Lipid nanoparticle and use thereof

Abstract

Provided is a lipid nanoparticle, which comprises an active component, an ionizable lipid, an auxiliary lipid, a structural lipid, and a polymer lipid. The polymer lipid accounts for 0-0.8 mol% of the total lipid existing in the lipid nanoparticle. The reduction in the content of the polymer lipid in the lipid nanoparticle can change the delivery specificity of the lipid nanoparticle, such that the lipid nanoparticle can target the spleen or immune cells.

Description

Lipid nanoparticles and applications thereof TECHNICAL FIELD

[0001] The present application belongs to the field of biotechnology, and relates to a kind of lipid nanoparticles and its application. BACKGROUND

[0002] Nucleic acids (such as mRNA, siRNA, etc.) have become a new class of drugs for preventing and treating various diseases. In order for nucleic acids to function in vivo, a safe, effective and stable delivery system is needed to protect them from degradation and allow cell uptake and release. Common delivery systems for delivering nucleic acids include lipid nanoparticles (LNPs). Lipid nanoparticles generally contain ionizable lipids, structural lipids, helper lipids and PEG lipids.

[0003] Traditional lipid nanoparticles have significant enrichment in the liver after intravenous injection, but have low delivery efficiency to other organs (such as lungs, kidneys, spleen, etc.). The spleen is the largest immune organ in the body, accounting for 25% of the total amount of lymphatic tissue in the body, and is the center of cellular and humoral immunity, playing a role in anti-tumor and other mechanisms through various mechanisms. Therefore, it has important application value to achieve spleen-targeted delivery of nucleic acid drugs.

[0004] Currently, the targeting delivery to the spleen is mainly achieved by optimizing the formulation of lipid nanoparticles and screening new lipids. For example, Cheng, Qiang et al. ("Selective organ targeting (SORT) nanoparticles for tissue-specific mRNA delivery and CRISPR-Cas gene editing." Nature nanotechnology 15.4 (2020): 313-320.) added a fifth component (named as SORT molecule) to the traditional four-component liver-targeting lipid nanoparticles to prepare a five-component lipid nanoparticle system. When the SORT molecule is a negatively charged lipid (such as 18PA), the lipid nanoparticles specifically deliver mRNA into the spleen for expression. For example, Fenton, Owen S. et al. ("Synthesis and biological evaluation of ionizable lipid materials for the in vivo delivery of messenger RNA to B lymphocytes." Advanced Materials 29.33 (2017): 1606944.) screened and obtained a spleen-targeting ionizable lipid (OF-Deg-Lin). Based on OF-Deg-Lin, LNPs can target mRNA delivery to the spleen. SUMMARY

[0005] The present disclosure provides a use of a lipid nanoparticle (LNPs) in the preparation of a medicament for targeting the spleen or immune cells, the LNPs comprising an active ingredient, an ionizable lipid, a helper lipid, a structural lipid, and a polymer lipid, the polymer lipid accounting for 0 mol% to 0.8 mol% of the total lipid present in the LNPs.

[0006] The present disclosure also provides a use of a lipid nanoparticle (LNPs) in the preparation of a medicament for targeting the spleen or immune cells, the LNPs comprising an active ingredient, an ionizable lipid, a helper lipid, a structural lipid, and a polymer lipid, the particle size of the LNPs being 130 nm to 500 nm.

[0007] In some embodiments, the particle size of the LNPs is 130 nm to 500 nm.

[0008] In some embodiments, the polymer lipid accounts for 0 mol% to 0.8 mol% of the total lipid present in the LNPs.

[0009] In some embodiments, the polymer lipid is a PEG lipid.

[0010] In some embodiments, the PEG lipid is selected from one or more of DMG-PEG, DSG-PEG, and DPG-PEG.

[0011] In some embodiments, the LNPs have a particle size of 150 nm to 450 nm, 150 nm to 400 nm, 150 nm to 390 nm, 150 nm to 380 nm, 150 nm to 370 nm, 150 nm to 360 nm, 150 nm to 350 nm, 150 nm to 340 nm, 150 nm to 330 nm, 150 nm to 320 nm, 150 nm to 310 nm, 150 nm to 300 nm, 150 nm to 290 nm, 150 nm to 280 nm, 150 nm to 270 nm, 150 nm to 260 nm, 150 nm to 250 nm, 150 nm to 240 nm, 150 nm to 230 nm, 150 nm to 210 nm, 150 nm to 200 nm, 160 nm to 320 nm, 160 nm to 310 nm, 160 nm to 300 nm, 160 nm to 290 nm, 160 nm to 280 nm, 160 nm to 270 nm, 160 nm to 260 nm, 160 nm to 250 nm, 160 nm to 240 nm, 160 nm to 230 nm, 160 nm to 230 nm, 160 nm to 210 nm, 160 nm to 200 nm, 180 nm to 320 nm, 180 nm to 310 nm, 180 nm to 300 nm, 180 nm to 290 nm, 180 nm to 280 nm, 180 nm to 270 nm, 180 nm to 260 nm, 180 nm to 250 nm, 180 nm to 240 nm, 180 nm to 230 nm, 180 nm to 230 nm, 180 nm to 210 nm, 180 nm to 200 nm, 190 nm to 320 nm, 190 nm to 310 nm, 190 nm to 300 nm, 190 nm to 290 nm, 190 nm to 280 nm, 190 nm to 270 nm, 190 nm to 260 nm, 190 nm to 250 nm, 190 nm to 240 nm, 190 nm to 230 nm, 190 nm to 230 nm, 190 nm to 210 nm, 190 nm to 200 nm, 200 nm to 320 nm, 200 nm to 310 nm, 200 nm to 300 nm, 200 nm to 290 nm, 200 nm to 280 nm, 200 nm to 270 nm, 200 nm to 260 nm, 200 nm to 250 nm, 200 nm to 240 nm, 200 nm to 230 nm, or 200 nm to 210 nm.

[0012] In some embodiments, the polymeric lipid comprises 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.7 mol%, 0.01 mol% to 0.6 mol%, 0.01 mol% to 0.5 mol%, 0.01 mol% to 0.4 mol%, 0.01 mol% to 0.3 mol%, 0.01 mol% to 0.2 mol%, 0.01 mol% to 0.1 mol%, 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.7 mol%, 0.05 mol% to 0.6 mol%, 0.05 mol% to 0.5 mol%, 0.05 mol% to 0.4 mol%, 0.05 mol% to 0.3 mol%, 0.05 mol% to 0.25 mol%, 0.05 mol% to 0.2 mol%, 0.05 mol% to 0.15 mol%, 0.05 mol% to 0.1 mol%, 0.1 mol% to 0.8 mol%, 0.1 mol% to 0.7 mol%, 0.1 mol% to 0.6 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.4 mol%, 0.1 mol% to 0.3 mol%, 0.1 mol% to 0.2 mol%, 0.1 mol% to 0.15 mol%, 0.15 mol% to 0.8 mol%, 0.15 mol% to 0.7 mol%, 0.15 mol% to 0.6 mol%, 0.15 mol% to 0.5 mol%, 0.15 mol% to 0.4 mol%, 0.15 mol% to 0.3 mol%, 0.15 mol% to 0.25 mol%, 0.2 mol% to 0.8 mol%, 0.2 mol% to 0.7 mol%, 0.2 mol% to 0.6 mol%, 0.2 mol% to 0.5 mol%, 0.2 mol% to 0.4 mol%, or 0.2 mol% to 0.35 mol% of the total lipid present in the LNPs.

[0013] In some embodiments, the ionizable lipid comprises 30 mol% to 60 mol% of the total lipid present in the LNPs, the helper lipid comprises 0 mol% to 30 mol% of the total lipid present in the LNPs, and the structural lipid comprises 10 mol% to 60 mol% of the total lipid present in the LNPs.

[0014] In some embodiments, the helper lipid is selected from one or more of DSPC, DOPE, DOPC, and DOPS.

[0015] In some embodiments, the structural lipid is cholesterol.

[0016] In some embodiments, the immune cells comprise one or more of B cells, T cells, NK cells, DC cells, neutrophils, eosinophils, basophils, mast cells, monocytes, and macrophages.

[0017] In some embodiments, the immune cells are B cells, T cells, or NK cells.

[0018] In some embodiments, the active ingredient is a nucleic acid, a protein, a polypeptide, or a small molecule.

[0019] In some embodiments, the medicament is for preventing or treating a disease associated with immune cells.

[0020] In some embodiments, the medicament is for preventing or treating an autoimmune disease.

[0021] In some embodiments, the active ingredient is a nucleic acid that suppresses an autoimmune response.

[0022] In some embodiments, the medicament is for preventing or treating a T cell tumor or a B cell tumor.

[0023] In some embodiments, the active ingredient is a nucleic acid that suppresses growth of a T cell tumor or a B cell tumor or kills T cell tumor cells or B cell tumor cells.

[0024] In some embodiments, the medicament is for preventing or treating a tumor or a cancer, and the active ingredient is a tumor-associated antigen or a nucleic acid encoding a tumor-associated antigen.

[0025] In some embodiments, the medicament is for preventing or treating an infectious disease, and the active ingredient is an antigen associated with the infectious disease or a nucleic acid encoding an antigen associated with the infectious disease.

