticle comprises: (a) about 1.5 mol% of PEG lipid; (b) about 40 mol% structural lipid; (c) about 10 mol% phospholipid, non- ionizable lipid or zwitterionic lipid; and (d) about 48.5 mol% of a Lipid of the Disclosure.

[0302] In certain embodiments, the lipid component of the nanoparticle composition comprises about 30 mol % to about 60 mol % ionizable lipid, about 0 mol % to about 30 mol % phospholipid, about 18.5 mol % to about 48.5 mol % structural lipid, and about 0 mol% to about 10 mol% of PEG lipid, provided that the total mol % does not exceed 100%. In certain embodiments, the lipid component of the nanoparticle composition comprises about 20 mol % to about 45 mol % ionizable lipid, about 30 mol % to about 60 mol % phospholipid, about 10 mol % to about 30 mol % structural lipid, and about 0 mol% to about 10 mol% of PEG lipid, provided that the total mol % does not exceed 100%. In some embodiments, the lipid component of the nanoparticle composition comprises about 35 mol % to about 55 mol % ionizable lipid, about 5 mol % to about 25 mol % phospholipid, about 30 mol % to about 40 mol % structural lipid, and about 0 mol % to about 10 mol % of PEG lipid, provided that the total mol % does not exceed 100%. In some embodiments, the lipid component of the nanoparticle composition comprises about 30 mol % to about 40 mol % ionizable lipid, about 35 mol % to about 45 mol % phospholipid, about 20 mol % to about 30 mol % structural lipid, and about 0.5 mol % to about 5 mol % of PEG lipid, provided that the total mol % does not exceed 100%. In certain embodiments, the lipid component of the nanoparticle composition comprises about 25 mol % to about 45 mol % ionizable lipid, about 35 mol % to about 50 mol % phospholipid, about 10 mol % to about 25 mol % structural lipid, and about 1 mol% to about 5 mol% of PEG lipid, provided that the total mol % does not exceed 100%. In a particular embodiment, the lipid component comprises about 50 mol % ionizable lipid, about 10 mol % phospholipid, about 38.5 mol % structural lipid, and about 1.5 mol% of PEG lipid. In another particular embodiment, the lipid component comprises about 40 mol % ionizable lipid, about 20 mol % phospholipid, about 38.5 mol % structural lipid, and about 1.5 mol % of PEG lipid. In another particular embodiment, the lipid component comprises about 48.5 mol % ionizable lipid, about 10 mol % phospholipid, about 40 mol % structural lipid, and about 1.5 mol % of PEG lipid. In another particular embodiment, the lipid component comprises about 48.5 mol % ionizable lipid, about 10 mol % phospholipid, about 39 mol % structural lipid, and about 2.5 mol % of PEG lipid. In another particular embodiment, the lipid component comprises about 33 mol % ionizable lipid, about 40 mol % phospholipid, about 25 mol % structural lipid, and about 2 mol % of PEG lipid. In some embodiments, the phospholipid is DOPE or DSPC. In some embodiments, the phospholipid is DSPC. In some embodiments, the phospholipid is a sphingolipid. In some embodiments, the phospholipid is a sphingomyelin. In other embodiments, the PEG lipid is PEG-DMG (eg. PEG2K-DMG). In other embodiments, the PEG lipid is PEG-DSPE (eg. PEG2K-DSPE). In other embodiments, the PEG lipid is PEG-DMPE (eg. PEG2K-DMPE). In other embodiments, the structural lipid is cholesterol. In other embodiments, the PEG lipid is PEG- DMG and / or the structural lipid is cholesterol. In some embodiments, the PEG lipids is PEG2K- DMG, the structural lipid is cholesterol, and the phospholipid is DSPC. In some embodiments, the PEG lipids is PEG2K-DMG, the structural lipid is cholesterol, and the phospholipid is sphingomyelin. In some embodiments, the PEG lipids is PEG-DMG, the structural lipid is cholesterol, and thephospholipid is a mixture of DSPC and sphingomyelin. In certain embodiments, the LNP comprises about 33mol% ionizable lipid (eg. at least one ionizable lipid of a Formula described herein), about 40mol% of a sphingolipid, about 25mol% cholesterol and about 2mol% PEG2K-DMG. In some embodiments, the PEG lipids is PEG2K-DSPE, the structural lipid is cholesterol, and the phospholipid is DSPC. In some embodiments, the PEG lipids is PEG2K-DSPE, the structural lipid is cholesterol, and the phospholipid is sphingomyelin. In some embodiments, the PEG lipids is PEG- DSPE, the structural lipid is cholesterol, and the phospholipid is a mixture of DSPC and sphingomyelin. In some embodiments, the PEG lipids is PEG2K-DMG, the structural lipid is cholesterol, and the phospholipid is DOPE. In some embodiments, the PEG lipids is PEG2K-DMG, the structural lipid is cholesterol, and the phospholipid is DOPC. In some embodiments, the PEG lipids is PEG2K-DMG, the structural lipid is cholesterol, and the phospholipid is DLPC. In some embodiments, the PEG lipids is PEG2K-DMG, the structural lipid is cholesterol, and the phospholipid is DOPS. In some embodiments, the PEG lipids is PEG-DMG, the structural lipid is cholesterol, and the phospholipid is a mixture of a phosphatidylcholine lipid and a sphingolipid. In some embodiments, the PEG lipids is PEG-DMG, the structural lipid is cholesterol, and the phospholipid is a mixture of a phosphatidylcholine lipid and phosphatidylserine lipid. In some embodiments, the PEG lipids is PEG-DMG, the structural lipid is cholesterol, and the phospholipid is a mixture of a phosphatidylcholine lipid and a phosphoethanolamine lipid. In some embodiments, the PEG lipids is PEG-DMG, the structural lipid is cholesterol, and the phospholipid is a mixture of a sphingolipid and phosphatidylserine lipid. In some embodiments, the PEG lipids is PEG-DMG, the structural lipid is cholesterol, and the phospholipid is a mixture of a sphingolipid and a phosphoethanolamine lipid. In certain embodiments, the LNP comprises about 33mol% ionizable lipid, about 20mol% of a sphingolipid, about 20mol% of a non-sphingolipid phospholipid, about 25mol% cholesterol and about 2mol% of a PEGylated lipid. In certain embodiments, the LNP comprises about 33mol% ionizable lipid, about 10mol% of a sphingolipid, about 30mol% of a non-sphingolipid phospholipid, about 25mol% cholesterol and about 2mol% of a PEGylated lipid. In certain embodiments, the LNP comprises about 33mol% ionizable lipid, about 30mol% of a sphingolipid, about 10mol% of a non- sphingolipid phospholipid, about 25mol% cholesterol and about 2mol% of a PEGylated lipid. In certain embodiments, the LNP comprises about 33mol% ionizable lipid, about 20mol% sphingomyelin, about 20mol% of a DSPC, about 25mol% cholesterol and about 2mol% of a PEGylated lipid. In certain embodiments, the LNP comprises about 33mol% ionizable lipid, about 10mol% sphingomyelin, about 30mol% of a DSPC, about 25mol% cholesterol and about 2mol% of a PEGylated lipid. In certain embodiments, the LNP comprises about 33mol% ionizable lipid, about 30mol% sphingomyelin, about 10mol% of a DSPC, about 25mol% cholesterol and about 2mol% of a PEGylated lipid. In certain embodiments, the LNP comprises about 33mol% ionizable lipid, about 25mol% cholesterol, about 2mol% of a PEGylated lipid, and about 40% of a mixture of phosphatidylcholine, phosphatidylserine, phosphoethanolamine, and sphingoid lipids. In certainembodiments, the LNP comprises about 33mol% ionizable lipid, about 25mol% cholesterol, about 2mol% of a PEGylated lipid, and about 40% of a mixture of phosphatidylcholine, phosphatidylserine, phosphoethanolamine, and sphingoid lipids, wherein each of the phosphatidylcholine, phosphatidylserine, phosphoethanolamine, and sphingoid lipids is present in an amount less than 30 mol% of the total lipid component of the LNP. In certain embodiments, the LNP comprises about 33mol% ionizable lipid, about 25mol% cholesterol, about 2mol% of a PEGylated lipid, and about 40% of a mixture of phosphatidylcholine, phosphatidylserine, phosphoethanolamine, and / or sphingoid lipids, wherein each of the phosphatidylcholine, phosphatidylserine, phosphoethanolamine, and / or sphingoid lipids is present in an amount less than 25 mol% of the total lipid component of the LNP. In certain embodiments, LNP is any one of the aforementioned in this paragraph wherein the PEG lipid is PEG2k-DMG. In certain embodiments, LNP is any one of the aforementioned in this paragraph wherein the PEG lipid is PEG2k-DSPE.

[0303] In another particular embodiment, LNP comprises about 33 mol % ionizable lipid, about 40 mol % DSPC, about 25 mol % cholesterol, and about 2 mol % of PEG lipid. In another particular embodiment, LNP comprises about 33 mol % ionizable lipid, about 40 mol % sphingomyelin, about 25 mol % cholesterol, and about 2 mol % of PEG lipid. In another particular embodiment, LNP comprises about 33 mol % ionizable lipid, about 40 mol % DOPE, about 25 mol % cholesterol, and about 2 mol % of PEG lipid. In another particular embodiment, LNP comprises about 33 mol % ionizable lipid, about 40 mol % DOPC, about 25 mol % cholesterol, and about 2 mol % of PEG lipid. In another particular embodiment, LNP comprises about 33 mol % ionizable lipid, about 40 mol % DLPC, about 25 mol % cholesterol, and about 2 mol % of PEG lipid. In another particular embodiment, LNP comprises about 33 mol % ionizable lipid, about 40 mol % DOPS, about 25 mol % cholesterol, and about 2 mol % of PEG lipid. In another particular embodiment, LNP comprises about 33 mol % ionizable lipid, about 40 mol % phospholipid, about 25 mol % cholesterol, and about 2 mol % of PEG lipid. In another particular embodiment, LNP comprises about 33 mol % ionizable lipid, about 20 mol % sphingomyelin, about 20 mol% DSPC, about 25 mol % cholesterol, and about 2 mol % of PEG lipid. In certain embodiments, LNP is any one of the aforementioned in this paragraph wherein the PEG lipid is PEG2k-DMG. In certain embodiments, LNP is any one of the aforementioned in this paragraph wherein the PEG lipid is PEG2k-DSPE.

[0304] In certain embodiments, the LNP comprises about 43mol% ionizable lipid, about 15mol% of a sphingolipid, about 15mol% of a non-sphingolipid phospholipid, about 25mol% cholesterol and about 2mol% of a PEGylated lipid. In certain embodiments, the LNP comprises about 33mol% ionizable lipid, about 25mol% of a sphingolipid, about 15mol% of a non-sphingolipid phospholipid, about 25mol% cholesterol and about 2mol% of a PEGylated lipid. In certain embodiments, the LNP comprises about 33mol% ionizable lipid, about 15mol% of a sphingolipid, about 25mol% of a non-sphingolipid phospholipid, about 25mol% cholesterol and about 2mol% of a PEGylated lipid. In some embodiments, the PEG lipid is PEG2K-DSPE, the structural lipid ischolesterol, and the phospholipid is a mixture of DSPC and sphingomyelin. In some embodiments, the PEG lipid is PEG2K-DMG, the structural lipid is cholesterol, and the phospholipid is a mixture of DSPC and sphingomyelin. In certain embodiments, the LNP comprises about 48.5mol% ionizable lipid, about 10mol% of a phospholipid (such as DSPC), about 40mol% cholesterol and about 1.5mol% PEG2K-DSPE. In certain embodiments, the LNP comprises about 48.5mol% ionizable lipid, about 10mol% of a phospholipid (such as DSPC), about 40mol% cholesterol and about 1.5mol% PEG2K-DMG. In certain embodiments, the LNP comprises about 48.5mol% ionizable lipid, about 10mol% of a phospholipid (such as DSPC), about 39mol% cholesterol and about 2.5mol% PEG2K- DSPE.

[0305] In another particular embodiment, the lipid component includes about 48.5 mol % ionizable lipid, about 10 mol % phospholipid, about 38.5 mol % structural lipid, and about 3 mol % of PEG lipid. In another particular embodiment, the lipid component includes about 48.5 mol % ionizable lipid, about 10 mol % phospholipid, about 38 mol % structural lipid, and about 3.5 mol % of PEG lipid. In some embodiments, the PEG lipid is PEG2K-DPPE, the structural lipid is cholesterol, and the phospholipid is a DSPC or a mixture of DSPC and sphingomyelin. In some embodiments, the PEG lipid is PEG2K-DPPE, the structural lipid is cholesterol, and the phospholipid is a mixture of DSPC and sphingomyelin. In certain embodiments, the LNP comprises about 48.5mol% ionizable lipid, about 10mol% of a phospholipid (such as DSPC), about 40mol% cholesterol and about 1.5mol% PEG2K-DPPE. In certain embodiments, the LNP comprises about 48.5mol% ionizable lipid, about 10mol% of a phospholipid (such as DSPC), about 39.5 mol% cholesterol and about 2 mol% PEG2K- DPPE. In certain embodiments, the LNP comprises about 48.5mol% ionizable lipid, about 10mol% of a phospholipid (such as DSPC), about 39mol% cholesterol and about 2.5mol% PEG2K-DPPE. In certain embodiments, the LNP comprises about 48.5mol% ionizable lipid, about 10mol% of a phospholipid (such as DSPC), about 38.5 mol% cholesterol and about 3 mol% PEG2K-DPPE. In certain embodiments, the LNP comprises about 48.5mol% ionizable lipid, about 10mol% of a phospholipid (such as DSPC), about 38 mol% cholesterol and about 3.5mol% PEG2K-DPPE. In some embodiments, the PEG lipid is PEG2K-DMG, the structural lipid is cholesterol, and the phospholipid is a DSPC or a mixture of DSPC and sphingomyelin. In some embodiments, the PEG lipid is PEG2K- DMG, the structural lipid is cholesterol, and the phospholipid is a mixture of DSPC and sphingomyelin. In certain embodiments, the LNP comprises about 48.5mol% ionizable lipid, about 10mol% of a phospholipid (such as DSPC), about 40mol% cholesterol and about 1.5mol% PEG2K- DMG. In certain embodiments, the LNP comprises about 48.5mol% ionizable lipid, about 10mol% of a phospholipid (such as DSPC), about 39.5 mol% cholesterol and about 2 mol% PEG2K-DMG. In certain embodiments, the LNP comprises about 48.5mol% ionizable lipid, about 10mol% of a phospholipid (such as DSPC), about 39mol% cholesterol and about 2.5mol% PEG2K-DMG. In certain embodiments, the LNP comprises about 48.5mol% ionizable lipid, about 10mol% of a phospholipid (such as DSPC), about 38.5 mol% cholesterol and about 3 mol% PEG2K-DMG. Incertain embodiments, the LNP comprises about 48.5mol% ionizable lipid, about 10mol% of a phospholipid (such as DSPC), about 38 mol% cholesterol and about 3.5mol% PEG2K-DMG. In some embodiments, the PEG lipid is PEG2K-DSPE, the structural lipid is cholesterol, and the phospholipid is a DSPC or a mixture of DSPC and sphingomyelin. In some embodiments, the PEG lipid is PEG2K- DSPE, the structural lipid is cholesterol, and the phospholipid is a mixture of DSPC and sphingomyelin. In certain embodiments, the LNP comprises about 48.5mol% ionizable lipid, about 10mol% of a phospholipid (such as DSPC), about 40mol% cholesterol and about 1.5mol% PEG2K- DSPE. In certain embodiments, the LNP comprises about 48.5mol% ionizable lipid, about 10mol% of a phospholipid (such as DSPC), about 39.5 mol% cholesterol and about 2 mol% PEG2K-DSPE. In certain embodiments, the LNP comprises about 48.5mol% ionizable lipid, about 10mol% of a phospholipid (such as DSPC), about 39mol% cholesterol and about 2.5mol% PEG2K-DSPE. In certain embodiments, the LNP comprises about 48.5mol% ionizable lipid, about 10mol% of a phospholipid (such as DSPC), about 38.5 mol% cholesterol and about 3 mol% PEG2K-DSPE. In certain embodiments, the LNP comprises about 48.5mol% ionizable lipid, about 10mol% of a phospholipid (such as DSPC), about 38 mol% cholesterol and about 3.5mol% PEG2K-DSPE.

[0306] In some embodiments, the LNP further comprises a targeting moiety. In some embodiments, the targeting moiety is an antibody or a fragment thereof.

[0307] In some embodiments, the LNP further comprises an active agent. In some embodiments, the active agent is a nucleic acid. In some embodiments, the nucleic acid is a ribonucleic acid. In some embodiments, the ribonucleic acid is at least one ribonucleic acid selected from the group consisting of a small interfering RNA (siRNA), an asymmetrical interfering RNA (aiRNA), a microRNA (miRNA), a Dicer-substrate RNA (dsRNA), a small hairpin RNA (shRNA), a messenger RNA (mRNA), and a long non-coding RNA (lncRNA). In some embodiments, the nucleic acid is a messenger RNA (mRNA) or a circular RNA. In some embodiments, the mRNA comprises an open reading frame encoding a cancer antigen. In some embodiments, the mRNA comprises an open reading frame encoding an immune checkpoint modulator. In some embodiments, the mRNA comprises at least one motif selected from the group consisting of a stem loop, a chain terminating nucleoside, a polyA sequence, a polyadenylation signal, and a 5' cap structure. In some embodiments, the nucleic acid is suitable for a genome editing technique. In some embodiments, the genome editing technique is clustered regularly interspaced short palindromic repeats (CRISPR) or transcription activator-like effector nuclease (TALEN). In some embodiments, the nucleic acid is at least one nucleic acid suitable for a genome editing technique selected from the group consisting of a CRISPR RNA (crRNA), a trans-activating crRNA (tracrRNA), a single guide RNA (sgRNA), and a DNA repair template. In some embodiments, the mRNA is at least 30 nucleotides in length. In some embodiments, the mRNA is at least 300 nucleotides in length. In some embodiments, the nucleic acid encodes a therapeutic protein. In some embodiments, the therapeutic protein is a CAR or TCR complex protein. In some embodiments, the CAR or TCR complex protein comprises an antigenbinding domain specific for an antigen selected from the group: CD 19, CD123, CD22, CD30, CD171, CS-1, C-type lectin-like molecule- 1, CD33, epidermal growth factor receptor variant III (EGFRvIII), disialoganglioside GD2, disaloganglioside GD3, TNF receptor family member, B cell maturation antigen (BCMA), Tn antigen ((Tn Ag) or (GalNAca-Ser / Thr)), prostate- specific membrane antigen (PSMA), Receptor tyrosine kinase-like orphan receptor 1 (ROR1), Fms-Like Tyrosine Kinase 3 (FLT3), Tumor-associated glycoprotein 72 (TAG72), CD38, CD44v6, Carcinoembryonic antigen (CEA), Epithelial cell adhesion molecule (EPCAM), B7H3 (CD276), KIT (CD 117), Interleukin- 13 receptor subunit alpha-2, mesothelin, Interleukin 11 receptor alpha (IL-l lRa), prostate stem cell antigen (PSCA), Protease Serine 21, vascular endothelial growth factor receptor 2 (VEGFR2), Lewis(Y) antigen, CD24, Platelet-derived growth factor receptor beta (PDGFR-beta), Stage-specific embryonic antigen-4 (SSEA-4), CD20, Folate receptor alpha, HER2, HER3, Mucin 1, cell surface associated (MUC1), epidermal growth factor receptor (EGFR), neural cell adhesion molecule (NCAM), Prostase, prostatic acid phosphatase (PAP), elongation factor 2 mutated (ELF2M), Ephrin B2, fibroblast activation protein alpha (FAP), insulin-like growth factor 1 receptor (IGF-I receptor), carbonic anhydrase IX (CAIX), Proteasome (Prosome, Macropain) Subunit, Beta Type, 9 (LMP2), glycoprotein 100 (gplOO), oncogene fusion protein consisting of breakpoint cluster region (BCR) and Abelson murine leukemia viral oncogene homolog 1 (Abl) (bcr-abl), tyrosinase, ephrin type- A receptor 2 (EphA2), Fucosyl GM1, sialyl Lewis adhesion molecule (sLe), ganglioside GM3, transglutaminase 5 (TGS5), high molecular weight-melanoma-associated antigen (HMWMAA), o-acetyl-GD2 ganglioside (0AcGD2), Folate receptor beta, tumor endothelial marker 1 (TEM1 / CD248), tumor endothelial marker 7 -related (TEM7R), claudin 6 (CLDN6), claudin 18.2 (CLDN18.2), thyroid stimulating hormone receptor (TSHR), G protein-coupled receptor class C group 5, member D (GPRC5D), chromosome X open reading frame 61 (CXORF61), CD97, and CD179a. C. LNPs for Selective Delivery

[0308] In some embodiments, an LNP delivery vehicle delivers one or more encapsulated nucleic acid cargoes selectively to one or more specific cell populations of a subject. In some embodiments, an LNP delivery vehicle delivers one or more encapsulated nucleic acid cargoes selectively to one or more specific cell populations of a subject in vivo. In some embodiments, compositions of the present disclosure that induce activities of one or more polypeptides selectively within one or more specific cell populations of a subject, comprise an LNP delivery vehicle that selectively delivers one or more encapsulated nucleic acid cargoes to the one or more cell populations of the subject. In some embodiments, compositions of the present disclosure that induce activities of one or more polypeptides selectively within one or more cell populations of a subject in vivo, comprise an LNP delivery vehicle that selectively delivers one or more nucleic acid cargoes to the one or more cell populations of the subject in vivo. Without wishing to be bound by any particular theory, in some embodiments, an LNP delivery vehicle delivers one or more encapsulated nucleic acidcargoes encoding one or more polypeptides, each polypeptide having an activity, selectively to one or more specific cell populations of a subject, whereby at least one activity of the one or more polypeptides are then induced selectively in the one or more cell populations.

[0309] In some embodiments, an LNP delivery vehicle that delivers one or more encapsulated nucleic acid cargoes selectively to one or more specific cell populations of a subject is described as an LNP for selective delivery. In some embodiments, an LNP for selective delivery is an LNP for passive delivery (pLNP). In some embodiments, an LNP for selective delivery is an LNP for targeted delivery (tLNP). An LNP for passive delivery, as described in the instant specification, does not comprise a targeting moiety, as described herein, and delivers one or more encapsulated nucleic acid cargoes selectively to one or more specific cell populations of a subject.

[0310] In some embodiments, the pharmaceutical composition targets a tissue or an organ upon administration to a subject. In some embodiments, the tissue or the organ comprises the liver. In some embodiments, the tissue or the organ comprises a lymphoid organ. In some embodiments, the lymphoid organ comprises bone marrow, spleen, or lymph nodes. In some embodiments, the pharmaceutical composition targets a target cell upon administration to a subject. In some embodiments, the target cell comprises a hepatocyte, a lymphocyte, a leukocyte, a myeloid cell, or a hematopoietic stem cell. In some embodiments, the target cell comprises a B cell, and wherein the B cell comprises a plasmablast, a plasma cell, or a memory B cell. In some embodiments, the target cell comprises a T cell. In some embodiments, the target cell comprises an NK cell.

[0311] In some embodiments, the target cell comprises a cell in systemic circulation in the subject. ln some embodiments, the target cell comprises a cell within a tissue or an organ of the subject.

[0312] Alternatively, an LNP for targeted delivery, as described in the instant specification, comprises a targeting moiety, as described herein, and delivers one or more encapsulated nucleic acid cargoes selectively to one or more specific cell populations of a subject. In some embodiments, a pLNP, as described herein, may be modified with a targeting moiety to generate a corresponding tLNP. In some embodiments, a tLNP generated by modification of a pLNP with a targeting moiety may display more selective delivery relative to the initial pLNP without the targeting moiety. In some embodiments, a composition of the present disclosure comprises an LNP for passive delivery. In some embodiments, a composition of the present disclosure comprises an LNP for targeted delivery. In some embodiments, a composition of the present disclosure comprises both an LNP for passive delivery and an LNP for targeted delivery. i. LNPs For Targeted Delivery (tLNPs)

[0313] In some embodiments, a lipid nanoparticle for selective delivery, as described in the instant specification, is a lipid nanoparticle for targeted delivery (tLNP). As used in the instant specification, the term tLNP refers to an LNP comprising at least one targeting moiety that selectively binds a component of a membrane of a cell of a specific cell population, referred to herein as “thetargeting moiety’s target”. In some embodiments, the targeting moiety binds its target noncovalently. In some embodiments, the targeting moiety and the target form a covalent bond. In certain embodiments, a tLNP comprises more than one targeting moiety. In such embodiments, a tLNP may selectively deliver an encapsulated nucleic acid cargo to at least two specific cell populations.

[0314] In some embodiments, a targeting moiety is selected from the list consisting of: an antibody or antibody fragment, a protein, a glycan, a small molecule ligand, and an aptamer. In some embodiments, a targeting moiety is an antibody or antibody fragment. In some embodiments, a targeting moiety is an antibody. In some embodiments, a targeting moiety is an antibody fragment. In some embodiments, an antibody fragment is an antigen-binding fragment. In some embodiments, the antigen-binding fragment is selected from the group consisting of a Fab, a F(ab’)2 fragment, a scFv, a scab, a dAb, and a single domain antibody. In some embodiments, a targeting moiety that is an antibody or antibody fragment binds a target on cells of a specific cell population that is an antigen of the antibody or antibody fragment.

[0315] In some embodiments, a targeting moiety is a protein. In some embodiments, a targeting moiety that is a protein binds its target via a protein-protein interaction. In some embodiments, the protein-protein interaction mimics a native protein-protein interaction that occurs at the surface of cells of the specific cell population.

[0316] In some embodiments, a targeting moiety is a glycan. In some embodiments, a targeting moiety that is a glycan binds a target that is a lectin. In some embodiments, the glycan-lectin interaction mimics a native glycan-lectin interaction that occurs at the surface of cells of the specific cell population.

[0317] In some embodiments, a targeting moiety is a small molecule ligand. In some embodiments, a small molecule ligand is a native ligand for a receptor on the surface of a cell of a specific cell population. In some embodiments, the small molecule ligand-receptor interaction mimics a native ligand-receptor interaction that occurs at the surface of cells of the specific cell population. In some embodiments, a small molecule ligand is a drug.

[0318] In some embodiments, a targeting moiety is an aptamer.

[0319] In some embodiments, binding of a targeting moiety to its target triggers receptor mediated endocytosis (RME). Without wishing to be bound by any particular theory, in some embodiments, binding of a targeting moiety to its target may trigger RME, by which the tLNP and its contents are internalized into the cells of a specific cell population. In some embodiments, binding of a targeting moiety to its target may induce receptor signaling, potentially triggering cellular responses. In some embodiments, the cellular responses may help to enhance internalization of the tLNP and its contents into the cells of a specific cell population. In some embodiments, the cellular responses may enhance expression of the encapsulated nucleic acid cargo selectively delivered by the tLNP. In some embodiments, the cellular response may enhance proliferation of the cells of the specific cell population.

[0320] In some embodiments, the selectivity with which a tLNP delivers an encapsulated nucleic acid cargo to a specific cell population, or the potency of a polypeptide activity selectively induced in a specific cell population by a composition, can be enhanced by improving the affinity between a targeting moiety and its target. In some embodiments, improving the affinity between a targeting moiety and its target may reduce off-target effects and toxicity. In some embodiments, the affinity between a targeting moiety and its target can be measured in vitro. In some embodiments, the affinity between a targeting moiety and its target can be assessed by measuring a bimolecular dissociation constant (KD). In some embodiments, the KD between a targeting moiety and its target is at least about 500 nM, 400 nM, 300 nM, 200 nM, 100 nM, 50 nM, 10 nM, or 1 nM.

[0321] In some embodiments, an LNP for selective delivery is an LNP for targeted delivery. An LNP for targeted delivery, is an LNP for selective delivery as described herein, comprising at least one targeting moiety. In certain embodiments, the targeting moiety is conjugated to at least a portion of the PEG lipids of the LNP. In some embodiments, a targeting moiety is a moiety that recognizes or targets a molecular characteristic, as described herein, present on the surface of a cell. In some embodiments, recognition of a molecular characteristic of a cell by a targeting moiety facilitates contact of the LNP for targeted delivery comprising the targeting moiety with the cell comprising the molecular characteristic. Without wishing to be bound by any particular theory, in some embodiments, contact of a cell comprising a molecular characteristic with an LNP delivery vehicle comprising a targeting moiety that recognizes that molecular characteristic facilitates uptake of the LNP delivery vehicle by the cell. In some embodiments, compositions of the present disclosure comprise an LNP for targeted delivery, further comprising a targeting moiety that recognizes a molecular characteristic present on one or more cells of one or more specific cell populations. In some embodiments, an LNP for targeted delivery comprises one or more targeting moieties that recognize at least one molecular characteristic present on cells of a specific cell population. In some embodiments, an LNP for targeted delivery comprises one or more targeting moieties that recognize at least one molecular characteristic present on cells of at least two specific cell populations.