[0026] In some embodiments, the nucleic acid is an mRNA.

[0027] The present disclosure also provides use of a lipid nanoparticle (LNP) in the manufacture of a genetically edited immune cell, the LNP being as defined in any of the preceding embodiments.

[0028] In some embodiments, the genetically edited immune cell is a B cell, a T cell, or a NK cell.

[0029] In some embodiments, the active ingredient is a nucleic acid required for the genetic editing.

[0030] In some embodiments, the active ingredient is a gRNA and a nucleic acid encoding a Cas protein.

[0031] In some embodiments, the active ingredient is a gRNA, a nucleic acid encoding a Cas protein and a donor DNA.

[0032] The present disclosure also provides the use of a lipid nanoparticle (LNPs) as defined in any of the preceding embodiments for the manufacture of a modified immune cell.

[0033] In some embodiments, the modified immune cell is a CAR-T cell, a TCR-T cell or a CAR-NK cell.

[0034] In some embodiments, the active ingredient is an mRNA encoding a CAR or a TCR.

[0035] The present disclosure also provides a method of increasing the amount of an active ingredient in the spleen or immune cells, the method comprising the step of contacting LNPs as defined in any of the preceding embodiments with the spleen or immune cells.

[0036] In some embodiments, the active ingredient is an mRNA.

[0037] In some embodiments, the method is for increasing the amount of an active ingredient delivered into the spleen or immune cells.

[0038] The present disclosure also provides a method of increasing the ratio of an active ingredient in the spleen and liver, the method comprising the step of contacting LNPs as defined in any of the preceding embodiments with the spleen or immune cells.

[0039] In some embodiments, the LNPs have a spleen or immune cell delivery preference, the method is for increasing the spleen / liver delivery ratio of an active ingredient.

[0040] In some embodiments, the active ingredient is an mRNA.

[0041] In some embodiments, the method is for increasing the expression of the mRNA in the spleen or immune cells, or the method is for increasing the spleen / liver delivery ratio of the mRNA.

[0042] In some embodiments, the method is performed in vivo.

[0043] In some embodiments, the method comprises the step of administering the LNPs to a subject.

[0044] The present disclosure also provides a lipid nanoparticle (LNPs) as defined in any of the preceding embodiments.

[0045] In some embodiments, the active ingredient is an mRNA.

[0046] The present disclosure also provides a composition for preparing the LNPs for use described above.

[0047] In some embodiments, the active ingredient is an mRNA.

[0048] DETAILED DESCRIPTION

[0049] I. DEFINITIONS

[0050] All patents, patent applications, scientific publications, manufacturer's specifications and handbooks, etc., cited herein are hereby incorporated by reference in their entirety. Nothing herein is to be construed as an admission that the patent disclosure is not entitled to antedate such disclosure.

[0051] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as those commonly understood by one of ordinary skill in the art to which this disclosure belongs. Also, all patents, patent applications, scientific publications, manufacturer's specifications and handbooks, etc., cited herein are hereby incorporated by reference in their entirety. Nothing herein is to be construed as an admission that the patent disclosure is not entitled to antedate such disclosure.

[0052] As used herein, the expressions "comprise", "comprising", "include", "including", "contain", "containing", and "have", "having", are open-ended, and do not exclude additional, unrecited elements, steps, or components. The expression "consisting of" excludes any element, step, or component not specified. The expression "consisting essentially of" means that the scope of the expression is limited to the specified elements, steps, or components, plus optional elements, steps, or components that do not materially affect the basic and novel characteristics of the claimed subject matter. It is understood that the expressions "consisting essentially of" and "consisting of" are encompassed within the meaning of the expression "comprising".

[0053] As used herein, the singular forms "a", "an" and "the" and the like carry the meaning of the singular as well as the plural, unless the context clearly dictates otherwise. The terms "one or more" or "at least one" encompass 1, 2, 3, 4, 5, 6, 7, 8, 9, or more. The terms "one or more" or "at least one" encompass 1, 2, 3, 4, 5, 6, 7, 8, 9, or more. The terms "at least two" encompass 2, 3, 4, 5, 6, 7, 8, 9, or more. The terms "at least two" encompass 2, 3, 4, 5, 6, 7, 8, 9, or more.

[0054] Numerical ranges as described herein are to be understood as encompassing any and all subranges that fall within the given range. For example, a range of "1 to 10" should be interpreted to include not only the explicitly enumerated values (that is, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10), but also values that are in between each of the explicit values. Thus, for a range of 1 to 10, this would include a range of 2 to 8, 3 to 7, 4 to 6, and 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10, etc. This principle can also apply to ranges with a lower limit of zero. Thus, a range of 0 to 10 should be interpreted to include not only the explicitly enumerated values (that is, 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10), but also values that are in between each of the explicit values.

[0055] As used herein, the terms "and / or", "any of", and "or" are interchangeable. These terms can express, in particular, any combination. For example, the following phrases "A, B, and / or C" or "A, B, C, or any of the same" mean any one of the following: "A alone; B alone; C alone; A and B; B and C; A and C; and A, B, and C".

[0056] In this document, "nucleotides" include deoxyribonucleotides, ribonucleotides, deoxyribonucleotide derivatives, and ribonucleotide derivatives. As used herein, "ribonucleotides" are the constituent materials of ribonucleic acid (RNA), consisting of one molecule of base, one molecule of five-carbon sugar, and one molecule of phosphate, which refers to a nucleotide having a hydroxyl group at the 2' position of the β-D-ribofuranosyl group, and "deoxyribonucleotides" are the constituent materials of deoxyribonucleic acid (DNA), also consisting of one molecule of base, one molecule of five-carbon sugar, and one molecule of phosphate, which refers to a nucleotide having a hydrogen instead of a hydroxyl group at the 2' position of the β-D-ribofuranosyl group, which is the main chemical component of chromosomes.

[0057] A nucleotide is generally referred to by a single letter representing the base therein: "A" or "A nucleotide" refers to an adenine deoxyribonucleotide or an adenine ribonucleotide containing adenine, "C" or "C nucleotide" refers to a cytosine deoxyribonucleotide or a cytosine ribonucleotide containing cytosine, "G" or "G nucleotide" refers to a guanine deoxyribonucleotide or a guanine ribonucleotide containing guanine, "U" or "U nucleotide" refers to a uracil ribonucleotide containing uracil, and "T" or "T nucleotide" refers to a thymine deoxyribonucleotide containing thymine.

[0058] As used herein, the term "nucleic acid" generally refers to any compound comprising a polymer of deoxyribonucleotides (deoxyribonucleic acid, DNA for short) or a polymer of ribonucleotides (ribonucleic acid, RNA for short), or a combination thereof. In addition, nucleic acid herein also includes derivatives of nucleic acid. The term "derivative of nucleic acid" includes chemical derivatization of nucleic acid on the base, sugar, or phosphate of the nucleotide, as well as nucleic acid containing non-natural nucleotides and nucleotide analogs. Furthermore, in this context, nucleic acid can be in the form of a single-stranded or double-stranded linear or covalently closed circular molecule.

[0059] A polynucleotide can comprise one segment or multiple segments (nucleic acid fragments) (e.g., 1, 2, 3, 4, 5, 6, 7, 8 segments). For example, a polynucleotide can comprise a segment encoding a protein or polypeptide of interest. In particular embodiments, a polynucleotide can comprise a segment encoding a protein or polypeptide of interest and a regulatory segment (including but not limited to segments for transcriptional regulation and translational regulation). In an embodiment, the regulatory segment comprises polynucleotide corresponding to one or more of the following elements that function in regulation: a promoter, a 5' untranslated region (5'-UTR), a 3' untranslated region (3'-UTR), and a poly(A) tail.

[0060] The term "promoter" refers to a polynucleotide located upstream from the 5' end of a coding region of a gene that contains conserved sequences required for specific binding of RNA polymerase and initiation of transcription, and which activates RNA polymerase to bind to the template DNA and have specificity for initiation of transcription. A promoter can be derived from one or more of viruses, bacteria, fungi, plants, insects, and animals. Representative examples of promoters include a bacteriophage T7 promoter, a bacteriophage T3 promoter, an SP6 promoter, a lac operator-promoter, a tac promoter, an SV40 late promoter, an SV40 early promoter, an RSV-LTR promoter, a CMV IE promoter, an SV40 early promoter, or an SV 40 late promoter and a CMV IE promoter.

[0061] As used herein, the term "5' untranslated region" or "5'-UTR" can refer to an RNA sequence in mRNA that is upstream of the coding sequence and is not translated into protein. A 5'-UTR in a gene typically begins at the transcription start site and ends with a nucleotide upstream of the translation start codon in the coding sequence. The 5'-UTR can contain elements that control gene expression, such as ribosome binding sites, 5'-terminal oligopyrimidine bundles, and translation initiation signals such as the Kozak sequence. mRNA can undergo post-transcriptional modification by adding a 5' cap. Therefore, the 5'-UTR in mature mRNA can also refer to the RNA sequence between the 5' cap and the start codon.

[0062] As used herein, the term "3' untranslated region" or "3'-UTR" can refer to an RNA sequence in mRNA that is downstream of the coding sequence and is not translated into a protein. The 3'-UTR in mRNA is located between a stop codon and a poly(A) sequence of the coding sequence, for example, starting from a nucleotide downstream of the stop codon and ending at a nucleotide upstream of the poly(A) sequence.