[0322] In some embodiments, a targeting moiety may be an antibody or a fragment thereof. In some embodiments, a molecular characteristic of a cell is an antigen. Accordingly, in some embodiments, a targeting moiety may be capable of binding to a target antigen. In certain embodiments, a LNP for targeted delivery comprises more than one targeting moiety. In certain embodiments, an LNP for targeted delivery comprises more than one targeting moiety, wherein each targeting moiety targets at least one distinct molecular characteristic. In some embodiments, the at least one distinct molecular characteristic is prevalent on different cells. In some embodiments, the different cells are part of the same specific cell population. In some embodiments, the different cells are part of different cell populations.

[0323] In some embodiments, a composition comprises a targeting moiety that is operably connected to an LNP delivery vehicle. In some embodiments, a targeting moiety is capable of bindingto a molecular characteristic that is a target antigen. In some embodiments, a target antigen is expressed by the cells of one or more specific cell populations. In some embodiments, one or more specific cell populations are present within a target organ. In some embodiments, a target antigen is expressed more by cells of one or more specific cell populations present in a target organ than it is by cell populations resident to the liver (i.e, hepatocytes).

[0324] In some embodiments, a targeting moiety is an antibody as described in WO2016189532A1, which is incorporated herein by reference. For example, in some embodiments, an LNP for targeted delivery comprises a targeting moiety that is a specific anti-CD38 monoclonal antibody (mAb), whereby an encapsulated nucleic acid cargo is selectively delivered to B-cell lymphocyte malignancies (such as MCL), without notable delivery toto other subtype leukocyte reference populations.

[0325] In some embodiments, a targeting moiety targets a molecular characteristic that is a cell surface receptor. In some embodiments, a targeting moiety targets a cell surface receptor selected from CCR7, CD3, CD4, CD5, CD8, CD16, CD19, CD20, CD21, CD22, CD25, CD28, CD35, CD40, CD45RA, CD45RO, CD52, CD62L, CD80, CD95, CD127, and CD137. In some embodiments, a targeting moiety targets a cell surface receptor selected from CD1, CD2, CD3, CD5, CD7, CD8, CD16, CD25, CD26, CD27, CD28, CD30, CD38, CD39, CD40L, CD44, CD45, CD62L, CD69, CD73, CD80, CD83, CD86, CD95, CD103, CD119, CD126, CD150, CD153, CD154, CD161, CD183, CD223, CD254, CD275, CD45RA, CXCR3, CXCR5, FasL, IL18R1, CTLA-4, 0X40, GITR, LAG3, ICOS, PD-1, leu-12, TCR, TLR1, TLR2, TLR3, TLR4, TLR6, NKG2D, CCR, CCR1, CCR2, CCR4, CCR6, and CCR7. In some embodiments, a targeting moiety targets a cell surface receptor selected from CD2, CD3, CD5 and CD7. In some embodiments, a targeting moiety targets a cell surface receptor selected from CD2, CD3, CD5, CD7, CD8, CD4, beta 7 integrin, beta 2 integrin, and C1q. In some embodiments, a targeting moiety targets CD117. In some embodiments, a targeting moiety targets a receptor selected from a mannose receptor, CD206 and C1q. In some embodiments, a targeting moiety is selected from T-cell receptor motif antibodies, T-cell α chain antibodies, T-cell β chain antibodies, T-cell γ chain antibodies, T-cell δ chain antibodies, CCR7 antibodies, CD3 antibodies, CD4 antibodies, CD5 antibodies, CD7 antibodies, CD8 antibodies, CD11b antibodies, CD11c antibodies, CD16 antibodies, CD19 antibodies, CD20 antibodies, CD21 antibodies, CD22 antibodies, CD25 antibodies, CD28 antibodies, CD34 antibodies, CD35 antibodies, CD40 antibodies, CD45RA antibodies, CD45RO antibodies, CD52 antibodies, CD56 antibodies, CD62L antibodies, CD68 antibodies, CD80 antibodies, CD95 antibodies, CD117 antibodies, CD127 antibodies, CD133 antibodies, CD137 (4-1BB) antibodies, CD163 antibodies, F4 / 80 antibodies, IL-4Rα antibodies, Sca-1 antibodies, CTLA-4 antibodies, GITR antibodies GARP antibodies, LAP antibodies, granzyme B antibodies, LFA-1 antibodies, transferrin receptor antibodies, and fragments thereof. In certain embodiments the targeting moiety is any one described or contemplated in US20230312713A1,US20230203538A1, US20230320995A1, US20160145348, and US20110038941, each of which is incorporated by reference herein in its entirety.

[0326] In some embodiments, an LNP for targeted delivery comprises a targeting moiety that recognizes a molecular characteristic that is a cell surface marker for a specific cell population. Without wishing to be bound by any particular theory, in some embodiments, a composition of the present disclosure comprises an LNP for targeted delivery further comprising a targeting moiety that recognizes a molecular characteristic that is a cell surface marker for a specific cell population, whereby one or more encapsulated nucleic acid cargoes encoding one or more polypeptides with at least one activity are selectively delivered to the specific cell population. For example, in some embodiments, an LNP for targeted delivery comprises a targeting moiety that recognizes a molecular characteristic that is a cell surface marker for T cell populations. Notable examples of cell surface markers for T cell populations, include but are not limited to CD2, CD3, CD4, CD5, CD7 and CD8. In some embodiments, an LNP for targeted delivery comprises a targeting moiety that recognizes a molecular characteristic that is a pan-T antigen. Notable examples of pan-T antigens include, but are not limited to CD2, CD3, CD5 and CD7. In some embodiments, an LNP for targeted delivery comprises a targeting moiety that is a cell surface marker for NK cell populations. ii. Delivery to At Least Two Cell Populations of a Subject

[0327] In some embodiments, an LNP for selective delivery selectively delivers a nucleic acid cargo to two or more specific cell populations of a subject. In these and other embodiments, an LNP for selective delivery independently delivers a nucleic acid cargo to a substantial fraction of each of the totals of the specific cell populations, while simultaneously delivering to a small fraction of the total of a reference cell population. The present disclosure teaches that when an LNP for selective delivery selectively delivers to two or more specific cell populations, that there is only one reference cell population. Accordingly, the present disclosure teaches that, in embodiments wherein compositions or methods of the present disclosure comprise an LNP for selective delivery that independently delivers a nucleic acid cargo to each of a first and second specific cell population, that the nucleic acid cargo is delivered to a substantial fraction of the total of the first specific cell population and a substantial fraction of the total of the second specific cell population, while simultaneously being delivered to a small fraction of the total of a single shared reference cell population. The present disclosure further teaches that, in certain embodiments, wherein compositions or methods of the present disclosure comprise an LNP for selective delivery that independently delivers a nucleic acid cargo to each of a first, second and third specific cell population, that the nucleic acid cargo is delivered to a substantial fraction of the total of the first specific cell population, a substantial fraction of the total of the second specific cell population and a substantial fraction of the total of the third specific cell population, while simultaneously being delivered to a small fraction of the total of a single shared reference cell population.

[0328] In some embodiments, an LNP of the present disclosure comprises an ionizable lipid, a structural lipid, a PEGylated lipid (aka PEG lipid), and a phospholipid. In alternative embodiments, an LNP comprises an ionizable lipid, a structural lipid, a PEGylated lipid (aka PEG lipid), and a zwitterionic amino acid lipid. In some embodiments, an LNP further comprises a 5th lipid, besides any of the aforementioned lipid components. In some embodiments, the LNP encapsulates one or more elements of the active agent of the present disclosure. In some embodiments, an LNP further comprises a targeting moiety covalently or non-covalently bound to the outer surface of the LNP. In some embodiments, the targeting moiety is a targeting moiety that binds to, or otherwise facilitates uptake by, cells of a particular organ system.

[0329] In certain embodiments, a tLNP comprises more than one targeting moiety, wherein the targeting moieties bind at least two different targets, and in some embodiments, the at least two different receptors are prevalent on different cell populations or tissues. In such embodiments, a tLNP may selectively deliver an encapsulated nucleic acid cargo to at least two specific cell populations. D. Methods of Measuring Selective Delivery

[0330] In some embodiments, compositions of the present disclosure comprise an LNP delivery vehicle and one or more encapsulated nucleic acid cargoes encoding one or more polypeptides with an activity. In some embodiments, an LNP delivery vehicle is an LNP for selective delivery. In some embodiments, an LNP for selective delivery, as described herein, is an LNP delivery vehicle that delivers one or more encapsulated nucleic acid cargoes to at least one specific cell population in an amount above a threshold value, while delivering the one or more nucleic acid cargoes to a reference cell population in an amount below a reference threshold. In some embodiments the threshold value and the reference threshold are the same. In some embodiments, the threshold value and the reference threshold are different. In some embodiments, the threshold value is greater than the reference threshold. In some embodiments, the reference threshold is greater than the threshold value. In some embodiments, a specific cell population is an immune cell population. In some embodiments, a reference cell population is a liver cell population (e.g., hepatocytes). For example, in exemplary embodiments, an LNP for selective delivery delivers one or more encapsulated nucleic acid cargoes to one or more immune cell populations in an amount above a threshold value, while delivering the one or more nucleic acid cargoes to liver cells at an amount below a reference threshold. In some embodiments, an immune cell population is a T cell or NK cell population. In some embodiments, one or more nucleic acid cargoes encode a chimeric antigen receptor (CAR) and / or chimeric stimulatory receptor (CSR).

[0331] In some embodiments, an LNP for selective delivery delivers at least two encapsulated nucleic acid cargoes to a specific cell population of a subject. In such embodiments, an LNP for selective delivery is so described if at least one encapsulated nucleic acid cargo is delivered to the specific cell population at an amount above the threshold value, while the same encapsulatednucleic acid cargo is delivered to the reference cell population at an amount below the reference threshold.

[0332] In some embodiments, an LNP for selective delivery delivers one or more encapsulated nucleic acid cargoes to at least two specific cell populations. For example, in some embodiments, an LNP for selective delivery delivers one or more encapsulated nucleic acid cargoes selectively to both a first and second specific cell population. In such embodiments, an LNP for selective delivery delivers one or more encapsulated nucleic acid cargoes to the first specific cell population at an amount above a first threshold value and delivers one or more encapsulated nucleic acid cargoes to the second specific cell population at an amount above a second threshold value.

[0333] In some embodiments, the first and second threshold values are each independently between about 5% and 95%. In some embodiments, the first and second threshold values are each about 5%. In some embodiments, the first and second threshold values are each about 10%. In some embodiments, the first and second threshold values are each about 15%. In some embodiments, the first and second threshold values are each about 20%. In some embodiments, the first and second threshold values are each about 25%. In some embodiments, the first and second threshold values are each about 30%. In some embodiments, the first and second threshold values are each about 35%. In some embodiments, the first and second threshold values are each about 40%. In some embodiments, the first and second threshold values are each about 45%. In some embodiments, the first and second threshold values are each about 50%. In some embodiments, the first and second threshold values are each about 55%. In some embodiments, the first and second threshold values are each about 60%. In some embodiments, the first and second threshold values are each about 65%. In some embodiments, the first and second threshold values are each about 70%. In some embodiments, the first and second threshold values are each about 75%. In some embodiments, the first and second threshold values are each about 80%. In some embodiments, the first and second threshold values are each about 85%. In some embodiments, the first and second threshold values are each about 90%. In some embodiments, the first and second threshold values are each about 95%. In certain embodiments, the first and second threshold values are not the same, such that one threshold value is higher than the other. In certain embodiments, the first threshold value is independently selected from 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 99%. In certain embodiments, the second threshold value is independently selected from 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 99%. In certain embodiments, the reference threshold value is independently selected from 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 99%.

[0334] In exemplary embodiments of the present disclosure, an LNP for selective delivery delivers a nucleic acid cargo to greater than 25% of the total of two or more cell populations, while simultaneously delivering to fewer than 5% of the total of a reference population. In some embodiments, an LNP for selective delivery delivers a nucleic acid cargo to greater than 25% of thetotal of a first cell population and greater than 25% of the total of a second cell population, while simultaneously delivering to fewer than 5% of the total of a reference population. In some embodiments, an LNP for selective delivery delivers a nucleic acid cargo to greater than 50% of the total of a first cell population and greater than 50% of the total of a second cell population, while simultaneously delivering to fewer than 20% of the total of a reference population. In some embodiments, an LNP for selective delivery delivers a nucleic acid cargo to greater than 60% of the total of a first cell population and greater than 60% of the total of a second cell population, while simultaneously delivering to fewer than 25% of the total of a reference population. In some embodiments, a reference cell population are hepatocytes. Accordingly, in some embodiments, an LNP for selective delivery of a nucleic acid cargo can be described as an LNP for extrahepatic delivery of a nucleic acid cargo.

[0335] In some embodiments, an LNP for selective delivery, as described herein, is an LNP delivery vehicle that delivers one or more encapsulated nucleic acid cargoes to at least one specific cell population in an amount greater than a reference LNP delivers to the same specific cell population under otherwise identical conditions. III. LNP payload

[0336] The instant specification describes compositions, methods, processes, kits and devices for the selection, design, preparation, manufacture, formulation, and / or use of LNP-based RNA medicines (e.g., vaccines, gene therapies, or gene-editing therapeutics). In various embodiments, the LNP-based RNA medicines comprise an LNP delivery system (as described in detail herein) and an encapsulated cargo / payload (e.g., RNA in the case of RNA medicines).

[0337] In the case of RNA medicines, the payload can be one or more RNA molecules, including coding RNA (e.g., linear or circular mRNA) or non-coding RNA (e.g., guide RNA, pegRNA, or retron ncRNA).

[0338] In various other embodiments, the payloads can include any type of nucleic acid molecule, including coding RNA molecules (e.g., mRNA), guide RNAs for editing systems (e.g., Cas9 guides, Cas12a guides, base editor guides, and prime editor guides), other non-coding RNAs relating to editing systems (e.g., retron ncRNAs), small RNAs (sRNAs)—which refer to a wide variety of polymeric RNA molecules that are generally less than 200 nucleotides in length with various functionalities, such as RNA interference, and include small-interfering RNA (siRNA), microRNAs (miRNA), piwi-interacting RNA (piRNA), repeat associated small interfering RNA (rasiRNA), small nuclear RNA (snRNA or U-RNA), small nucleolar RNA (snoRNA), small rDNA- derived RNA (srRNA), rRNA fragment (tRF), and Y RNA-derived small RNA, tRNA, rRNA, and self-amplifying RNA (saRNA)—and DNA molecules, such as DNA vectors, DNA plasmids, HDR donors, oligonucleotides, primers, etc., and chimeric molecules comprising both DNA and RNA. The cargo nucleic acid molecules may be single-stranded or double-stranded. Such nucleic acid cargomay comprise exactly one molecule. Such nucleic acid cargo may comprise exactly two molecules. Such nucleic acid cargo may comprise exactly three molecules. Such nucleic acid cargo may comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 distinct molecules. Such nucleic acid cargo may comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 distinct molecules. Such nucleic acid cargo may comprise between 1-25, or 5-30, or 10-35, or 20-40, or up to 100, or more distinct molecules.