[0063] As used herein, the terms “poly(A) nucleotide,” “polyA,” “poly(A) sequence,” and “poly(A) tail” are used interchangeably. Naturally occurring poly(A) sequences typically consist of adenine ribonucleotides. Poly(A) sequences are usually located at the 3' end of mRNA, such as the 3' end (downstream) of a 3'-UTR.

[0064] As used herein, the term "5'-cap structure" refers to a structure that is typically located at the 5' end of mature mRNA. In some embodiments, the 5'-cap structure is linked to the 5' end of the mRNA via a 5'-5'-triphosphate bond. The 5'-cap structure is typically formed from modified (e.g., methylated) ribonucleotides, particularly guanine nucleotide derivatives. For example, m... 7 GpppN (cap0, or "cap0") is formed when the 5' phosphate group of hnRNA binds to m under the action of guanylate transferase. 7 The 5'-phosphate group of GTP forms a 5',5'-phosphodiester bond to form a cap structure, where N is the terminal 5' nucleotide of the nucleic acid carrying the 5'-cap structure. In some embodiments, the 5'-cap structure includes, but is not limited to, cap 0, cap 1 (a cap structure formed by further methylation of the 2'-OH of the first nucleotide glycosyl group of hnRNA on the basis of cap 0, or "cap1"), cap 2 (a cap structure formed by further methylation of the 2'-OH of the second nucleotide glycosyl group of hnRNA on the basis of cap 1, or "cap2"), cap 4, cap 0 analogue, cap 1 analogue, cap 2 analogue, or cap 4 analogue.

[0065] As used herein, the term "expression" includes transcription and / or translation of a nucleotide sequence. Thus, expression can involve the production of a transcript and / or a polypeptide. The term "transcription" relates to the process of transcribing the genetic code in a DNA sequence into RNA (transcript). The term "in vitro transcription" refers to the synthesis of RNA, in particular mRNA, in vitro in a system free of cells, for example in an appropriate cell extract (see, e.g., Pardi N., Muramatsu H., Weissman D., Kariko K. (2013). In: Rabinovich P. (eds) Synthetic Messenger RNA and Cell Metabolism Modulation. Methods in Molecular Biology (Methods and Protocols), vol 969. Humana Press, Totowa, NJ.). A vector that can be used for the production of a transcript is also referred to as "transcription vector", wherein regulatory sequences required for transcription are comprised. The term "transcription" encompasses "in vitro transcription".

[0066] As used herein, the term "polypeptide" refers to a polymer comprising two or more amino acids covalently linked by peptide bonds. A "protein" can comprise one or more polypeptides, wherein the polypeptides interact with each other by covalent or non-covalent means.

[0067] In the context of the present application, the term "plasmid" generally refers to a circular DNA molecule, but the term can also encompass linearized DNA molecules. Specifically, the term "plasmid" also encompasses molecules obtained by linearizing a circular plasmid, e.g., by digesting the circular plasmid with a restriction enzyme, thereby transforming the circular plasmid molecule into a linear molecule. Plasmids can replicate, i.e., amplify in a cell independent of the genetic information stored as chromosomal DNA, and can be used for cloning, i.e., for amplifying genetic information in bacterial cells. In some embodiments, the DNA plasmid is a medium- or high-copy plasmid. In some embodiments, the DNA plasmid is a high-copy plasmid. Examples of such high-copy plasmids include, e.g., pUC and pTZ plasmids or any other plasmid comprising an origin of replication supporting high-copy plasmids (e.g., pMB1, pCoIE1).

[0068] As used herein, the term "antigen" generally refers to a substance that can be recognized by the immune system, preferably by the adaptive immune system, and is capable of triggering an antigen-specific immune response (e.g. formation of antibodies and / or antigen-specific T cells). Typically, an antigen can be or can comprise a peptide or protein that can be presented by MHC to T cells. In the sense of the present disclosure, an antigen can be the translation product of a provided nucleic acid (e.g. RNA, RNA molecule, DNA herein). Furthermore, fragments, variants and derivatives of peptides or proteins derived from a peptide or protein comprising at least one epitope or antigen (e.g. tumor antigen, viral antigen, bacterial antigen, protozoal antigen) can be understood as an antigen.

[0069] As used herein, the term "treatment" and the like, are used herein to generally mean obtaining a desired pharmacologic and / or physiologic effect. Thus, treatment according to the present application can relate to the treatment of the state of a disease, but can also relate to a prophylactic treatment in the sense of a complete or partial prevention of a disease or symptoms thereof. In some embodiments, the term "treatment" is to be understood as being therapeutic in the sense of a partial or complete cure of a disease and / or adverse effects and / or symptoms attributable to the disease. Treatment can also be a prophylactic or preventative treatment, i.e. a measure taken to prevent a disease, e.g. to prevent the onset of an infection and / or disease.

[0070] As used herein, the terms "subject" and "patient" can be used interchangeably. In certain embodiments, a subject is a mammal, such as a human, a non-human primate (e.g., simians, chimpanzees, monkeys, and orangutans), a domestic animal (including dogs and cats and farm animals (e.g., horses, cows, pigs, sheep, and goats)), or other mammal. Other mammals include, but are not limited to, mice, rats, guinea pigs, hamsters, rabbits, and the like. In a particular embodiment, a subject is a human. In one embodiment, a subject is a mammal (e.g., a human) having a genetic disease, an orphan disease, or an infectious disease. In another embodiment, the subject is a mammal (e.g., a human) at risk of developing a genetic disease, an orphan disease, or an infectious disease.

[0071] As used herein, the term "administering" refers to providing or giving a pharmaceutical agent to a subject by any effective route. Exemplary routes of administration include, but are not limited to, one or more of the following: injection (e.g., subcutaneous, intramuscular, intradermal, intraperitoneal, intrathecal, intracerebroventricular, or intravenous), oral, intra-biliary duct, sublingual, rectal, transdermal, intranasal, vaginal, and inhalation. When used to treat a disease, disorder, condition, or symptom thereof, administration of a substance is typically carried out after the onset of the disease, disorder, condition, or symptom. When used to prevent a disease, disorder, condition, or symptom, administration of a substance is typically carried out prior to the onset of the disease, disorder, condition, or symptom.

[0072] As used herein, the term "microRNA" or "miRNA" refers to a non-coding small RNA. Generally, microRNAs are numbered according to the order in which they are discovered, and microRNAs of the same number refer to microRNAs that originate from the same pre-miRNA, for example, miRNA-142-3p refers to the miRNA-142 mature body that originates from the 3' arm of pre-miRNA-142, and miRNA-142-5p refers to the miRNA-142 mature body that originates from the 5' arm of pre-miRNA-142. As used herein, unless specifically indicated, if a microRNA is not indicated as "5p" or "3p", it refers to "5p" and / or "3p". For example, unless specifically indicated, miR-142 refers to miRNA-142-5p and / or miRNA-142-3p.

[0073] As used herein, the term "microRNA binding site" refers to a polynucleotide, such as DNA or RNA, that has sufficient complementarity to all or a portion of a microRNA to enable it to interact with, associate with, or bind to the microRNA.

[0074] Herein, some elements of the application will be described. These elements and specific embodiments are presented together, however it should be understood that they can be combined in any manner and in any number to produce additional embodiments. The different described examples and preferred embodiments should not be interpreted as limiting the application to only the explicitly described embodiments. The specification should be understood as supporting and including embodiments that combine the explicitly described embodiments with any number of the disclosed and / or preferred elements. Furthermore, unless the context indicates otherwise, any permutation and combination of all described elements in the application should be considered as being disclosed by the specification of the application. For example, in one embodiment, the polymeric lipid is PEG-DMG; in another embodiment, the LNPs comprise a polymeric lipid that is present in the total lipid present in the LNPs at 0.201 mol% to 0.595 mol%, then the following scenario is also an embodiment for which the application claims protection: the LNPs comprise DMG-PEG that is present in the total lipid present in the LNPs at 0.201 mol% to 0.595 mol%.

[0075] II. Lipid Nanoparticles (LNPs)

[0076] Current lipid nanoparticles (LNPs) mainly accumulate in the liver after entering the body, so that the active ingredients (e.g., mRNA) encapsulated by the LNPs are mainly delivered to the liver. However, for immune-related diseases such as inflammation, autoimmune diseases, infections and cancers, the active ingredients encapsulated by the LNPs are expected to be delivered to the spleen or immune cells, rather than the liver. Therefore, there is a need for LNPs that can target the delivery of encapsulated active ingredients to the spleen or immune cells. The inventors of the present application unexpectedly found that adjusting (reducing) the content of polymeric lipids in LNPs can change the delivery specificity of the LNPs, target the LNPs to the spleen or immune cells, and mainly deliver the active ingredients (e.g., mRNA) encapsulated by the LNPs to the spleen or immune cells, thereby increasing the amount of active ingredients delivered to the spleen or immune cells, and increasing the spleen / liver delivery ratio of the active ingredients. The spleen / liver delivery ratio of the active ingredients refers to the ratio of the amount of active ingredients delivered to the spleen to the amount of active ingredients delivered to the liver.

[0077] The present disclosure provides a lipid nanoparticle (LNP) comprising an ionizable lipid, a helper lipid, a structural lipid, and a polymeric lipid, the polymeric lipid accounting for 0 mol% to 0.8 mol% of the total lipid present in the LNP.

[0078] Polymeric lipid refers to a lipid comprising repeating subunits in the chemical structure. In some embodiments, the polymeric lipid is a lipid comprising a polymeric component.

[0079] In some embodiments, the lipid used to form the polymeric lipid comprises one or more of: 1,2-dimyristoyl-sn-glycerol (DMG), distearoyl-phosphatidyl-ethanolamine (DSPE), diacylglycerol (DAG), dialkyloxypropyl (DAA), phospholipid, ceramide (Cer), 1,2-distearoyl-rac-glycerol (DSG), dipalmitoyl, dioleoyl, distearoyl, dipalmitoyl phosphatidyl ethanolamine (DPPE), 1,2-dimyristyl acylpropyl-3-amine (DMA), phosphatidyl ethanolamine (PE), DOMG, disuccinic acid (DSA), 1,2-dinapthoyl-sn-glycero-3-phosphoethanolamine (DLPE), 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine (DMPE), PEG-1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), and dipalmitoyl-rac-glycero (DPG).

[0080] In some embodiments, the polymer used to form the polymeric lipid comprises one or both of: a hydrophilic polymer and an amphiphilic polymer.

[0081] In some embodiments, the polymer used to form the polymeric lipid is a hydrophilic polymer. In other embodiments, the polymer used to form the polymeric lipid is an amphiphilic polymer.

[0082] In some embodiments, the hydrophilic polymer comprises one or more of: polyethylene glycol (PEG), poly(oxazolines) (POX), poly(glycerols) (PGs), poly(hydroxypropyl methacrylate) (PHPMA), poly(2-hydroxyethyl methacrylate) (PHEMA), poly(N-(2-hydroxypropyl)methacrylamide) (HPMA), poly(vinylpyrrolidone) (PVP), poly(N,N-dimethyl acrylamide) (PDMA), poly(N-acryloyl morpholine) (PAcM), polyaminoacids, glycosaminoglycans (GAGs), heparin, hyaluronic acid (HA), polysialic acid (PSA), elastin-like polypeptides (ELPs), serum albumin, and CD47. In one alternative specific example, the hydrophilic polymer is polyethylene glycol (PEG).

[0083] In some embodiments, the polymeric lipid in the LNPs described above comprises one or more of: a polyethylene glycol-lipid (PEG-lipid), a poly(oxazoline)-lipid, a poly(glycerol)-lipid, a poly(hydroxypropyl methacrylate)-lipid, a poly(2-hydroxyethyl methacrylate)-lipid, a poly(N-(2-hydroxypropyl)methacrylamide)-lipid, a poly(vinylpyrrolidone)-lipid, a poly(N,N-dimethyl acrylamide)-lipid, a poly(N-acryloyl morpholine)-lipid, a glycosaminoglycan-lipid, a heparin-lipid, a hyaluronic acid-lipid, a polysialic acid-lipid, an elastin-like polypeptide-lipid, a serum albumin-lipid, and a CD47-lipid. Note that a “PEG-lipid” is a conjugate comprising polyethylene glycol and a lipid, a “poly(oxazoline)-lipid” refers to a conjugate comprising a poly(oxazoline) and a lipid, a “poly(glycerol)-lipid” refers to a conjugate comprising a poly(glycerol) and a lipid, and so on for the other polymeric lipids.

[0084] In some embodiments, the polymeric lipid of the LNP described above is a PEG-lipid.

[0085] In some embodiments, the polymeric lipid of the LNP described above is a PEG-lipid.

[0086] In some embodiments, the PEG lipid comprises one or more of: 1,2-dimyristoyl-sn- glyceromethoxypolyethyleneglycol (PEG-DMG or DMG-PEG), 1,2-distearoyl-sn-glycero-3- phosphoethanolamine-n-[amino(polyethyleneglycol)] (PEG-DSPE), PEG-diacetylglycerol (PEG- DSG or DSG-PEG), PEG-dipalmitoyl, PEG-dioleoyl, PEG-distearoyl, PEG-diacylglycerol (PEG- DAG), PEG-dipalmitoyl phosphatidylethanolamine (PEG-DPPE), PEG-1,2-dimyristyl acryloyl- 3-amine (PEG-c-DMA), dialkyloxypropyl-PEG (PEG-DAA), PEG-phospholipid, PEG- ceramide (PEG-Cer), PEG-1,2-dipalmitoyl-rac-glycerol (PEG-DPG or DPG-PEG), PEG- phosphatidylethanolamine (PEG-PE), PEG-c-DOMG, PEG-C-DSA, PEG-1,2-dinapthoyl-sn- glycero-3-phosphoethanolamine (PEG-DLPE), PEG-1,2-dimyristoyl-sn-glycero-3- phosphoethanolamine (PEG-DMPE), and PEG-1,2-dipalmitoyl-sn-glycero-3-phosphocholine (PEG- DPPC).

[0087] In some embodiments, the PEG lipid is one or more of: DMG-PEG, DSG-PEG, and DPG-PEG.

[0088] In some embodiments, the average molecular weight of the PEG in the PEG lipid is about 2000-5000. In an alternative specific example, the average molecular weight of the PEG in the PEG lipid is about 2000. In an alternative specific example, the polymeric lipid is DMG-PEG 2000.

[0089] In some embodiments, the amphiphilic polymer comprises one or more of: poly(carboxybetaine) (pCB), poly(sulfobetaine) (pSB), phosphobetaine-base polymers, and phosphorylcholine polymers. In some embodiments, the amphiphilic polymer comprises one or more of: poly(carboxybetaine acrylamide) (pCBAA), poly(carboxybetaine methacrylate), poly(sulfobetaine methacrylate), poly(methacryloyloxyethyl phosphorylcholine), poly(vinyl-pyridinio propanesulfonate), poly(carboxybetaine) based on vinylimidazole, poly(sulfobetaine) based on vinylimidazole, poly(sulfobetaine) based on vinylpyridine.

[0090] In some embodiments, the polymeric lipid comprises one or more of: polyhydroxybetaine-lipid, poly(sulfobetaine)-lipid, phosphobetaine-base polymer-lipid, and phosphorylcholine polymer-lipid. In some embodiments, the polymeric lipid comprises one or more of: poly(carboxybetaine acrylamide)-lipid, poly(carboxybetaine methacrylate)-lipid, poly(sulfobetaine methacrylate)-lipid, poly(methacryloyloxyethyl phosphorylcholine)-lipid, poly(vinyl-pyridinio propanesulfonate)-lipid, poly(carboxybetaine) based on vinylimidazole-lipid, poly(sulfobetaine) based on vinylimidazole-lipid, poly(sulfobetaine) based on vinylpyridine-lipid.

[0091] In some embodiments, the PEG-lipid is a compound of Formula (IV), or a salt thereof, or a stereoisomer thereof:

[0092] wherein:

[0093] R 3 is OR 0 ;

[0094] R 0 is hydrogen, optionally substituted alkyl, or an oxygen protecting group;

[0095] g is an integer between 1 and 100;

[0096] L1is optionally substituted C1-C 10 alkylene, wherein at least one methylene of the optionally substituted C1-C 10 alkylene is independently replaced with optionally substituted carbocyclylene, optionally substituted heterocyclylene, optionally substituted arylene, optionally substituted heteroarylene, -O-, -N(R M )-, -S-, -C(O)-, -C(O)N(R M )-, -NR M C(O)-, -C(O)O-, -OC(O)-, -OC(O)O-, -OC(O)N(R M )-, -NR M C(O)O-, or -NR M C(O)N(R M )-;

[0097] D is a click chemistry derived group or a group cleavable under physiological conditions;

[0098] h is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;

[0099] A is

[0100] each L2is independently a bond or optionally substituted C1-C6alkylene; optionally one methylene of the optionally substituted C1-C6alkylene is replaced with -O-, -N(R M )-, -S-, -C(O)-, -C(O)N(R M )-, -NR M C(O)-, -C(O)O-, -OC(O)-, -OC(O)O-, -OC(O)N(R M )-, -NR M C(O)O-, or -NR M C(O)N(R M )-;

[0101] each R 2 is independently optionally substituted C1-C 30 alkyl, optionally substituted C1-C 30 alkenyl, or optionally substituted C1-C 30alkynyl; optionally, R 2 one or more methylenes are independently replaced with an optionally substituted carbocyclylene, an optionally substituted heterocyclylene, an optionally substituted arylene, an optionally substituted heteroarylene, -N(R M )-, -O-, -S-, -C(O)-, -C(O)N(R M )-, -NR M C(O)-, -NR M C(O)N(R M )-, -C(O)O-, -OC(O)-, -OC(O)O-, -OC(O)N(R M )-, -NR M C(O)O-, -C(O)S-, -SC(O)-, -C(=NR M )-, -C(=NR M )N(R M )-, -NR M C(=NR M )-, -NR M C(=NR M )N(R M )-, -C(S)-, -C(S)N(R M )-, -NR M C(S)-, -NR M C(S)-, -NR M C(S)N(R M )-, -S(O)-, -OS(O)-, -S(O)O-, -OS(O)O-, -OS(O)2-, -S(O)2O-, -OS(O)2O-, -N(R M )S(O)-, S(O)N(R M )-, -N(R M )S(O)N(R M )-, -OS(O)N(R M )-, -N(R M )S(O)O-, -S(O)2-, N(R M )S(O)2-, -S(O)2N(R M )-, -N(R M )S(O)2N(R M )-, -OS(O)2N(R M )-, or -N(R M )S(O)2O-;

[0102] each R M is independently hydrogen, optionally substituted alkyl, or a nitrogen protecting group;

[0103] Ring B is an optionally substituted carbocyclic, optionally substituted heterocyclic, optionally substituted aryl, or optionally substituted heteroaryl; and

[0104] k is equal to 1 or 2.