[0339] In various other aspects, the LNPs described herein may be used to deliver any payload of interest to a biological target, e.g., to a cell or a bodily tissue. The term “payload” refers to an active substance (i.e., not limited to RNA or DNA), such as a small molecule, polypeptide, peptide, carbohydrate, or nucleic acid molecule, and includes, without limitation, mRNA molecules (including linear and circular mRNA) or non-coding RNA molecules (e.g., guide RNAs, pegRNAs, retron ncRNAs) which are encapsulated within the LNPs described herein. In some embodiments, the LNP cargo may comprise an RNP or ribonucleoprotein, such as a gene editing nuclease protein complexed with a cognate guide RNA.

[0340] In various embodiments, the payload is an RNA molecule, which may be linear or circular and may comprise one or more functional nucleotide sequences of interest, which may include, but are not limited to coding and non-coding nucleotide sequences. In various embodiments, the non-coding nucleotide sequences may comprise regulatory elements that influence RNA post- transcriptional processing, nuclear translation control sequences, and sequences which encode one or more biological products of interest, e.g., a therapeutic protein or antigen, among other sequence elements that may impact the functioning of the RNA or its encoded products. As used herein, the term “coding region of interest” or “product coding region” or the like may be used to refer to the encoded one or more biological products of interest. Equivalently, a product coding region may be referred to as a “product expression sequence.”

[0341] In various embodiments herein, the specification refers to “originator constructs” (or “originator polynucleotide constructs”) and “benchmark constructs” (or “benchmark polynucleotide constructs”), which are embodiments of payloads comprising nucleic acid molecules, i.e., embodiments of linear and / or circular mRNA payloads, and which may comprise a product coding region that encodes a polypeptide, such as, but not limited to an antigen or a therapeutic protein or to components of a gene editing system (e.g., a programmable nuclease).

[0342] FIG.2 shows an example of an originator construct 100, which may be a linear or circular mRNA molecule. The originator construct 100 may include at least one product coding region 10 which is or encodes a polypeptide of interest, such as, but not limited to a vaccine antigen or a therapeutic protein. The originator construct 100 may contain 1 or 2 flanking regions 20. The flanking regions 20 may be located 5' to the product coding region 10 and / or 3' to the product coding region 10. In some instances the originator construct 100 does not contain a flanking region 20. The flanking region 20 of the originator construct 100 may include at least one regulatory region 30. At least oneflanking region 20 of the originator polynucleotide construct 100 may include at least one identifier region 40. The identifier region 40 may be, but is not limited to, a barcode, label, signal and / or tag. Additionally, the identifier region 40 may be located within the product coding region 10 or may be located in the product coding region 10 and at least one flanking region 20.

[0343] In some embodiments, the originator construct comprises from about 5 to about 10,000 nucleotides in length. As a non-limiting examples, the length of the originator construct may be from 5 to 30, from 5 to 50, from 5 to 100, from 5 to 250, from 5 to 500, from 5 to 1,000, from 5 to 1,500, from 5 to 3,000 from 5 to 5,000, from 5 to 7,000, from 5 to 10,000 from 30 to 50, from 30 to 100, from 30 to 250, from 30 to 500, from 30 to 1,000, from 30 to 1,500, from 30 to 3,000, from 30 to 5,000, from 30 to 7,000, from 30 to 10,000, from 100 to 250, from 100 to 500, from 100 to 1,000, from 100 to 1,500, from 100 to 3,000, from 100 to 5,000, from 100 to 7,000, from 100 to 10,000, from 500 to 1,000, from 500 to 1,500, from 500 to 2,000, from 500 to 3,000, from 500 to 5,000, from 500 to 7,000, from 500 to 10,000, from 1,000 to 1,500, from 1,000 to 2,000, from 1,000 to 3,000, from 1,000 to 5,000, from 1,000 to 7,000, from 1,000 to 10,000, from 1,500 to 3,000, from 1,500 to 5,000, from 1,500 to 7,000, from 1,500 to 10,000, from 2,000 to 3,000, from 2,000 to 5,000, from 2,000 to 7,000, from 2,000 to 10,000, from 3,000 to 5,000, from 3,000 to 7,000, from 3,000 to 10,000, from 5,000 to 7,000, from 5,000 to 10,000, and from 7,000 to 10,000 nucleotides in length.

[0344] In some embodiments, the length of the product coding region is greater than about 5 nucleotides in length such as, but not limited to, at least or greater than about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 40, 45, 50, 55, 60, 70, 80, 90, 100, 120, 140, 160, 180, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1,000, 1,100, 1,200, 1,300, 1,400, 1,500, 1,600, 1,700, 1,800, 1,900, 2,000, 2,500, 3,000, 4,000, 5,000, 6,000, 7,000, 8,000, 9,000, 10,000 or more than 10,000 nucleotides in length.

[0345] In some embodiments, the flanking region may range independently from 0 to 10,000 nucleotides in length such as, but not limited to, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 40, 45, 50, 55, 60, 70, 80, 90, 100, 120, 140, 160, 180, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1,000, 1,100, 1,200, 1,300, 1,400, 1,500, 1,600, 1,700, 1,800, 1,900, 2,000, 2,500, 3,000, 4,000, 5,000, 6,000, 7,000, 8,000, 9,000, and 10,000 nucleotides in length.

[0346] In some embodiments, the regulatory region may range independently from 0 to 3,000 nucleotides in length such as, but not limited to, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 40, 45, 50, 55, 60, 70, 80, 90, 100, 120, 140, 160, 180, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1,000, 1,100, 1,200, 1,300, 1,400, 1,500, 1,600, 1,700, 1,800, 1,900, 2,000, 2,500, and 3,000 nucleotides in length.

[0347] In some embodiments, the originator construct may be circularized. In other embodiments, the originator construct may be concatemerized.

[0348] Originator constructs which include at least one identifier 40 or “identifier region” 40 (e.g., barcodes, labels, signals and / or tags) may also be referred to as “benchmark constructs” or “benchmark polynucleotide constructs.” The benchmark construct may comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more identifiers which may be the same or different throughout the benchmark polynucleotide construct.

[0349] In some embodiments, the identifier region may range independently from 1 to 3,000 nucleotides in length such as, but not limited to, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 40, 45, 50, 55, 60, 70, 80, 90, 100, 120, 140, 160, 180, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1,000, 1,100, 1,200, 1,300, 1,400, 1,500, 1,600, 1,700, 1,800, 1,900, 2,000, 2,500, and 3,000. As a non-limiting example the identifier region may be 1-5 residues, 2-5 residues, 3-5 residues, 2-7 residues, 3-7 residues, 1-10 residues, 2-10 residues, 3-10 residues, 5-10 residues, 7-10 residues, 1-15 residues, 2-15 residues, 3-15 residues, 5-15 residues, 7-15 residues, 10-15 residues, 12-15 residues, 1-20 residues, 2- 20 residues, 3-20 residues, 5-20 residues, 7-20 residues, 10-20 residues, 12-20 residues, 15-20 residues, 17-20 residues, 1-25 residues, 2-25 residues, 3-25 residues, 5-25 residues, 7-25 residues, 10- 25 residues, 12-25 residues, 15-25 residues, 17-25 residues, 20-25 residues, 1-30 residues, 2-30 residues, 3-30 residues, 5-30 residues, 7-30 residues, 10-30 residues, 12-30 residues, 15-30 residues, 17-30 residues, 20-30 residues, 25-30 residues, 1-35 residues, 2-35 residues, 3-35 residues, 5-35 residues, 7-35 residues, 10-35 residues, 12-35 residues, 15-35 residues, 17-35 residues, 20-35 residues, 25-35 residues, 30-35 residues, 1-35 residues, 2-35 residues, 3-35 residues, 5-35 residues, 7- 35 residues, 10-35 residues, 12-35 residues, 15-35 residues, 17-35 residues, 20-35 residues, 25-35 residues, 30-35 residues, 1-40 residues, 2-40 residues, 3-40 residues, 5-40 residues, 7-40 residues, 10- 40 residues, 12-40 residues, 15-40 residues, 17-40 residues, 20-40 residues, 25-40 residues, 30-40 residues, 35-40 residues, 1-45 residues, 2-45 residues, 3-45 residues, 5-45 residues, 7-45 residues, 10- 45 residues, 12-45 residues, 15-45 residues, 17-45 residues, 20-45 residues, 25-45 residues, 30-45 residues, 35-45 residues, 40-45 residues, 1-50 residues, 2-50 residues, 3-50 residues, 5-50 residues, 7- 50 residues, 10-50 residues, 12-50 residues, 15-50 residues, 17-50 residues, 20-50 residues, 25-50 residues, 30-50 residues, 35-50 residues, 40-50 residues, or 45-50 nucleotides in length.

[0350] In some embodiments, the identifier region in the benchmark construct overlaps with the product coding region. As used herein, "overlap" means that at least one nucleotide of the identifier region extends into the product coding region. In some aspects the identifier region overlaps with the product coding region by 1 nucleotide, 2 nucleotides, 3 nucleotides, 4 nucleotides, 5 nucleotides, 6 nucleotides, 7 nucleotides, 8 nucleotides, 9 nucleotides, 10 nucleotides, 11 nucleotides, 12 nucleotides, 13 nucleotides, 14 nucleotides, 15 nucleotides, 16 nucleotides, 17 nucleotides, 18 nucleotides, 19 nucleotides, 20 nucleotides, 21 nucleotides, 22 nucleotides, 23 nucleotides, 24 nucleotides, 25 nucleotides, 26 nucleotides, 27 nucleotides, 28 nucleotides, 29 nucleotides, 30 nucleotides, 31 nucleotides, 32 nucleotides, 33 nucleotides, 34 nucleotides, 35 nucleotides, 36nucleotides, 37 nucleotides, 38 nucleotides, 39 nucleotides, 40 nucleotides 41 nucleotides, 42 nucleotides, 43 nucleotides, 44 nucleotides, 45 nucleotides, 46 nucleotides, 47 nucleotides, 48 nucleotides, 49 nucleotides, 50 nucleotides or more than 50 nucleotides. In some aspects the identifier region overlaps with the product coding region by 1-5 nucleotides, 2-5 nucleotides, 3-5 nucleotides, 2- 7 nucleotides, 3-7 nucleotides, 1-10 nucleotides, 2-10 nucleotides, 3-10 nucleotides, 5-10 nucleotides, 7-10 nucleotides, 1-15 nucleotides, 2-15 nucleotides, 3-15 nucleotides, 5-15 nucleotides, 7-15 nucleotides, 10-15 nucleotides, 12-15 nucleotides, 1-20 nucleotides, 2-20 nucleotides, 3-20 nucleotides, 5-20 nucleotides, 7-20 nucleotides, 10-20 nucleotides, 12-20 nucleotides, 15-20 nucleotides, 17-20 nucleotides, 1-25 nucleotides, 2-25 nucleotides, 3-25 nucleotides, 5-25 nucleotides, 7-25 nucleotides, 10-25 nucleotides, 12-25 nucleotides, 15-25 nucleotides, 17-25 nucleotides, 20-25 nucleotides, 1-30 nucleotides, 2-30 nucleotides, 3-30 nucleotides, 5-30 nucleotides, 7-30 nucleotides, 10-30 nucleotides, 12-30 nucleotides, 15-30 nucleotides, 17-30 nucleotides, 20-30 nucleotides, 25-30 nucleotides, 1-35 nucleotides, 2-35 nucleotides, 3-35 nucleotides, 5-35 nucleotides, 7-35 nucleotides, 10-35 nucleotides, 12-35 nucleotides, 15-35 nucleotides, 17-35 nucleotides, 20-35 nucleotides, 25-35 nucleotides, 30-35 nucleotides, 1-35 nucleotides, 2-35 nucleotides, 3-35 nucleotides, 5-35 nucleotides, 7-35 nucleotides, 10-35 nucleotides, 12-35 nucleotides, 15-35 nucleotides, 17-35 nucleotides, 20-35 nucleotides, 25-35 nucleotides, 30-35 nucleotides, 1-40 nucleotides, 2-40 nucleotides, 3-40 nucleotides, 5-40 nucleotides, 7-40 nucleotides, 10-40 nucleotides, 12-40 nucleotides, 15-40 nucleotides, 17-40 nucleotides, 20-40 nucleotides, 25-40 nucleotides, 30-40 nucleotides, 35-40 nucleotides, 1-45 nucleotides, 2-45 nucleotides, 3-45 nucleotides, 5-45 nucleotides, 7-45 nucleotides, 10-45 nucleotides, 12-45 nucleotides, 15-45 nucleotides, 17-45 nucleotides, 20-45 nucleotides, 25-45 nucleotides, 30-45 nucleotides, 35-45 nucleotides, 40-45 nucleotides, 1-50 nucleotides, 2-50 nucleotides, 3-50 nucleotides, 5-50 nucleotides, 7-50 nucleotides, 10-50 nucleotides, 12-50 nucleotides, 15-50 nucleotides, 17-50 nucleotides, 20-50 nucleotides, 25-50 nucleotides, 30-50 nucleotides, 35-50 nucleotides, 40-50 nucleotides, or 45-50 nucleotides.

[0351] In some embodiments, the benchmark polynucleotide construct comprises a product coding region and an identifier region. The identifier region may be located 5' to the product coding region, 3' to the product coding region, or the identifier region may overlap with the 5' end or the 3'end of the product coding region.

[0352] In some embodiments, the benchmark polynucleotide construct comprises a product coding region and two identifier regions. Each identifier region may independently be located 5' to the product coding region, 3' to the product coding region, or the identifier region may overlap with the 5' end or the 3'end of the product coding region.

[0353] As a non-limiting example, the first identifier region is located 5' to the product coding region and the second identifier region is located 3' to the product coding region. As a non- limiting example, the first and second identifier regions are located 5' to the product coding region. Asa non-limiting example, the first and second identifier regions are located 3' to the product coding region.

[0354] As a non-limiting example, the first identifier region is inverted and is located 5' to the product coding region and the second identifier region is located 3' to the product coding region. As a non-limiting example, the first identifier region is inverted and is located 5' to the product coding region and the second identifier region is inverted and is located 3' to the product coding region. As a non-limiting example, the first identifier region is located 5' to the product coding region and the second identifier region is inverted and is located 3' to the product coding region. As a non-limiting example, the first and second identifier regions are both inverted and are located 5' to the product coding region. As a non-limiting example, the first and second identifier regions are located 5' to the product coding region and the first identifier region is inverted. As a non-limiting example, the first and second identifier regions are located 5' to the product coding region and the second identifier region is inverted. As a non-limiting example, the first and second identifier region are both inverted and located 3' to the product coding region. As a non-limiting example, the first and second identifier regions are located 3' to the product coding region and the first identifier region is inverted. As a non- limiting example, the first and second identifier regions are located 3' to the product coding region and the second identifier region is inverted.