[0105] In some embodiments, the compound of Formula (IV) is a PEG-OH lipid (i.e., R 0 is H).

[0106] In some embodiments, the PEG lipid is a compound of Formula (V), or a salt thereof, or a stereoisomer thereof:

[0107] wherein:

[0108] R 3 is OR 0 ;

[0109] R 0 is hydrogen, optionally substituted alkyl, or an oxygen protecting group;

[0110] g is an integer between 1 and 100;

[0111] R 5 is optionally substituted C 10 -C 40 alkyl, optionally substituted C 10 -C 40 alkenyl, or optionally substituted C 10 -C 40 alkynyl; optionally, one or more methylene groups of R 5 are independently replaced by optionally substituted carbocyclyl ene, optionally substituted heterocyclyl ene, optionally substituted arylene, optionally substituted heteroarylene, -N(R M )-, -O-, -S-, -C(O)-, -C(O)N(R M )-, -NR M C(O)-, -NR M C(O)N(R M )-, -C(O)O-, -OC(O)-, -OC(O)O-, -OC(O)N(R M )-, -NR M C(O)O-, -C(O)S-, -SC(O)-, -C(=NR M )-, -C(=NR M )N(R M )-, -NR M C(=NR M )-, -NR M C(=NR M )N(R M )-, -C(S)-, -C(S)N(RM )-, -NR M C(S)-, -NR M C(S)-, -NR M C(S)N(R M )-, -S(O)-, -OS(O)-, -S(O)O-, -OS(O)O-, -OS(O)2-, -S(O)2O-, -OS(O)2O-, -N(R M )S(O)-, S(O)N(R M )-, -N(R M )S(O)N(R M )-, -OS(O)N(R M )-, -N(R M )S(O)O-, -S(O)2-, N(R M )S(O)2-, -S(O)2N(R M )-, -N(R M )S(O)2N(R M )-, -OS(O)2N(R M )- or -N(R M )S(O)2O- in place of the -C(O)- group.

[0112] each R M is independently hydrogen, optionally substituted alkyl, or a nitrogen protecting group.

[0113] In some embodiments, the compound of Formula (V) is a PEG-OH lipid (i.e., R 0 is H).

[0114] Further, in some embodiments, the polymeric lipid can also be any one or more of the PEG lipids in Hoang Thi, Thai Thanh, et al. Polymers, 12(2), 298 (2020), the PEG lipids in the above- referenced document being incorporated herein by reference.

[0115] The present disclosure provides a lipid nanoparticle (LNP) comprising an ionizable lipid, a helper lipid, a structural lipid, and a PEG lipid, the PEG lipid being present at 0 mol% to 0.8 mol% of the total lipid present in the LNP.

[0116] The present disclosure provides a lipid nanoparticle (LNPs) comprising an ionizable lipid, a helper lipid, a structural lipid, and a polymer lipid, the polymer lipid being present at 0 mol% to 0.8 mol% of the total lipid present in the LNPs, the ionizable lipid being present at 30 mol% to 60 mol% of the total lipid present in the LNPs, the helper lipid being present at 0 mol% to 30 mol% of the total lipid present in the LNPs, and the structural lipid being present at 10 mol% to 60 mol% of the total lipid present in the LNPs.

[0117] The present disclosure provides a lipid nanoparticle (LNPs) comprising an ionizable lipid, a helper lipid, a structural lipid, and a PEG lipid, the PEG lipid being present at 0 mol% to 0.8 mol% of the total lipid present in the LNPs, the ionizable lipid being present at 30 mol% to 60 mol% of the total lipid present in the LNPs, the helper lipid being present at 0 mol% to 30 mol% of the total lipid present in the LNPs, and the structural lipid being present at 10 mol% to 60 mol% of the total lipid present in the LNPs.

[0118] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0001 mol% to 0.8 mol% of the total lipid present in the LNP. For example, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0001 mol%, 0.00011 mol%, 0.0002 mol%, 0.0003 mol%, 0.0004 mol%, 0.00045 mol%, 0.00049 mol%, 0.0005 mol%, 0.00051 mol%, 0.00055 mol%, 0.0006 mol%, 0.00065 mol%, 0.0007 mol%, 0.00075 mol%, 0.0008 mol%, 0.00085 mol%, 0.0009 mol%, 0.00095 mol%, 0.00099 mol%, 0.001 mol%, 0.0011 mol%, 0.0015 mol%, 0.002 mol%, 0.0025 mol%, 0.003 mol%, 0.0035 mol%, 0.004 mol%, 0.0045 mol%, 0.0049 mol%, 0.005 mol%, 0.0051 mol%, 0.0055 mol%, 0.006 mol%, 0.0065 mol%, 0.007 mol%, 0.0075 mol%, 0.008 mol%, 0.0085 mol%, 0.009 mol%, 0.0095 mol%, 0.0099 mol%, 0.01 mol%, 0.011 mol%, 0.015 mol%, 0.02 mol%, 0.025 mol%, 0.03 mol%, 0.035 mol%, 0.04 mol%, 0.045 mol%, 0.049 mol%, 0.05 mol%, 0.051 mol%, 0.051 mol%, 0.055 mol%, 0.06 mol%, 0.065 mol%, 0.07 mol%, 0.075 mol%, 0.08 mol%, 0.085 mol%, 0.09 mol%, 0.095 mol%, 0.0999 mol%, 0.1 mol%, 0.101 mol%, 0.105 mol%, 0.11 mol%, 0.115 mol%, 0.12 mol%, 0.125 mol%, 0.13 mol%, 0.135 mol%, 0.14 mol%, 0.145 mol%, 0.149 mol%, 0.15 mol%, 0.151 mol%, 0.155 mol%, 0.16 mol%, 0.165 mol%, 0.17 mol%, 0.175 mol%, 0.18 mol%, 0.185 mol%, 0.19 mol%, 0.195 mol%, 0.0.2 mol %, 0.201 mol %, 0.205 mol %, 0.21 mol %, 0.215 mol %, 0.22 mol %, 0.225 mol %, 0.23 mol %, 0.235 mol %, 0.24 mol %, 0.245 mol %, 0.249 mol %, 0.25 mol %, 0.251 mol %, 0.255 mol %, 0.26 mol %, 0.265 mol %, 0.27 mol %, 0.275 mol %, 0.28 mol %, 0.285 mol %, 0.29 mol %, 0.295 mol %, 0.299 mol %, 0.3 mol %, 0.301 mol %, 0.305 mol %, 0.31 mol %, 0.315 mol %, 0.32 mol %, 0.325 mol %, 0.33 mol %, 0.335 mol %, 0.34 mol %, 0.345 mol %, 0.349 mol %, 0.35 mol %, 0.351 mol %, 0.355 mol %, 0.36 mol %, 0.365 mol %, 0.37 mol %, 0.375 mol %, 0.38 mol %, 0.385 mol %, 0.39 mol %, 0.395 mol %, 0.395 mol %, 0.399 mol %, 0.4 mol %, 0.401 mol %, 0.405 mol %, 0.41 mol %, 0.415 mol %, 0.42 mol %, 0.425 mol %, 0.43 mol %, 0.435 mol %, 0.44 mol %, 0.445 mol %, 0.449 mol %, 0.45 mol %, 0.451 mol %, 0.455 mol %, 0.46 mol %, 0.465 mol %, 0.47 mol %, 0.475 mol %, 0.48 mol %, 0.485 mol %, 0.49 mol %, 0.495 mol %, 0.499 mol %, 0.5 mol %, 0.501 mol %, 0.505 mol %, 0.51 mol %, 0.515 mol %, 0.52 mol %, 0.525 mol %, 0.53 mol %, 0.535 mol %, 0.54 mol %, 0.545 mol %, 0.549 mol %, 0.55 mol %, 0.551 mol %, 0.555 mol %, 0.56 mol %, 0.565 mol %, 0.57 mol %, 0.575 mol %, 0.58 mol %, 0.585 mol %, 0.59 mol %, 0.595 mol %, 0.599 mol %, 0.6 mol %, 0.601 mol %, 0.605 mol %, 0.61 mol %, 0.615 mol %, 0.62 mol %, 0.625 mol%, 0.63 mol%, 0.635 mol%, 0.64 mol%, 0.645 mol%, 0.649 mol%, 0.65 mol%, 0.651 mol%, 0.655 mol%, 0.66 mol%, 0.665 mol%, 0.67 mol%, 0.675 mol%, 0.68 mol%, 0.685 mol%, 0.69 mol%, 0.695 mol%, 0.699 mol%, 0.7 mol%, 0.701 mol%, 0.705 mol%, 0.71 mol%, 0.715 mol%, 0.72 mol%, 0.725 mol%, 0.73 mol%, 0.735 mol%, 0.74 mol%, 0.745 mol%, 0.749 mol%, 0.75 mol%, 0.751 mol%, 0.755 mol%, 0.76 mol%, 0.765 mol%, 0.77 mol%, 0.775 mol%, 0.78 mol%, 0.785 mol%, or 0.79 mol%.