[0355] As a non-limiting example, the first identifier region is located 5' to the product coding region and overlaps with the product coding region and the second identifier region is located 3' to the product coding region. As a non-limiting example, the first identifier region is located 5' to the product coding region and the second identifier region is located 3' to the product coding region and overlaps with the product coding region.

[0356] As a non-limiting example, the first and second identifier regions are located 5' to the product coding region and the second identifier region overlaps with the product coding region. As a non-limiting example, the first and second identifier regions are located 3' to the product coding region and the first identifier region overlaps with the product coding region.

[0357] As a non-limiting example, the first identifier region is inverted, is located 5' to the product coding region and overlaps with the product coding region, and the second identifier region is located 3' to the product coding region. As a non-limiting example, the first identifier region is inverted and is located 5' to the product coding region and the second identifier region is located 3' to the product coding region and overlaps with the product coding region. As a non-limiting example, the first identifier region is inverted, is located 5' to the product coding region, the second identifier region is located 3' to the product coding region, and both of the first and second identifier regions overlap with the product coding region.

[0358] As a non-limiting example, the first identifier region is inverted, is located 5' to the product coding region and overlaps with the product coding region, and the second identifier region is inverted and is located 3' to the product coding region. As a non-limiting example, the first identifierregion is inverted and is located 5' to the product coding region and the second identifier region is inverted, is located 3' to the product coding region and overlaps with the product coding region. As a non-limiting example, the first identifier region is inverted and is located 5' to the product coding region, and the second identifier region is inverted and is located 3' to the product coding region, and both of the first and second identifier regions overlap with the product coding region.

[0359] As a non-limiting example, the first identifier region is located 5' to the product coding region and overlaps with the product coding region, and the second identifier region is inverted and is located 3' to the product coding region. As a non-limiting example, the first identifier region is located 5' to the product coding region and the second identifier region is inverted, is located 3' to the product coding region and overlaps with the product coding region. As a non-limiting example, the first identifier region is located 5' to the product coding region and the second identifier region is inverted and is located 3' to the product coding region, and both of the first and second identifier regions overlap with the product coding region.

[0360] As a non-limiting example, the first and second identifier regions are both inverted and are located 5' to the product coding region, and the second identifier region overlaps with the product coding region. As a non-limiting example, the first and second identifier regions are located 5' to the product coding region and the first identifier region is inverted, and the second identifier region overlaps with the product coding region. As a non-limiting example, the first and second identifier regions are located 5' to the product coding region and the second identifier region is inverted and overlaps with the product coding region. As a non-limiting example, the first and second identifier region are both inverted and located 3' to the product coding region, and the first identifier region overlap with the product coding region. As a non-limiting example, the first and second identifier regions are located 3' to the product coding region and the first identifier region is inverted and overlaps with the product coding region. As a non-limiting example, the first and second identifier regions are located 3' to the product coding region and the second identifier region is inverted, and the first product coding region overlap with the product coding region.

[0361] In some embodiments, at least one identifier moiety may be associated with the benchmark polynucleotide construct. The benchmark polynucleotide construct may have 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more identifier moieties associated with the benchmark polynucleotide construct which may be the same moiety or different moieties associated with the benchmark polynucleotide construct. Each identifier moiety may independently be located on the flanking region 5' to the product coding region, on the flanking region 3' to the product coding region, or the location of the identifier moiety may span the 5' end or the 3'end of the product coding region and a flanking region. In some aspects the location of the identifier moiety may include one or more nucleotides of the product coding region such as, but not limited to, 1 nucleotide, 2 nucleotides, 3 nucleotides, 4 nucleotides, 5 nucleotides, 6 nucleotides, 7 nucleotides, 8 nucleotides, 9 nucleotides, 10 nucleotides, 11 nucleotides, 12 nucleotides, 13 nucleotides, 14 nucleotides, 15 nucleotides, 16 nucleotides, 17nucleotides, 18 nucleotides, 19 nucleotides, 20 nucleotides, 21 nucleotides, 22 nucleotides, 23 nucleotides, 24 nucleotides, 25 nucleotides, 26 nucleotides, 27 nucleotides, 28 nucleotides, 29 nucleotides, 30 nucleotides, 31 nucleotides, 32 nucleotides, 33 nucleotides, 34 nucleotides, 35 nucleotides, 36 nucleotides, 37 nucleotides, 38 nucleotides, 39 nucleotides, 40 nucleotides 41 nucleotides, 42 nucleotides, 43 nucleotides, 44 nucleotides, 45 nucleotides, 46 nucleotides, 47 nucleotides, 48 nucleotides, 49 nucleotides, 50 nucleotides or more than 50 nucleotides. In some aspects the location of the identifier moiety may include one or more nucleotides of the product coding region such as, but not limited to, 1-5 nucleotides, 2-5 nucleotides, 3-5 nucleotides, 2-7 nucleotides, 3-7 nucleotides, 1-10 nucleotides, 2-10 nucleotides, 3-10 nucleotides, 5-10 nucleotides, 7-10 nucleotides, 1-15 nucleotides, 2-15 nucleotides, 3-15 nucleotides, 5-15 nucleotides, 7-15 nucleotides, 10-15 nucleotides, 12-15 nucleotides, 1-20 nucleotides, 2-20 nucleotides, 3-20 nucleotides, 5-20 nucleotides, 7-20 nucleotides, 10-20 nucleotides, 12-20 nucleotides, 15-20 nucleotides, 17-20 nucleotides, 1-25 nucleotides, 2-25 nucleotides, 3-25 nucleotides, 5-25 nucleotides, 7-25 nucleotides, 10-25 nucleotides, 12-25 nucleotides, 15-25 nucleotides, 17-25 nucleotides, 20-25 nucleotides, 1-30 nucleotides, 2-30 nucleotides, 3-30 nucleotides, 5-30 nucleotides, 7-30 nucleotides, 10-30 nucleotides, 12-30 nucleotides, 15-30 nucleotides, 17-30 nucleotides, 20-30 nucleotides, 25-30 nucleotides, 1-35 nucleotides, 2-35 nucleotides, 3-35 nucleotides, 5-35 nucleotides, 7-35 nucleotides, 10-35 nucleotides, 12-35 nucleotides, 15-35 nucleotides, 17-35 nucleotides, 20-35 nucleotides, 25-35 nucleotides, 30-35 nucleotides, 1-35 nucleotides, 2-35 nucleotides, 3-35 nucleotides, 5-35 nucleotides, 7-35 nucleotides, 10-35 nucleotides, 12-35 nucleotides, 15-35 nucleotides, 17-35 nucleotides, 20-35 nucleotides, 25-35 nucleotides, 30-35 nucleotides, 1-40 nucleotides, 2-40 nucleotides, 3-40 nucleotides, 5-40 nucleotides, 7-40 nucleotides, 10-40 nucleotides, 12-40 nucleotides, 15-40 nucleotides, 17-40 nucleotides, 20-40 nucleotides, 25-40 nucleotides, 30-40 nucleotides, 35-40 nucleotides, 1-45 nucleotides, 2-45 nucleotides, 3-45 nucleotides, 5-45 nucleotides, 7-45 nucleotides, 10-45 nucleotides, 12-45 nucleotides, 15-45 nucleotides, 17-45 nucleotides, 20-45 nucleotides, 25-45 nucleotides, 30-45 nucleotides, 35-45 nucleotides, 40-45 nucleotides, 1-50 nucleotides, 2-50 nucleotides, 3-50 nucleotides, 5-50 nucleotides, 7-50 nucleotides, 10-50 nucleotides, 12-50 nucleotides, 15-50 nucleotides, 17-50 nucleotides, 20-50 nucleotides, 25-50 nucleotides, 30-50 nucleotides, 35-50 nucleotides, 40-50 nucleotides, or 45-50 nucleotides.

[0362] In some embodiments, one identifier moiety may be associated with the benchmark polynucleotide construct. As a non-limiting example, the identifier moiety may be associated with the benchmark polynucleotide construct on the 5' end of the benchmark polynucleotide construct. As a non-limiting example, the identifier moiety may be associated with the benchmark polynucleotide construct on the 5' flanking region. As a non-limiting example, the identifier moiety may be associated with the benchmark polynucleotide construct on the 3' flanking region. As a non-limiting example, the identifier moiety may be associated with the benchmark polynucleotide construct on the 3' end of the benchmark polynucleotide construct. As a non-limiting example, the identifier moietymay be associated with the benchmark polynucleotide construct on the product coding region. As a non-limiting example, the benchmark polynucleotide construct comprises an identifier moiety and the location of the identifier moiety spans the 5' end of the product coding region and the 5' flanking region. As a non-limiting example, the benchmark polynucleotide construct comprises an identifier moiety and the location of the identifier moiety spans the 3' end of the product coding region and the 3' flanking region.

[0363] In some embodiments, two identifier moieties are associated with the benchmark polynucleotide construct. As a non-limiting example, the first identifier moiety and the second identifier moiety are located on the 5' flanking region. As a non-limiting example, the first identifier moiety and the second identifier moiety are located on the product coding region. As a non-limiting example, the first identifier moiety and the second identifier moiety are located on the 3' flanking region. As a non-limiting example, the first identifier moiety and the second identifier moiety are located on the 5' end of the benchmark polynucleotide construct. As a non-limiting example, the first identifier moiety and the second identifier moiety are located on the 3' end of the benchmark polynucleotide construct.

[0364] As a non-limiting example, the first identifier moiety is located on the 5' end of the benchmark polynucleotide construct and the second identifier moiety is located on the 5' flanking region. As a non-limiting example, the first identifier moiety is located on the 5' end of the benchmark polynucleotide construct and the second identifier moiety is located on the product coding region. As a non-limiting example, the first identifier moiety is located on the 5' end of the benchmark polynucleotide construct and the second identifier moiety is located on the 3' flanking region. As a non-limiting example, the first identifier moiety is located on the 5' end of the benchmark polynucleotide construct and the location of the second identifier moiety spans the 5' flanking region and the product coding region. As a non-limiting example, the first identifier moiety is located on the 5' end of the benchmark polynucleotide construct and the location of the second identifier moiety spans the 3' flanking region and the product coding region. As a non-limiting example, the first identifier moiety is located on the 5' end of the benchmark polynucleotide construct and the second identifier moiety is located on the 3' end of the benchmark polynucleotide construct.

[0365] As a non-limiting example, the first identifier moiety is located on the 5' flanking region and the second identifier moiety is located on the product coding region. As a non-limiting example, the first identifier moiety is located on the 5' flanking region and the second identifier moiety is located on the 3' flanking region. As a non-limiting example, the first identifier moiety is located on the 5' flanking region and the location of the second identifier moiety spans the 5' flanking region and the product coding region. As a non-limiting example, the first identifier moiety is located on the 5' flanking region and the location of the second identifier moiety spans the 3' flanking region and the product coding region. As a non-limiting example, the first identifier moiety is located on the 5' flanking region and the second identifier moiety is located on the 5' end of the benchmarkpolynucleotide construct. As a non-limiting example, the first identifier moiety is located on the 5' flanking region and the second identifier moiety is located on the 3' end of the benchmark polynucleotide construct.

[0366] As a non-limiting example, the location of the first identifier moiety spans the 5' flanking region and the product coding region and the second identifier moiety is located on the 5' end of the benchmark polynucleotide construct. As a non-limiting example, the location of the first identifier moiety spans the 5' flanking region and the product coding region and the second identifier moiety is located on the 5' flanking region. As a non-limiting example, the location of the first identifier moiety spans the 5' flanking region and the product coding region and the second identifier moiety is located on the product coding region. As a non-limiting example, the location of the first identifier moiety spans the 5' flanking region and the product coding region and the location of the second identifier moiety spans the 3' flanking region and the product coding region. As a non-limiting example, the location of the first identifier moiety spans the 5' flanking region and the product coding region and the second identifier moiety is located on the 3' flanking region. As a non-limiting example, the location of the first identifier moiety spans the 5' flanking region and the product coding region and the second identifier moiety is located on the 3' end of the benchmark polynucleotide construct.

[0367] As a non-limiting example, the first identifier moiety is located on the product coding region and the second identifier moiety is located on the 5' end of the benchmark polynucleotide construct. As a non-limiting example, the first identifier moiety is located on the product coding region and the second identifier moiety is located on the 5' flanking region. As a non-limiting example, the first identifier moiety is located on the product coding region and the location of the second identifier moiety spans the 5' flanking region and the product coding region. As a non-limiting example, the first identifier moiety is located on the product coding region and the location of the second identifier moiety spans the 3' flanking region and the product coding region. As a non-limiting example, the first identifier moiety is located on the product coding region and the second identifier moiety is located on the 3' flanking region. As a non-limiting example, the first identifier moiety is located on the product coding region and the second identifier moiety is located on the 3' end of the benchmark polynucleotide construct.

[0368] As a non-limiting example, the location of the first identifier moiety spans the 3' flanking region and the product coding region and the second identifier moiety is located on the 5' end of the benchmark polynucleotide construct. As a non-limiting example, the location of the first identifier moiety spans the 3' flanking region and the product coding region and the second identifier moiety is located on the 5' flanking region. As a non-limiting example, the location of the first identifier moiety spans the 3' flanking region and the product coding region and the location of the second identifier moiety spans the 5' flanking region and the product coding region. As a non-limiting example, the location of the first identifier moiety spans the 3' flanking region and the product codingregion and the second identifier moiety is located on the product coding region. As a non-limiting example, the location of the first identifier moiety spans the 3' flanking region and the product coding region and the second identifier moiety is located on the 3' flanking region. As a non-limiting example, the location of the first identifier moiety spans the 3' flanking region and the product coding region and the second identifier moiety is located on the 3'end of the benchmark polynucleotide construct.

[0369] As a non-limiting example, the location of the first identifier moiety spans the 3' flanking region and the product coding region and the second identifier moiety is located on the 5' flanking region. As a non-limiting example, the location of the first identifier moiety spans the 5' flanking region and the product coding region and the second identifier moiety is located on the product coding region. As a non-limiting example, the location of the first identifier moiety spans the 5' flanking region and the product coding region and the location of the second identifier moiety spans the 3' flanking region and the product coding region. As a non-limiting example, the location of the first identifier moiety spans the 5' flanking region and the product coding region and the second identifier moiety is located on the 3' flanking region. As a non-limiting example, the location of the first identifier moiety spans the 5' flanking region and the product coding region and the second identifier moiety is located on the 3' end of the benchmark polynucleotide construct.

[0370] As a non-limiting example, the first identifier moiety is located on the 3' flanking region and the second identifier moiety is located on the 5' end of the benchmark polynucleotide construct. As a non-limiting example, the first identifier moiety is located on the 3' flanking region and the second identifier moiety is located on the 5' flanking region. As a non-limiting example, the first identifier moiety is located on the 3' flanking region and the location of the second identifier moiety spans the 5' flanking region and the product coding region. As a non-limiting example, the first identifier moiety is located on the 3' flanking region and the second identifier moiety is located on the product coding region. As a non-limiting example, the first identifier moiety is located on the 3' flanking region and the location of the second identifier moiety spans the 3' flanking region and the product coding region. As a non-limiting example, the first identifier moiety is located on the 3' flanking region and the second identifier moiety is located on the 3' end of the benchmark polynucleotide construct.

[0371] As a non-limiting example, the first identifier moiety is located on the 3' end of the benchmark polynucleotide construct and the second identifier moiety is located on the 5' end of the benchmark polynucleotide construct. As a non-limiting example, the first identifier moiety is located on the 3' end of the benchmark polynucleotide construct and the second identifier moiety is located on the 5' flanking region. As a non-limiting example, the first identifier moiety is located on the 5' end of the benchmark polynucleotide construct and the location of the second identifier moiety spans the 5' flanking region and the product coding region. As a non-limiting example, the first identifier moiety is located on the 3' end of the benchmark polynucleotide construct and the second identifier moiety islocated on the product coding region. As a non-limiting example, the first identifier moiety is located on the 5' end of the benchmark polynucleotide construct and the location of the second identifier moiety spans the 3' flanking region and the product coding region. As a non-limiting example, the first identifier moiety is located on the 3' end of the benchmark polynucleotide construct and the second identifier moiety is located on the 3' flanking region.

[0372] In some embodiments, three identifier moieties are associated with the benchmark polynucleotide construct. In some embodiments, four identifier moieties are associated with the benchmark polynucleotide construct. In some embodiments, five identifier moieties are associated with the benchmark polynucleotide construct. In some embodiments, six identifier moieties are associated with the benchmark polynucleotide construct. In some embodiments, seven identifier moieties are associated with the benchmark polynucleotide construct. In some embodiments, eight identifier moieties are associated with the benchmark polynucleotide construct. In some embodiments, nine identifier moieties are associated with the benchmark polynucleotide construct. In some embodiments, ten identifier moieties are associated with the benchmark polynucleotide construct.

[0373] In some embodiments, the product coding region encodes a biologically active molecule such as, but not limited to a therapeutic protein or an antigen. As used herein, the term "biologically active" refers to a characteristic of any agent that has activity in a biological system, and particularly in an organism. For instance, an agent that, when administered to an organism, has a biological effect on that organism, is considered to be biologically active. In some embodiments, the CROI encodes one or more prophylactically- or therapeutically-active proteins, polypeptides, or other factors. As a non-limiting example, the CROI may encode an agent that enhances tumor killing activity such as, but not limited to, TRAIL or tumor necrosis factor (TNF), in a cancer. As another non-limiting example, the CROI may encode an agent suitable for the treatment of conditions such as muscular dystrophy (e.g., CROI encodes Dystrophin), cardiovascular disease (e.g., CROI encodes SERCA2a, GATA4, Tbx5, Mef2C, Hand2, Myocd, etc.), neurodegenerative disease (e.g., CROI encodes NGF, BDNF, GDNF, NT-3, etc.), chronic pain (e.g., CROI encodes GlyRal), an enkephalin, or a glutamate decarboxylase (e.g., CROI encodes GAD65, GAD67, or another isoform), lung disease (e.g., CROI encodes CFTR), hemophilia (e.g., CROI encodes Factor VIII or Factor IX), neoplasia (e.g., CROI encodes PTEN, ATM, ATR, EGFR, ERBB2, ERBB3, ERBB4, Notchl, Notch2, Notch3, Notch4, AKT, AKT2, AKT3, HIF, HI Fla, HIF3a, Met, HRG, Bcl2, PPARalpha, PPAR gamma, WT1 (Wilms Tumor), FGF Receptor Family members (5 members: 1, 2, 3, 4, 5), CDKN2a, APC, RB (retinoblastoma), MEN1, VHL, BRCA1, BRCA2, AR (Androgen Receptor), TSG101, IGF, IGF Receptor, Igfl (4 variants), Igf2 (3 variants), Igfl Receptor, Igf2 Receptor, Bax, Bcl2, caspases family (9 members: 1, 2, 3, 4, 6, 7, 8, 9, 12), Kras, Ape), age-related macular degeneration (e.g., CROI encodes Aber, Ccl2, Cc2, cp (ceruloplasmin), Timp3, cathepsin D, Vldlr), schizophrenia (e.g. Neuregulin (Nrgl), Erb4 (receptor for Neuregulin), Complexin-l (Cplxl), Tphl Tryptophan hydroxylase, Tph2 Tryptophan hydroxylase 2, Neurexin 1, GSK3, GSK3a, GSK3b, 5-HIT (Slc6a4),COMT, DRD (Drdla), SLC6A3, DAOA, DTNBPI, Dao (Daol)), trinucleotide repeat disorders (e.g., HTT (Huntington's Dx), SBMA / SMAXI / AR (Kennedy's Dx), FXN / X25 (Friedrich's Ataxia), ATX3 (Machado-Joseph's Dx), ATXNI and ATXN2 (spinocerebellar ataxias), DMPK (myotonic dystrophy), Atrophin-1 and Atnl(DRPLA Dx), CBP (Creb-BP-global instability), VLDLR (Alzheimer's), Atxn7, Atxn10), fragile X syndrome (e.g., CROI encodes FMR2, FXRI, FXR2, mGLUR5), secretase related disorders (e.g., CROI encodes APH-1 (alpha and beta), Presenilin (Psenl), nicastrin (Ncstn), PEN-2), ALS (e.g., CROI encodes SOD1, ALS2, STEX, FUS, TARD BP, VEGF (VEGF-a, VEGF-b, VEGF- c)), autism (e.g., CROI encodes Mecp2, BZRAP1, MDGA2, Sema5A, Neurexin 1), Alzheimer's disease (e.g., CROI encodes El, CHIP, UCH, UBB, Tau, LRP, PICALM, Clusterin, PS1, SORL1, CR1, Vldlr, Ubal, Uba3, CHIP28 (Aqpl, Aquaporin 1), Uchll, Uchl3, APP), inflammation (e.g., CROI encodes IL-10, IL-1 (IL-Ia, IL-Ib), IL-13, IL-17 (IL-17a (CTLA8), IL-17b, IL-17c, IL-17d, IL-171), 11-23, Cx3crl, ptpn22, TNFa, NOD2 / CARD15 for IBD, IL-6, IL-12 (IL-12a, IL-12b), CTLA4, Cx3cll), Parkinson's Disease (e.g., x-Synuclein, DJ-1, LRRK2, Parkin, PINK1), blood and coagulation disorders, such as, e.g., anemia, bare lymphocyte syndrome, bleeding disorders, hemophagocytic lymphohistiocytosis disorders, hemophilia A, hemophilia B, hemorrhagic disorders, leukocyte deficiencies and disorders, sickle cell anemia, and thalassemia (e.g., CROI encodes CRAN1, CDA1, RPS19, DBA, PKLR, PK1, NT5C3, UMPH1, PSNI, RHAG, RH50A, NRAMP2, SPTB, ALAS2, ANH1, ASB, ABCB7, ABC7, ASAT, TAPBP, TPSN, TAP2, ABCB3, PSF2, RING11, MHC2TA, C2TA, RFX5, RFXAP, RFX5, TBXA2R, P2RX1, P2X1, HF1, CFH, HUS, MCFD2, FANCA, FAC A, FA1, FA, FA A, FAAP95, FAAP90, FLJ34064, FANCB, FANCC, FACC, BRCA2, FANCDI, FANCD2, FANCD, FACD, FAD, FANCE, FACE, FANCF, XRCC9, FANCG, BR1PI, BACH1, FANCJ, PHF9, FANCL, FANCM, KIAA1596, PRF1, HPLH2, UNC13D, MUNC13-4, HPLH3, HLH3, FHL3, F8, FSC, PI, ATT, F5, ITGB2, CD18, LCAMB, LAD, EIF2B1, EIF2BA, EIF2B2, EIF2B3, EIF2B5, LVWM, CACH, CLE, EIF2B4, HBB, HBA2, HBB, HBD, LCRB, HBA1), B-cell non-Hodgkin lymphoma or leukemia (e.g., CROI encodes BCL7A, BCL7, ALI, TCL5, SCL, TAL2, FLT3, NBS1, NBS, ZNFN1AI, 1KI, LYF1, HOXD4, HOX4B, BCR, CML, PHL, ALL, ARNT, KRAS2, RASK2, GMPS, AFIO, ARHGEF12, LARG, KIAA0382, CALM, CLTH, CEBPA, CEBP, CHIC2, BTL, FLT3, KIT, PBT, LPP, NPMI, NUP214, D9S46E, CAN, CAIN, RUNXI, CBFA2, AML1, WHSC1LI, NSD3, FLT3, AF1Q, NPMI, NUMA1, ZNF145, PLZF, PML, MYL, STAT5B, AF1Q, CALM, CLTH, ARL11, ARLTS1, P2RX7, P2X7, BCR, CML, PHL, ALL, GRAF, NF1, VRNF, WSS, NFNS, PTPNII, PTP2C, SHP2, NS1, BCL2, CCND1, PRAD1, BCL1, TCRA, GATA1, GF1, ERYF1, NFE1, ABLI, NQO1, DIA4, NMOR1, NUP214, D9S46E, CAN, CAIN), inflammation and immune related diseases and disorders (e.g., CROI encodes KIR3DL1, NKAT3, NKB1, AMB11, K1R3DS1, IFNG, CXCL12, TNFRSF6, APT1, FAS, CD95, ALPS1A, IL2RG, SCIDX1, SCIDX, IMD4, CCL5, SCYA5, D17S136E, TCP228, IL10, CSIF, CMKBR2, CCR2, CMKBR5, CCCKR5 (CCR5), CD3E, CD3G, AICDA, AID, HIGM2, TNFRSF5, CD40, UNG, DGU, HIGM4, TNFSFS, CD40LG, HIGM1, IGM, FOXP3, IPEX, AIID, XPID, PIDX,TNFRSF14B, TACI), inflammation (e.g., CROI encodes IL-10, IL-1 (IL-IA, IL-IB), IL-13, IL-17 (IL- 17a (CTLA8), IL-17b, IL-17c, IL-17d, IL-171), 11-23, Cx3crl, ptpn22, TNFa, NOD2 / CARD15 for IBD, IL-6, IL-12 (IL-12a, IL-12b), CTLA4, Cx3cII), JAK3, JAKL, DCLREIC, ARTEMIS, SCIDA, RAG1, RAG2, ADA, PTPRC, CD45, LCA, IL7R, CD3D, T3D, IL2RG, SCIDXI, SCIDX, IMD4), metabolic, liver, kidney and protein diseases and disorders (e.g., CROI encodes TTR, PALB, APOA1, APP, AAA, CVAP, ADI, GSN, FGA, LYZ, TTR, PALB, KRT18, KRT8, CIRH1A, NAIC, TEX292, KIAA1988, CFTR, ABCC7, CF, MRP7, SLC2A2, GLUT2, G6PC, G6PT, G6PT1, GAA, LAMP2, LAMPB, AGL, GDE, GBE1, GYS2, PYGL, PFKM, TCF1, HNF1A, MODY3, SCOD1, SCOl, CTNNB1, PDGFRL, PDGRL, PRLTS, AX1NI, AXIN, CTNNB1, TP53, P53, LFS1, IGF2R, MPRI, MET, CASP8, MCH5, UMOD, HNFJ, FJHN, MCKD2, ADMCKD2, PAH, PKU1, QDPR, DHPR, PTS, FCYT, PKHD1, ARPKD, PKD1, PKD2, PKD4, PKDTS, PRKCSH, G19P1, PCLD, SEC63), muscular / skeletal diseases and disorders (e.g., CROI encodes DMD, BMD, MYF6, LMNA, LMN1, EMD2, FPLD, CMDIA, HGPS, LGMDIB, LMNA, LMNI, EMD2, FPLD, CMDIA, FSHMD1A, FSHD1A, FKRP, MDC1C, LGMD2I, LAMA2, LAMM, LARGE, KIAA0609, MDC1D, FCMD, TTID, MYOT, CAPN3, CANP3, DYSF, LGMD2B, SGCG, LGMD2C, DMDA1, SCG3, SGCA, ADL, DAG2, LGMD2D, DMDA2, SGCB, LGMD2E, SGCD, SGD, LGMD2F, CMD1L, TCAP, LGMD2G, CMD1N, TRIM32, HT2A, LGMD2H, FKRP, MDCIC, LGMD21, TTN, CMD1G, TMD, LGMD2J, POMT1, CAV3, LGMD1C, SEPN1, SELN, RSMD1, PLEC1, PLTN, EBS1, LRP5, BMNDl, LRP7, LR3, OPPG, VBCH2, CLCN7, CLC7, OPTA2, OSTMI, GL, TCIRG1, TIRC7, OC116, OPTB1, VAPB, VAPC, ALS8, SMN1, SMA1, SMA2, SMA3, SMA4, BSCL2, SPG17, GARS, SMAD1, CMT2D, HEXB, IGHMBP2, SMUBP2, CATF1, SMARD1), neurological and neuronal diseases and disorders (e.g., CROI encodes SOD1, ALS2, STEX, FUS, TARDBP, VEGF (VEGF-a, VEGF-b, VEGF-c), APP, AAA, CVAP, ADI, APOE, AD2, PSEN2, AD4, STM2, APBB2, FE65LI, NOS3, PLAU, URK, ACE, DCPI, ACEI, MPO, PAC1PI, PAXIPIL, PTIP, A2M, BLMH, BMH, PSEN1, AD3, Mecp2, BZRAP1, MDGA2, Sema5A, Neurexin 1, GLOl, MECP2, RTT, PPMX, MRX16, MRX79, NLGN3, NLGN4, KIAA1260, AUTSX2, FMR2, FXR1, FXR2, mGLUR5, HD, IT15, PRNP, PRIP, JPH3, JP3, HDL2, TBP, SCA17, NR4A2, NURR1, NOT, TINUR, SNCAIP, TBP, SCA17, SNCA, NACP, PARK1, PARK4, DJI, PARK7, LRRK2, PARK8, PINK1, PARK6, UCHL1, PARK5, SNCA, NACP, PARKl, PARK4, PRKN, PARK2, PDJ, DBH, NDUFV2, MECP2, RTT, PPMX, MRX16, MRX79, CDKL5, STK9, MECP2, RTT, PPMX, MRX16,MRX79, x- Synuclein, DJ-1, Neuregulin-l (Nrgl), Erb4 (receptor for Neuregulin), Complexin-l (Cplxl), Tphl Tryptophan hydroxylase, Tph2, Tryptophan hydroxylase 2, Neurexin 1, GSK3, GSK3a, GSK3b, 5- HTT (Slc6a4), CONT, DRD (Drdla), SLC6A , DAOA, DTNBP1, Dao (Daol), APH-l(alpha and beta), Presenilin (Psenl), Nicastrin, (Ncstn), PEN-2, Nosl, Parpl, Natl, Nat2, HTT, SBMA / SMAX1 / AR, FXN / X25, ATX3, TXN, ATXN2, DMPK, Atrophin-1, Atnl, CBP, VLDLR, Atxn7, and AtxnlO), and ocular diseases and disorders (e.g., Aber, Ccl2, Cc2, cp (ceruloplasmin), Timp3, cathepsin-D, Vldlr, Ccr2, CRYAA, CRYA1, CRYBB2, CRYB2, PITX3, BFSP2, CP49, CP47, CRYAA, CRYAI, PAX6,AN2, MGDA, CRYBA1, CRYB1, CRYGC, CRYG3, CCL, LIM2, MP19, CRYGD, CRYG4, BFSP2, CP49, CP47, HSF4, CTM, HSF4, CTM, MIP, AQPO, CRYAB, CRYA2, CTPP2, CRYBB1, CRYGD, CRYG4, CRYBB2, CRYB2, CRYGC, CRYG3, CCL, CRYAA, CRYAI, GJA8, CX50, CAE1, GJA3, CX46, CZP3, CAE3, CCM1, CAM, KRIT1, APOA1, TGFBI, CSD2, CDGG1, CSD, BIGH3, CDG2, TACSTD2, TROP2, M1SI, VSX1, RINX, PPCD, PPD, KTCN, COL8A2, FECD, PPCD2, PIP5K3, CFD, KERA, CNA2, MYOC, TIGR, GLCIA, JO AG, GPOA, OPTN, GLC1E, FIP2, HYPL, NRP, CYP1BI, GLC3A, OPA1, NTG, NPG, CYP1BI, GLC3A, CRB1, RP12, CRX, CORD2, CRD, RPGRIPI, LCA6, CORD9, RPE65, RP20, AIPL1, LCA4, GUCY2D, GUC2D, LCA1, CORD6, RDH12, LCA3, ELOVL4, ADMD, STGD2, STGD3, RDS, RP7, PRPH2, PRPH, AVMD, AOFMD, and VMD2).