[0119] In some embodiments, the polymer lipids (e.g., PEG lipids) in the LNPs comprise 0.0001 mol% to 0.79 mol%, 0.0001 mol% to 0.75 mol%, 0.0001 mol% to 0.7 mol%, 0.0001 mol% to 0.65 mol%, 0.0001 mol% to 0.6 mol%, 0.0001 mol% to 0.55 mol%, 0.0001 mol% to 0.5 mol%, 0.0001 mol% to 0.45 mol%, 0.0001 mol% to 0.4 mol%, 0.0001 mol% to 0.35 mol%, and 0.0001 mol% to 0.0001 mol% of the total lipids present in the LNPs. %~0.3mol%, 0.0001mol%~0.25mol%, 0.0001mol%~0.2mol%, 0.0001mol%~0.15mol%, 0.0001mol%~0.1mol%, 0.0001mol%~0.05mol%, 0.0001mol%~0.01m ol%, 0.0001mol%~0.005mol%, 0.0001mol%~0.001mol%, 0.0001mol%~0.0005mol%, 0.0005mol%~0.79mol%, 0.0005mol%~0.75mol%, 0.0005mol%~0.7mol %, 0.0005mol%~0.65mol%, 0.0005mol%~0.6mol%, 0.0005mol%~0.55mol%, 0.0005mol%~0.5mol%, 0.0005mol%~0.45mol%, 0.0005mol%~0.4mol%, 0.000 5mol%~0.35mol%, 0.0005mol%~0.3mol%, 0.0005mol%~0.25mol%, 0.0005mol%~0.2mol%, 0.0005mol%~0.15mol%, 0.0005mol%~0.1mol%, 0.0005mol%~0 .05mol%, 0.0005mol%~0.01mol%, 0.0005mol%~0.005mol%, 0.0005mol%~0.001mol%, 0.001mol%~0.79mol%, 0.001mol%~0.75mol%, 0.001mol%~0.7mol %, 0.001mol%~0.65mol%, 0.001mol%~0.6mol%, 0.001mol%~0.55mol%, 0.001mol%~0.5mol%, 0.001mol%~0.45mol%, 0.001mol%~0.4mol%, 0.001mol%~0.35 mol%, 0.001 mol%~0.3 mol%, 0.001 mol%~0.25 mol%, 0.001 mol%~0.2 mol%, 0.001 mol%~0.15 mol%, 0.001 mol%~0.1 mol%, 0.001 mol%~0.05 mol%, 0.001 m ol%~0.01mol%、0.001mol%~0.005mol%、0.005mol%~0.79mol%、0.005mol%~ 0.75mol%、0.005mol%~0.7mol%、0.005mol%~0.65mol%、0.005mol%~0.6mol %、0.005mol%~0.55mol%、0.005mol%~0.5mol%、0.005mol%~0.45mol%、0.005mol%~0.4mol%、0.005mol%~0.35mol%、0.005mol%~0.3mol%、0.005mol%~0 .25mol%、0.005mol%~0.2mol%、0.005mol%~0.15mol%、0.005mol%~0.1mol% 、0.005mol%~0.05mol%、0.005mol%~0.01mol%、0.01mol%~0.79mol%、0.01m ol%~0.75mol%、0.01mol%~0.7mol%、0.01mol%~0.65mol%、0.01mol%~0.6mo l%、0.01mol%~0.55mol%、0.01mol%~0.5mol%、0.01mol%~0.45mol%、0.01mo l%~0.4mol%、0.01mol%~0.35mol%、0.01mol%~0.3mol%、0.01mol%~0.25mol %、0.01mol%~0.2mol%、0.01mol%~0.15mol%、0.01mol%~0.1mol%、0.01mol% ~0.05mol%、0.05mol%~0.79mol%、0.05mol%~0.75mol%、0.05mol%~0.7mol% 、0.05mol%~0.65mol%、0.05mol%~0.6mol%、0.05mol%~0.55mol%、0.05mol% ~0.5mol%、0.05mol%~0.45mol%、0.05mol%~0.4mol%、0.05mol%~0.35mol%、 0.05mol%~0.3mol%、0.05mol%~0.25mol%、0.05mol%~0.2mol%、0.05mol%~0.15 mol%, 0.05 mol% to 0.1 mol%, 0.1 mol% to 0.79 mol%, 0.1 mol% to 0.75 mol%, 0.1 mol% to 0.7 mol%, 0.1 mol% to 0.65 mol%, 0.1 mol% to 0.6 mol%, 0.1 mol% to 0.55 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.45 mol%, 0.1 mol% to 0.4 mol%, 0.1 mol% to 0.35 mol%, 0.1 mol% to 0.3 mol%, 0.1 mol% to 0.25 mol%, 0.1 mol% to 0.2 mol%, 0.1 mol% to 0.15 mol%, 0.15 mol% to 0.79 mol%, 0.15 mol% to 0.75 mol%, 0.15 mol% to 0.7 mol%, 0.15 mol% to 0.65 mol%, 0.15 mol% to 0.6 mol%, 0.15 mol% to 0.55 mol%, 0.15 mol% to 0.5 mol%, 0.15 mol% to 0.45 mol%, 0.15 mol% to 0.4 mol%, 0.15 mol% to 0.35 mol%, 0.15 mol% to 0.3 mol%, 0.15 mol% to 0.25 mol%, 0.15 mol% to 0.2 mol%, 0.2 mol% to 0.79 mol%, 0.2 mol% to 0.75 mol%, 0.2 mol% to 0.7 mol%, 0.2 mol% to 0.65 mol%, 0.2 mol% to 0.6 mol%, 0.2 mol% to 0.55 mol%, 0.2 mol% to 0.5 mol%, 0.2 mol% to 0.45 mol%, 0.2 mol% to 0.4 mol%, 0.2 mol% to 0.35 mol%, 0.2 mol% to 0.3 mol%, 0.2 mol% to 0.25 mol%, 0.3 mol% to 0.79 mol%, 0.3 mol% to 0.75 mol%, 0.3 mol% to 0.7 mol%, 0.3 mol% to 0.65 mol%, 0.3 mol% to 0.6 mol%, 0.3 mol% to 0.55 mol%, 0.3 mol% to 0.5 mol%, 0.3 mol% to 0.45 mol%, 0.3 mol% to 0.4 mol%, 0.3 mol% to 0.35 mol%, 0.4 mol% to 0.79 mol%, 0.4 mol% to 0.75 mol%, 0.4 mol% to 0.7 mol%, 0.4 mol% to 0.65 mol%, 0.4 mol% to 0.6 mol%, 0.4 mol% to 0.55 mol%, 0.4 mol% to 0.5 mol%, 0.4 mol% to 0.45 mol%, 0.5 mol% to 0.79 mol%, 0.5 mol% to 0.75 mol%, 0.5 mol% to 0.7 mol%, 0.5 mol% to 0.65 mol%, 0.5 mol% to 0.6 mol%, 0.5 mol% to 0.55 mol%, 0.6 mol% to 0.79 mol%, 0.6 mol% to 0.75 mol%, 0.6 mol% to 0.7 mol%, 0.6 mol% to 0.65 mol%, 0.7 mol% to 0.79 mol%, or 0.7 mol% to 0.75 mol%.

[0120] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNPs is present at 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.7 mol%, 0.01 mol% to 0.6 mol%, 0.01 mol% to 0.5 mol%, 0.01 mol% to 0.4 mol%, 0.01 mol% to 0.3 mol%, 0.01 mol% to 0.2 mol%, 0.01 mol% to 0.1 mol%, 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.7 mol%, 0.05 mol% to 0.6 mol%, 0.05 mol% to 0.5 mol%, 0.05 mol% to 0.4 mol%, 0.05 mol% to 0.3 mol%, 0.05 mol% to 0.25 mol%, 0.05 mol% to 0.2 mol%, 0.05 mol% to 0.15 mol%, 0.05 mol% to 0.1 mol%, 0.1 mol% to 0.8 mol%, 0.1 mol% to 0.7 mol%, 0.1 mol% to 0.6 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.4 mol%, 0.1 mol% to 0.3 mol%, 0.1 mol% to 0.2 mol%, 0.1 mol% to 0.15 mol%, 0.15 mol% to 0.8 mol%, 0.15 mol% to 0.7 mol%, 0.15 mol% to 0.6 mol%, 0.15 mol% to 0.5 mol%, 0.15 mol% to 0.4 mol%, 0.15 mol% to 0.3 mol%, 0.15 mol% to 0.25 mol%, 0.2 mol% to 0.8 mol%, 0.2 mol% to 0.7 mol%, 0.2 mol% to 0.6 mol%, 0.2 mol% to 0.5 mol%, 0.2 mol% to 0.4 mol%, or 0.2 mol% to 0.35 mol% of the total lipid present in the LNP.

[0121] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above- described LNPs is present at 0.0001 mol% to 0.8 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above- described LNPs is present at 0.005 mol% to 0.8 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above- described LNPs is present at 0.01 mol% to 0.8 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above- described LNPs is present at 0.05 mol% to 0.8 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above- described LNPs is present at 0.1 mol% to 0.8 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above- described LNPs is present at 0.2 mol% to 0.8 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above- described LNPs is present at 0.3 mol% to 0.8 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above- described LNPs is present at 0.4 mol% to 0.8 mol% of the total lipid present in the LNP.

[0122] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.0005 mol% to 0.7 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.001 mol% to 0.7 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.005 mol% to 0.7 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.01 mol% to 0.7 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.05 mol% to 0.7 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.1 mol% to 0.7 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.2 mol% to 0.7 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.3 mol% to 0.7 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.4 mol% to 0.7 mol% of the total lipid present in the LNP.

[0123] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.0005 mol% to 0.65 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.001 mol% to 0.65 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.005 mol% to 0.65 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.01 mol% to 0.65 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.05 mol% to 0.65 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.1 mol% to 0.65 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.2 mol% to 0.65 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.3 mol% to 0.65 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.4 mol% to 0.65 mol% of the total lipid present in the LNP.

[0124] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0001 mol% to 0.6 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0005 mol% to 0.6 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.001 mol% to 0.6 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.005 mol% to 0.6 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.01 mol% to 0.6 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.05 mol% to 0.6 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.1 mol% to 0.6 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.2 mol% to 0.6 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.3 mol% to 0.6 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.4 mol% to 0.6 mol% of the total lipid present in the LNP.

[0125] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0001 mol% to 0.5 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0005 mol% to 0.5 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.001 mol% to 0.5 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.005 mol% to 0.5 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.01 mol% to 0.5 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.05 mol% to 0.5 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.06 mol% to 0.5 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.07 mol% to 0.5 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.08 mol% to 0.5 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.09 mol% to 0.5 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.1 mol% to 0.5 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.2 mol% to 0.5 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.3 mol% to 0.5 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.4 mol% to 0.5 mol% of the total lipid present in the LNP.

[0126] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.0001 mol% to 0.4 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.0005 mol% to 0.4 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.001 mol% to 0.4 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.005 mol% to 0.4 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.01 mol% to 0.4 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.05 mol% to 0.4 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.06 mol% to 0.4 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.07 mol% to 0.4 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.08 mol% to 0.4 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.09 mol% to 0.4 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.1 mol% to 0.4 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.2 mol% to 0.4 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.3 mol% to 0.4 mol% of the total lipid present in the LNP.

[0127] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.0001 mol% to 0.35 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.0005 mol% to 0.35 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.001 mol% to 0.35 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.005 mol% to 0.35 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.01 mol% to 0.35 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.05 mol% to 0.35 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.1 mol% to 0.35 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.2 mol% to 0.35 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.3 mol% to 0.35 mol% of the total lipid present in the LNP.

[0128] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.0001 mol% to 0.3 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.0005 mol% to 0.3 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.001 mol% to 0.3 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.005 mol% to 0.3 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.01 mol% to 0.3 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.05 mol% to 0.3 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.06 mol% to 0.3 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.07 mol% to 0.3 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.08 mol% to 0.3 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.09 mol% to 0.3 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.1 mol% to 0.3 mol% of the total lipid present in the LNP. The polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.15 mol% to 0.3 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.2 mol% to 0.3 mol% of the total lipid present in the LNP.

[0129] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.0001 mol% to 0.25 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.0005 mol% to 0.25 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.001 mol% to 0.25 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.005 mol% to 0.25 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.01 mol% to 0.25 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.05 mol% to 0.25 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.1 mol% to 0.25 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.15 mol% to 0.25 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.2 mol% to 0.25 mol% of the total lipid present in the LNP.

[0130] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above- described LNPs is present at 0.0001 mol% to 0.2 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above- described LNPs is present at 0.0005 mol% to 0.2 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above- described LNPs is present at 0.001 mol% to 0.2 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above- described LNPs is present at 0.005 mol% to 0.2 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above- described LNPs is present at 0.01 mol% to 0.2 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above- described LNPs is present at 0.05 mol% to 0.2 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above- described LNPs is present at 0.06 mol% to 0.2 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above- described LNPs is present at 0.07 mol% to 0.2 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above- described LNPs is present at 0.08 mol% to 0.2 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above- described LNPs is present at 0.09 mol% to 0.2 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above- described LNPs is present at 0.1 mol% to 0.2 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above- described LNPs is present at 0.15 mol% to 0.2 mol% of the total lipid present in the LNP.

[0131] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.0001 mol% to 0.15 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.0005 mol% to 0.15 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.001 mol% to 0.15 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.005 mol% to 0.15 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.01 mol% to 0.15 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.05 mol% to 0.15 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.06 mol% to 0.15 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.07 mol% to 0.15 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.08 mol% to 0.15 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.09 mol% to 0.15 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.1 mol% to 0.15 mol% of the total lipid present in the LNP.

[0132] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.06 mol% to 0.1 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.07 mol% to 0.1 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.08 mol% to 0.1 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.09 mol% to 0.1 mol% of the total lipid present in the LNP.

[0133] In some embodiments, the proportion of the polymeric lipid (e.g., PEG lipid) in the above-described LNPs to the total lipid present in the LNP is suitably not one or more of: 0 mol%, 0.0001 mol%, 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.55 mol%, 0.6 mol%, 0.65 mol%, 0.7 mol%, 0.75 mol%, and 0.8 mol%. For example, in some embodiments, the proportion of the polymeric lipid (e.g., PEG lipid) in the above-described LNPs to the total lipid present in the LNP is from 0 mol% to 0.8 mol% and is not 0 mol%, 0.0001 mol%, 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.55 mol%, 0.6 mol%, 0.65 mol%, 0.7 mol%, 0.75 mol%, and 0.8 mol%. For example, in some embodiments, the proportion of the polymeric lipid (e.g., PEG lipid) in the above-described LNPs to the total lipid present in the LNP is from 0.0001 mol% to 0.749 mol% and is not one or more of: 0.0001 mol%, 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.55 mol%, 0.6 mol%, 0.65 mol%, and 0.7 mol%.For example, in some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.0001 mol% to 0.5 mol% of the total lipid present in the LNPs and is not one or more of the following: 0.0001 mol%, 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, and 0.5 mol%. For example, in some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.0005 mol% to 0.4 mol% of the total lipid present in the LNPs and is not one or more of the following: 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, and 0.35 mol%.

[0134] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at a ratio of 0 mol% to 0.8 mol% of the total lipid present in the LNPs and is not one or more of the following: 0 mol%, 0.0001 mol%, 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.55 mol%, 0.6 mol%, 0.65 mol%, 0.7 mol%, 0.75 mol%, and 0.8 mol%.

[0135] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at a ratio of 0 mol% to 0.8 mol% of the total lipid present in the LNPs and is not one or more of the following: 0 mol%, 0.0001 mol%, 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, 0.7 mol%, 0.75 mol%, and 0.8 mol%.

[0136] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0005 mol% to 0.8 mol% of the total lipid present in the LNP and is not 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, 0.7 mol%, 0.75 mol%, and 0.8 mol%. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.001 mol% to 0.8 mol% of the total lipid present in the LNP and is not 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, 0.7 mol%, 0.75 mol%, and 0.8 mol%. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.005 mol% to 0.8 mol% of the total lipid present in the LNP and is not 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, 0.7 mol%, 0.75 mol%, and 0.8 mol%. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.01 mol% to 0.8 mol% of the total lipid present in the LNP and is not 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, 0.7 mol%, 0.75 mol%, and 0.8 mol%.In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.05 mol% to 0.8 mol% and not 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, 0.7 mol%, 0.75 mol%, and 0.8 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.1 mol% to 0.8 mol% and not 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, 0.7 mol%, 0.75 mol%, and 0.8 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.2 mol% to 0.8 mol% and not 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, 0.7 mol%, 0.75 mol%, and 0.8 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.3 mol% to 0.8 mol% and not 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, 0.7 mol%, 0.75 mol%, and 0.8 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.4 mol% to 0.8 mol% and not 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, 0.7 mol%, 0.75 mol%, and 0.8 mol% of the total lipid present in the LNP.

[0137] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0005 mol% to 0.7 mol% of the total lipid present in the LNP and is not 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, and 0.7 mol%. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.001 mol% to 0.7 mol% of the total lipid present in the LNP and is not 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, and 0.7 mol%. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.005 mol% to 0.7 mol% of the total lipid present in the LNP and is not 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, and 0.7 mol%. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.01 mol% to 0.7 mol% of the total lipid present in the LNP and is not 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, and 0.7 mol%. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.05 mol% to 0.7 mol% of the total lipid present in the LNP and is not 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, and 0.7 mol%.In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.1 mol% to 0.7 mol% and not 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, and 0.7 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.2 mol% to 0.7 mol% and not 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, and 0.7 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.3 mol% to 0.7 mol% and not 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, and 0.7 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.4 mol% to 0.7 mol% and not 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, and 0.7 mol% of the total lipid present in the LNP.