[0374] In some embodiments, the product coding region of the RNA payloads described herein encodes a factor that can affect the differentiation of a cell. As a non-limiting example, the expression of one or more of Oct4, Klf4, Sox2, c-Myc, L-Myc, dominant-negative p53, Nanog, Glisl, Lin28, TFIID, mir-302 / 367, or other miRNAs can cause the cell to become an induced pluripotent stem (iPS) cell.

[0375] In some embodiments, the product coding region of the RNA payloads described herein encodes a factor for transdifferentiating cells. Non-limiting examples of factors include: one or more of GATA4, Tbx5, Mef2C, Myocd, Hand2, SRF, Mespl, SMARCD3 for cardiomyocytes; Ascii, Nurrl, LmxlA, Bm2, Mytll, NeuroDl, FoxA2 for neural cells; and Hnf4a, Foxal, Foxa2 or Foxa3 for hepatic cells.

[0376] Additional product coding regions of the RNA payloads described herein are described elsewhere. A. Nucleic acid payloads

[0377] In various embodiments, the LNP compositions described herein can be used to deliver a nucleic acid or polynucleotide payload, e.g., a DNA HDR donor, a linear or circular mRNA, or a chimeric DNA / RNA guide.

[0378] In some embodiments, a LNP is capable of delivering a polynucleotide to a target cell, tissue, or organ. A polynucleotide, in its broadest sense of the term, comprises any compound and / or substance that is or can be incorporated into an oligonucleotide chain. Exemplary polynucleotides for use in accordance with the present disclosure include, but are not limited to, one or more of deoxyribonucleic acid (DNA), ribonucleic acid (RNA) including messenger mRNA (mRNA), hybrids thereof, RNAi-inducing agents, RNAi agents, siRNAs, shRNAs, miRNAs, antisense RNAs, ribozymes, catalytic DNA, RNAs that induce triple helix formation, aptamers, vectors, etc. RNAs useful in the compositions and methods described herein can be selected from the group consisting o...

Claims

CLAIMS 1. A compound of Formula (AX-AY)(AX-AY), or a pharmaceutically acceptable salt thereof, wherein: R1is selected from the group consisting of -OH, -OAc, -NR2, -N(R)RH,each R is independently -H or C1-C6 aliphatic; each RHis C1-C6 aliphatic-OH; each X1and XAare each independently a bond or optionally substituted C1-C6 aliphatic; each Y1is independently selected from the group consisting of, , , , , , ,, and a bond; wherein the bond marked with an "*" is attached to X1; each X2and X3is independently a bond or optionally substituted C1-C12 aliphatic; each Y2and Y3is independently selected from the group consisting of; wherein the bond marked with an "*" is attached to X2or X3; each X4and X5is independently a bond or optionally substituted C1-C6 aliphatic;each Y4and Y5is independently selected from the group consisting of a bond,; wherein the bond marked with an "*" is attached to X4or X5; each X6and X7is independently a bond or optionally substituted C1-C6 aliphatic; R2is -CH(OR6)(OR7), -CH(SR6)(SR7), -CH(R6)(R7), -CF(R6)(R7), -R10, optionally substituted C5-C18 aliphatic, or optionally substituted C1-C14 aliphatic-R10, wherein one or more methylene linkages of R2are each optionally and independently replaced with an optionally substituted C3-C8 cycloalkylenyl, phenyl, -O-, -NH-, -S-, -SS-, -C(O)-, - OC(O)O-, -OC(O)-, -NHC(O)-, or -C(O)O-; each R3is independently -CH(OR8)(OR9), -CH(SR8)(SR9), -CH(R8)(R9), -CF(R8)(R9), -R11, optionally substituted C5-C18 aliphatic, or optionally substituted C1-C14 aliphatic-R11, wherein one or more methylene linkages of R3are each optionally and independently replaced with an optionally substituted C3-C8 cycloalkylenyl, phenyl, -O-, -NH-, -S-, -SS- , -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or -C(O)O-; R6and R7are each independently optionally substituted -C1-C14 aliphatic, -R10, or optionally substituted -C1-C14 aliphatic-R10; wherein one or more methylene linkages of R6and R7are each optionally and independently replaced with an optionally substituted C3-C8 cycloalkylenyl, phenyl, -O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or - C(O)O-; R8and R9are each independently optionally substituted -C1-C14 aliphatic, -R11, or optionally substituted -C1-C14 aliphatic-R11; wherein one or more methylene linkages of R8and R9are each optionally and independently replaced with an optionally substituted C3-C8 cycloalkylenyl, phenyl, -O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or - C(O)O-; and each R10and R11is independently an optionally substituted cylic, bicyclic, bridged bicyclic, multicyclic or bridged multicyclic C4-C14 cycloalkyl or optionally substituted cylic, bicyclic, bridged bicyclic, multicyclic or bridged multicyclic 4-14 membered heterocyclyl, or two R10or two R11taken together form an optionally substituted bridged bicyclic or multicyclic C4-C14 cycloalkyl or optionally substituted bridged bicyclic or multicyclic 4-14 membered heterocyclyl; wherein one or more of X1, XA, X2, X3, X4, X5, X6, X7, R2and R3is optionally andindependently substituted with one or more substituents selected from -F, -Cl, -Br and -I.2.The compound of claim 1, wherein R1is selected from the group consisting of -OH, -NR2,3. The compound of claim 2, wherein R1is -NR2 or -N(R)RH; R of R1is C1-C6 aliphatic; and RHof R1is C1-C6 aliphatic-OH.

4. The compound of claim 1, wherein XA-R1is selected from the group consisting of5. The compound of claim 4, wherein Y1is selected from the group consisting of,and a bond; wherein the bond marked with an "*" is attached to X1.

6. The compound of claim 4, wherein X1is independently a bond or optionally substituted C1- C3aliphatic.

7. The compound of claim 4, wherein -X1-Y1-XA-R1is selected from the group consisting of8. The compound of claim 1, wherein each of R2and R3are independently selected from the group consisting of9. The compound of claim 1, wherein the compound is of Formula (AX-H)(AX-H), or a pharmaceutically acceptable salt thereof, wherein: R1is selected from the group consisting of -OH, -OAc, -NR2, -N(R)RH,,each R is independently -H or C1-C6 aliphatic; each RHis C1-C6 aliphatic-OH; each X1and XAare each independently a bond or optionally substituted C1-C6 aliphatic; each X2and X3is independently a bond or optionally substituted C1-C12 aliphatic; each X4and X5is independently a bond or optionally substituted C1-C6 aliphatic; each X6and X7is independently a bond or optionally substituted C1-C6 aliphatic; R6and R7are each independently optionally substituted -C1-C14 aliphatic, -R10, or optionally substituted -C1-C14 aliphatic-R10; wherein one or more methylene linkages of R6and R7are each optionally and independently replaced with an optionally substituted C3-C8 cycloalkylenyl, phenyl, -O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or - C(O)O-; R8and R9are each independently optionally substituted -C1-C14 aliphatic, -R11, or optionally substituted -C1-C14 aliphatic-R11; wherein one or more methylene linkages of R8and R9are each optionally and independently replaced with an optionally substituted C3-C8 cycloalkylenyl, phenyl, -O-, -NH-, -S-, -SS-, -C(O)-, -OC(O)O-, -OC(O)-, -NHC(O)-, or - C(O)O-; and each R10and R11is independently an optionally substituted cylic, bicyclic, bridged bicyclic, multicyclic or bridged multicyclic C4-C14 cycloalkyl or optionally substituted cylic, bicyclic, bridged bicyclic, multicyclic or bridged multicyclic 4-14 membered heterocyclyl, or two R10or two R11taken together form an optionally substituted bridged bicyclic or multicyclic C4-C14 cycloalkyl or optionally substituted bridged bicyclic or multicyclic 4-14 membered heterocyclyl; wherein one or more of X1, XA, X2, X3, X4, and X5, is optionally and independently substituted with one or more substituents selected from -F, -Cl, -Br and -I.10.The compound of claim 9, wherein X1is a bond.

11. The compound of claim 9, wherein X1is optionally substituted C1-C6 alkylene.12.The compound of claim 9, wherein X1is unsubstituted C1-C6 alkylene.

13. The compound of claim 12, wherein X1is unsubstituted C1-C2 alkylene.

14. The compound of any one of claims 9-13, wherein XAis a bond.

15. The compound of any one of claims 9-13, wherein XAis optionally substituted C1-C6 alkylene.

16. The compound of any one of claims 9-13, wherein XAis unsubstituted C1-C6 alkylene.

17. The compound of any one of claims 9-13, wherein XAis unsubstituted C1-C2 alkylene.

18. The compound of any one of claims 9-13, wherein R1is OH, -NR2, or.

19. The compound of any one of claims 9-13, wherein R1is -NR2 or.

20. The compound of claim 18, wherein R of R1is unsubstituted C1-C6 alkyl.

21. The compound of claim 18, wherein R of R1is unsubstituted C1-C4 alkyl.

22. The compound of claim 18, wherein R of R1is unsubstituted C1-C2 alkyl. 2 The compound of claim 9, wherein -X1-Y1-XA-R1is,24. The compound of claim 9, wherein X2is a bond.

25. The compound of claim 9, wherein X2is an optionally substituted C1-C4 alkylene.

26. The compound of claim 9, wherein X3is a bond.

27. The compound of claim 9, wherein X3is an optionally substituted C1-C4 alkylene.

28. The compound of claim 9, wherein X2and X3are the same.

29. The compound of claim 9, wherein X2and X3are both bond, methylene, ethylene, or propylene.

30. The compound of claim 9, wherein X2and X3are different.

31. The compound of claim 9, wherein X4is a bond.

32. The compound of claim 9, wherein X4is an optionally substituted C1-C4 alkylene.

33. The compound of claim 9, wherein X5is a bond.

34. The compound of claim 9, wherein X5is an optionally substituted C1-C4 alkylene.

35. The compound of claim 9, wherein X4and X5are the same.

36. The compound of claim 9, wherein X4and X5are both bond, methylene, ethylene, or propylene.

37. The compound of claim 9, wherein X4and X5are different.

38. The compound of claim 9, wherein R6and R7are each independently C1-C14 aliphatic, -R10, or -C1-C14 aliphatic-R10.

39. The compound of claim 38, wherein R6and R7are each independently C1-C14 alkyl.40.The compound of claim 38, wherein R6and R7are each independently C4-C10 alkyl.

41. The compound of claim 9, wherein R8and R9are each independently C1-C14 aliphatic, -R11, or -C1-C14 aliphatic-R11.42.The compound of claim 41, wherein R8and R9are each independently C1-C14 alkyl.

43. The compound of claim 41, wherein R8and R9are each independently C4-C10 alkyl.

44. The compound of claim 9, wherein R10is optionally substituted cyclic, bicyclic, bridged bicyclic, multicyclic or bridged multicyclic C4-C14 cycloalkyl or optionally substituted cylic, bicyclic, bridged bicyclic, multicyclic or bridged multicyclic 4-14 membered heterocyclyl.

45. The compound of claim 45, wherein R10is C4-C14 cycloalkyl.

46. The compound of claim 45, wherein R10is bridged bicyclic C6-C14 cycloalkyl.

47. The compound of claim 45, wherein R10is substituted bicyclic 6-14 membered heterocyclyl.4The compound of claim 45, wherein R10is.

49. The compound of claim 9, wherein R11is optionally substituted cyclic, bicyclic, bridged bicyclic, multicyclic or bridged multicyclic C4-C14 cycloalkyl or optionally substituted cylic, bicyclic, bridged bicyclic, multicyclic or bridged multicyclic 4-14 membered heterocyclyl.

50. The compound of claim 49, wherein R11is C4-C14 cycloalkyl.

51. The compound of claim 49, wherein R11is bridged C6-C14 cycloalkyl.

52. The compound of claim 49, wherein R11is substituted bicyclic 6-14 membered heterocyclyl.

53. The compound of claim 49, wherein R10is.

54. The compound of claim 1, wherein the compound is of Formula (AX-H’’)(AX-H’’), or a pharmaceutically acceptable salt thereof.

55. A compound selected from the group consisting of56. A pharmaceutical composition comprising a lipid nanoparticle, wherein the lipid nanoparticle comprises a ionizable lipid selected from a compound of any of claims 1-55.

57. The pharmaceutical composition comprising of claim 56, wherein the lipid nanoparticle further comprises: i) at least one structural lipid; ii) at least one phospholipid, non-ionizable lipid or zwitterionic lipid; and iii) at least one PEGylated lipid.

58. The pharmaceutical composition of claim 57, wherein the lipid nanoparticle further comprises an active agent.

59. The pharmaceutical composition of claim 58, wherein the active agent is a nucleic acid.

60. The pharmaceutical composition of claim 59, wherein the nucleic acid is a ribonucleic acid.

61. The pharmaceutical composition of claim 59, wherein the nucleic acid is a nucleic acid encoding a therapeutic protein.

62. The pharmaceutical composition of claim 61, wherein the nucleic acid encoding a therapeuticprotein comprises an open reading frame encoding a cancer antigen.

63. The pharmaceutical composition of claim 61, wherein the nucleic acid encoding a therapeutic protein comprises an open reading frame encoding an immune checkpoint modulator.

64. The pharmaceutical composition of any one of claims 61-63, wherein the nucleic acid encoding a therapeutic protein is an mRNA or a circular RNA (oRNA).

65. The pharmaceutical composition of claim 64, wherein the nucleic acid encoding a therapeutic protein is an mRNA.

66. The pharmaceutical composition of claim 65, wherein the mRNA comprises at least one motif selected from the group consisting of a stem loop, a chain terminating nucleoside, a polyA sequence, a polyadenylation signal, and a 5' cap structure.

67. The pharmaceutical composition of any one of claims 65-66, wherein the mRNA is at least 30 nucleotides in length.

68. The pharmaceutical composition of claim 65-67, wherein the mRNA is at least 300 nucleotides in length.

69. The pharmaceutical composition of claim 64, wherein the nucleic acid encoding a therapeutic protein is an oRNA.

70. The pharmaceutical composition of any one of claims 61-69, wherein the nucleic acid is suitable for a genome editing technique.

71. The pharmaceutical composition of claim 70, wherein the genome editing technique is CRISPR-Cas9 gene editing system.

72. The pharmaceutical composition of claim 70, wherein the genome editing technique is a Type V editor.

73. The pharmaceutical composition of any of claims 61-69, wherein the therapeutic protein is a CAR or TCR complex protein.

74. The pharmaceutical composition of claim 73, wherein the CAR or TCR complex protein comprises an antigen binding domain specific for an antigen selected from the group: CD 19, CD123, CD22, CD30, CD171, CS-1, C-type lectin-like molecule- 1, CD33, epidermal growth factor receptor variant III (EGFRvIII), disialoganglioside GD2, disaloganglioside GD3, TNF receptor family member, B cell maturation antigen (BCMA), Tn antigen ((Tn Ag) or(GalNAca-Ser / Thr)), prostate- specific membrane antigen (PSMA), Receptor tyrosine kinase- like orphan receptor 1 (ROR1), Fms-Like Tyrosine Kinase 3 (FLT3), Tumor-associated glycoprotein 72 (TAG72), CD38, CD44v6, Carcinoembryonic antigen (CEA), Epithelial cell adhesion molecule (EPCAM), B7H3 (CD276), KIT (CD 117), Interleukin- 13 receptor subunit alpha-2, mesothelin, Interleukin 11 receptor alpha (IL-l lRa), prostate stem cell antigen (PSCA), Protease Serine 21, vascular endothelial growth factor receptor 2 (VEGFR2), Lewis(Y) antigen, CD24, Platelet-derived growth factor receptor beta (PDGFR-beta), Stage-specific embryonic antigen-4 (SSEA-4), CD20, Folate receptor alpha, HER2, HER3, Mucin 1, cell surface associated (MUC1), epidermal growth factor receptor (EGFR), neural cell adhesion molecule (NCAM), Prostase, prostatic acid phosphatase (PAP), elongation factor 2 mutated (ELF2M), Ephrin B2, fibroblast activation protein alpha (FAP), insulin-like growth factor 1 receptor (IGF- I receptor), carbonic anhydrase IX (CAIX), Proteasome (Prosome, Macropain) Subunit, Beta Type, 9 (LMP2), glycoprotein 100 (gplOO), oncogene fusion protein consisting of breakpoint cluster region (BCR) and Abelson murine leukemia viral oncogene homolog 1 (Abl) (bcr-abl), tyrosinase, ephrin type- A receptor 2 (EphA2), Fucosyl GM1, sialyl Lewis adhesion molecule (sLe), ganglioside GM3, transglutaminase 5 (TGS5), high molecular weight-melanoma- associated antigen (HMWMAA), o-acetyl-GD2 ganglioside (0AcGD2), Folate receptor beta, tumor endothelial marker 1 (TEM1 / CD248), tumor endothelial marker 7 -related (TEM7R), claudin 6 (CLDN6), claudin 18.2 (CLDN18.2), thyroid stimulating hormone receptor (TSHR), G protein-coupled receptor class C group 5, member D (GPRC5D), chromosome X open reading frame 61 (CXORF61), CD97, and CD179a.

75. The pharmaceutical composition of claim 73, wherein the CAR or TCR complex protein comprises BCMA, CD19, or CD20.

76. The pharmaceutical composition of claim 60, wherein the ribonucleic acid is at least one ribonucleic acid selected from the group consisting of a small interfering RNA (siRNA), an asymmetrical interfering RNA (aiRNA), a microRNA (miRNA), a Dicer-substrate RNA (dsRNA), a small hairpin RNA (shRNA), a messenger RNA (mRNA), and a long non-coding RNA (lncRNA).

77. The pharmaceutical composition of any one of claims 57-76, wherein the lipid component of the nanoparticle composition comprises about 30 mol % to about 40 mol % ionizable lipid; about 35 mol % to about 45 mol % phospholipid; about 20 mol % to about 30 mol % structural lipid; and about 0.5 mol % to about 5 mol % of PEG lipid,provided that the total mol % does not exceed 100%.

78. The pharmaceutical composition of any one of claims 57-76, wherein the lipid component of the nanoparticle composition comprises about 25 mol % to about 45 mol % ionizable lipid; about 35 mol % to about 50 mol % phospholipid; about 10 mol % to about 25 mol % structural lipid; and about 1 mol% to about 5 mol% of PEG lipid, provided that the total mol % does not exceed 100%.

79. The pharmaceutical composition of any one of claims 57-76, wherein the lipid component of the nanoparticle composition comprises about 30 mol % to about 40 mol % ionizable lipid; about 35 mol % to about 45 mol % phospholipid; about 20 mol % to about 30 mol % structural lipid; and about 0.5 mol % to about 5 mol % of PEG lipid provided that the total mol % does not exceed 100%.

80. The pharmaceutical composition of any one of claims 57-76, wherein the lipid component of the nanoparticle composition comprises about 30 mol % to about 36 mol % ionizable lipid; about 35 mol % to about 45 mol % phospholipid; about 22 mol % to about 27 mol % structural lipid; and about 0.5 mol% to about 3.5 mol% of PEG lipid, provided that the total mol % does not exceed 100%.

81. The pharmaceutical composition of any one of claims 57-76, wherein the lipid component of the nanoparticle composition comprises about 33 mol % ionizable lipid; about 40 mol % phospholipid; about 25 mol % structural lipid; and about 2 mol% of PEG lipid, provided that the total mol % does not exceed 100%.

82. The pharmaceutical composition of any one of claims 57-81, wherein the at least one structural lipid is selected from cholesterol, fecosterol, fucosterol, beta sitosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, cholic acid, sitostanol, litocholic acid,tomatine, ursolic acid, alpha-tocopherol, Vitamin D3, Vitamin D2, Calcipotriol, botulin, lupeol, oleanolic acid, beta-sitosterol-acetate and any combinations thereof.

83. The pharmaceutical composition of any one of claims 57-82, wherein the at least one phospholipid is selected from 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2- dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-dilinoleoyl-sn-glycero-3- phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero-phosphocholine (DMPC), 1.2-dioleoyl- sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-diundecanoyl-sn-glycero-phosphocholine (DUPC), 1-palmitoyl-2-oleoyl-sn-glycero-3- phosphocho line (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 Diether PC), 1-oleoyl-2-cholesterylhemisuc cinoyl-sn-glycero-3-phosphocholine (OChemsPC), 1- hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dilinolenoyl-sn-glycero-3- phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl-sn- glycero-3-phosphocholine, 1,2-diphytanoylsn-glycero-3-phosphoethanolamine (ME 16.0 PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3- phosphoethanolamine, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2- diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1,2-didocosahexaenoyl-sn-glycero-3- phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG), sodium (S)-2-ammonio-3-((((R)-2-(oleoyloxy)-3- (stearoyloxy)propoxy)oxidophosphoryl)oxy)propanoate (L-α-phosphatidylserine; Brain PS), dimyristoyl phosphatidylcholine (DMPC), dimyristoyl phosphoethanolamine (DMPE), dimyristoylphosphatidylglycerol (DMPG), dioleoyl-phosphatidylethanolamine4-(N- maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dioleoylphosphatidylglycerol (DOPG), 1,2-dioleoyl-sn-glycero-3-(phospho-L-serine) (DOPS), acell-fusogenicphospholipid (DPhPE), dipalmitoylphosphatidylethanolamine (DPPE), 1,2-Dielaidoyl-sn- phosphatidylethanolamine (DEPE), dipalmitoylphosphatidylglycerol (DPPG), dipalmitoylphosphatidylserine (DPPS), distearoylphosphatidylcholine (DSPC), distearoyl- phosphatidyl-ethanolamine (DSPE), distearoyl phosphoethanolamineimidazole (DSPEI), 1,2- diundecanoyl-sn-glycero-phosphocholine (DUPC), egg phosphatidylcholine (EPC), 1,2- dioleoyl-sn-glycero-3-phosphate (18:1 PA; DOPA), ammonium bis((S)-2-hydroxy-3- (oleoyloxy)propyl) phosphate (18:1 DMP; LBPA), 1,2-dioleoyl-sn-glycero-3-phospho-(1’- myo-inositol) (DOPI; 18:1 PI), 1,2-distearoyl-sn-glycero-3-phospho-L-serine (18:0 PS), 1,2- dilinoleoyl-sn-glycero-3-phospho-L-serine (18:2 PS), 1-palmitoyl-2-oleoyl-sn-glycero-3- phospho-L-serine (16:0-18:1 PS; POPS), 1-stearoyl-2-oleoyl-sn-glycero-3-phospho-L-serine (18:0-18:1 PS), 1-stearoyl-2-linoleoyl-sn-glycero-3-phospho-L-serine (18:0-18:2 PS), 1- oleoyl-2-hydroxy-sn-glycero-3-phospho-L-serine (18:1 Lyso PS), 1-stearoyl-2-hydroxy-sn- glycero-3-phospho-L-serine (18:0 Lyso PS), and sphingomyelin.

84. The pharmaceutical composition of any one of claims 57-83, wherein the at least one PEGylated lipid is selected from (R)-2,3-bis(octadecyloxy)propyl-1- (methoxypoly(ethyleneglycol)2000)propylcarbamate, PEG-S-DSG, PEG-S-DMG, PEG-PE, PEG-PAA, PEG-OH DSPE C18, PEG-DSPE, PEG-DSG, PEG-DPG, PEG-DOMG, PEG- DMPE Na, PEG-DMPE, PEG-DMG2000, PEG-DMG C14, PEG-DMG 2000, PEG-DMG, PEG-DMA, PEG-Ceramide C16, PEG-C-DOMG, PEG-c-DMOG, PEG-c-DMA, PEG-cDMA, PEGA, PEG750-C-DMA, PEG400, PEG2k-DMG, PEG2k-C11, PEG2000-PE, PEG2000P, PEG2000-DSPE, PEG2000-DOMG, PEG2000-DMG, PEG2000-C-DMA, PEG2000, PEG200, PEG(2k)-DMG, PEG DSPE C18, PEG DMPE C14, PEG DLPE C12, PEG Click DMG C14, PEG Click C12, PEG Click C10, N(Carbonyl-methoxypolyethylenglycol-2000)- l,2-distearoyl-sn-glycero3-phosphoethanolamine, Myrj52, mPEG-PLA, MPEG-DSPE, mPEG3000-DMPE, MPEG-2000-DSPE, MPEG2000-DSPE, mPEG2000-DPPE, mPEG2000- DMPE, mPEG2000-DMG, mDPPE-PEG2000, l,2-distearoyl-sn-glycero-3- phosphoethanolamine-PEG2000, HPEG-2K-LIPD, Folate PEG-DSPE, DSPE-PEGMA 500, DSPE-PEGMA, DSPE-PEG6000, DSPE-PEG5000, DSPE-PEG2K-NAG, DSPE-PEG2k, DSPE-PEG2000maleimide, DSPE-PEG2000, DSPE-PEG, DSG-PEGMA, DSG-PEG5000, DPPE-PEG-2K, DPPE-PEG, DPPE-mPEG2000, DPPE-mPEG, DPG-PEGMA, DOPE- PEG2000, DMPE-PEGMA, DMPE-PEG2000, DMPE-Peg, DMPE-mPEG2000, DMG- PEGMA, DMG-PEG2000, DMG-PEG, distearoyl-glycerol-polyethyleneglycol, Cl8PEG750, CI8PEG5000, CI8PEG3000, CI8PEG2000, CI6PEG2000, CI4PEG2000, C18-PEG5000, C18PEG, C16PEG, C16 mPEG (polyethylene glycol) 2000 Ceramide, C14-PEG-DSPE200, C14-PEG2000, C14PEG2000, C14-PEG 2000, C14-PEG, C14PEG, 14:0-PEG2KPE, 1,2- distearoyl-sn-glycero-3-phosphoethanolamine-PEG2000, (R)-2,3-bis(octadecyloxy)propyl-1- (methoxypoly(ethyleneglycol)2000)propylcarbamate, (PEG)-C-DOMG, PEG-C-DMA, and DSPE-PEG-X.

85. The pharmaceutical composition of any one of claims 57-84 wherein: the PEG lipid is PEG2k-DMG; the structural lipid is cholesterol; and the phospholipid is a ESM or DSPC or a mixture thereof.

86. The pharmaceutical composition of any one of claims 57-76, wherein the lipid component of the nanoparticle composition comprises about 33 mol % ionizable lipid; about 40 mol % ESM; about 25 mol % cholesterol; and about 2 mol% of PEG2k-DMG lipid,provided that the total mol % does not exceed 100%.

87. The pharmaceutical composition of any one of claims 57-76, wherein the lipid component ofthe nanoparticle composition comprises about 33 mol % ionizable lipid; about 40 mol % DSPC; about 25 mol % cholesterol; and about 2 mol% of PEG2k-DMG lipid, provided that the total mol % does not exceed 100%.

88. The pharmaceutical composition of any one of claims 57-76, wherein the lipid component ofthe nanoparticle composition comprises about 33 mol % ionizable lipid; about 20 mol % DSPC; about 20 mol % ESM; about 25 mol % cholesterol; and about 2 mol% of PEG2k-DMG lipid, provided that the total mol % does not exceed 100%.