[0138] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0001 mol% to 0.6 mol% and not 0.0001 mol%, 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, and 0.6 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0005 mol% to 0.6 mol% and not 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, and 0.6 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.001 mol% to 0.6 mol% and not 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, and 0.6 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.005 mol% to 0.6 mol% and not 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, and 0.6 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.01 mol% to 0.6 mol% and not 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, and 0.6 mol% of the total lipid present in the LNP.In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.05 mol% to 0.6 mol% and not 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, and 0.6 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.1 mol% to 0.6 mol% and not 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, and 0.6 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.2 mol% to 0.6 mol% and not 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, and 0.6 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.3 mol% to 0.6 mol% and not 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, and 0.6 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.4 mol% to 0.6 mol% and not 0.4 mol%, 0.45 mol%, 0.5 mol%, and 0.6 mol% of the total lipid present in the LNP.

[0139] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0001 mol% to 0.5 mol% of the total lipid present in the LNP and is not 0.0001 mol%, 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, and 0.5 mol%. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0005 mol% to 0.5 mol% of the total lipid present in the LNP and is not 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, and 0.5 mol%. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.001 mol% to 0.5 mol% of the total lipid present in the LNP and is not 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, and 0.5 mol%. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.005 mol% to 0.5 mol% of the total lipid present in the LNP and is not 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, and 0.5 mol%. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.01 mol% to 0.5 mol% of the total lipid present in the LNP and is not 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, and 0.5 mol%.In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.05 mol% to 0.5 mol% and not 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, and 0.5 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.1 mol% to 0.5 mol% and not 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, and 0.5 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.2 mol% to 0.5 mol% and not 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, and 0.5 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.3 mol% to 0.5 mol% and not 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, and 0.5 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.4 mol% to 0.5 mol% and not 0.4 mol%, 0.45 mol%, and 0.5 mol% of the total lipid present in the LNP.

[0140] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0001 mol% to 0.45 mol% of the total lipid present in the LNP and is not 0.0001 mol%, 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, and 0.45 mol%. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0005 mol% to 0.45 mol% of the total lipid present in the LNP and is not 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, and 0.45 mol%. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.001 mol% to 0.45 mol% of the total lipid present in the LNP and is not 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, and 0.45 mol%. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.005 mol% to 0.45 mol% of the total lipid present in the LNP and is not 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, and 0.45 mol%. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.01 mol% to 0.45 mol% of the total lipid present in the LNP and is not 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, and 0.45 mol%.In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.05 mol% to 0.45 mol% and not 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, and 0.45 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.1 mol% to 0.45 mol% and not 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, and 0.45 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.2 mol% to 0.45 mol% and not 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, and 0.45 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.3 mol% to 0.45 mol% and not 0.3 mol%, 0.35 mol%, 0.4 mol%, and 0.45 mol% of the total lipid present in the LNP.

[0141] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0001 mol% to 0.4 mol% and not 0.0001 mol%, 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, and 0.4 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0005 mol% to 0.4 mol% and not 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, and 0.4 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.001 mol% to 0.4 mol% and not 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, and 0.4 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.005 mol% to 0.4 mol% and not 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, and 0.4 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.01 mol% to 0.4 mol% and not 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, and 0.4 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.05 mol% to 0.4 mol% and not 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, and 0.4 mol% of the total lipid present in the LNP.In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above- described LNPs is present at 0.1 mol% to 0.4 mol% and not 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, and 0.4 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.2 mol% to 0.4 mol% and not 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, and 0.4 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.3 mol% to 0.4 mol% and not 0.3 mol%, 0.35 mol%, and 0.4 mol% of the total lipid present in the LNP.

[0142] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0001 mol% to 0.35 mol% of the total lipid present in the LNP and is not 0.0001 mol%, 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, and 0.35 mol%. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0005 mol% to 0.35 mol% of the total lipid present in the LNP and is not 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, and 0.35 mol%. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.001 mol% to 0.35 mol% of the total lipid present in the LNP and is not 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, and 0.35 mol%. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.005 mol% to 0.35 mol% of the total lipid present in the LNP and is not 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, and 0.35 mol%. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.01 mol% to 0.35 mol% of the total lipid present in the LNP and is not 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, and 0.35 mol%. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.05 mol% to 0.35 mol% of the total lipid present in the LNP and is not 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, and 0.35 mol%.In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above- described LNPs is present at 0.1 mol% to 0.35 mol% and not 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, and 0.35 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.2 mol% to 0.35 mol% and not 0.2 mol%, 0.25 mol%, 0.3 mol%, and 0.35 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above-described LNPs is present at 0.3 mol% to 0.35 mol% and not 0.3 mol% and 0.35 mol% of the total lipid present in the LNP.

[0143] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0001 mol% to 0.3 mol% and not 0.0001 mol%, 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, and 0.3 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0005 mol% to 0.3 mol% and not 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, and 0.3 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.001 mol% to 0.3 mol% and not 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, and 0.3 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.005 mol% to 0.3 mol% and not 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, and 0.3 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.01 mol% to 0.3 mol% and not 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, and 0.3 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.05 mol% to 0.3 mol% and not 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, and 0.3 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.1 mol% to 0.3 mol% and not 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, and 0.3 mol% of the total lipid present in the LNP.In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.2 mol% to 0.3 mol% and not 0.2 mol%, 0.25 mol%, and 0.3 mol% of the total lipid present in the LNP.

[0144] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0001 mol% to 0.25 mol% and not 0.0001 mol%, 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, and 0.25 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0005 mol% to 0.25 mol% and not 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, and 0.25 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.001 mol% to 0.25 mol% and not 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, and 0.25 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.005 mol% to 0.25 mol% and not 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, and 0.25 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.01 mol% to 0.25 mol% and not 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, and 0.25 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.05 mol% to 0.25 mol% and not 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, and 0.25 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.1 mol% to 0.25 mol% and not 0.1 mol%, 0.15 mol%, 0.2 mol%, and 0.25 mol% of the total lipid present in the LNP.

[0145] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0001 mol% to 0.2 mol% and not 0.0001 mol%, 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, and 0.2 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0005 mol% to 0.2 mol% and not 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, and 0.2 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.001 mol% to 0.2 mol% and not 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, and 0.2 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.005 mol% to 0.2 mol% and not 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, and 0.2 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.01 mol% to 0.2 mol% and not 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, and 0.2 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.05 mol% to 0.2 mol% and not 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, and 0.2 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.1 mol% to 0.2 mol% and not 0.1 mol%, 0.15 mol%, and 0.2 mol% of the total lipid present in the LNP.

[0146] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0001 mol% to 0.15 mol% and not 0.0001 mol%, 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, and 0.15 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0005 mol% to 0.15 mol% and not 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, and 0.15 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.001 mol% to 0.15 mol% and not 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, and 0.15 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.005 mol% to 0.15 mol% and not 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, and 0.15 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.01 mol% to 0.15 mol% and not 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, and 0.15 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.05 mol% to 0.15 mol% and not 0.05 mol%, 0.07 mol%, 0.1 mol%, and 0.15 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.1 mol% to 0.15 mol% and not 0.1 mol% and 0.15 mol% of the total lipid present in the LNP.

[0147] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0001 mol% to 0.1 mol% and not 0.0001 mol%, 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, and 0.1 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0005 mol% to 0.1 mol% and not 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, and 0.1 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.001 mol% to 0.1 mol% and not 0.001 mol%, 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, and 0.1 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.005 mol% to 0.1 mol% and not 0.005 mol%, 0.01 mol%, 0.05 mol%, 0.07 mol%, and 0.1 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.01 mol% to 0.1 mol% and not 0.01 mol%, 0.05 mol%, 0.07 mol%, and 0.1 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.05 mol% to 0.1 mol% and not 0.05 mol%, 0.07 mol%, and 0.1 mol% of the total lipid present in the LNP.

[0148] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0001 mol% to 0.05 mol% and not 0.0001 mol%, 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, and 0.05 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0005 mol% to 0.05 mol% and not 0.0005 mol%, 0.001 mol%, 0.005 mol%, 0.01 mol%, and 0.05 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.001 mol% to 0.05 mol% and not 0.001 mol%, 0.005 mol%, 0.01 mol%, and 0.05 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.005 mol% to 0.05 mol% and not 0.005 mol%, 0.01 mol%, and 0.05 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.01 mol% to 0.05 mol% and not 0.01 mol% and 0.05 mol% of the total lipid present in the LNP.

[0149] In some embodiments, the proportion of the polymeric lipid (e.g., PEG lipid) in the above-described LNPs to the total lipid present in the LNP is: greater than 0 and less than 0.0001 mol%, greater than 0.0001 mol% and less than 0.0005 mol%, greater than 0.0005 mol% and less than 0.001 mol%, greater than 0.001 mol% and less than 0.005 mol%, greater than 0.005 mol% and less than 0.01 mol%, greater than 0.01 mol% and less than 0.05 mol%, greater than 0.05 mol% and less than 0.07 mol%, greater than 0.07 mol% and less than 0.1 mol%, greater than 0.1 mol% and less than 0.15 mol%, greater than 0.15 mol% and less than 0.2 mol%, greater than 0.2 mol% and less than 0.25 mol%, greater than 0.25 mol% and less than 0.3 mol%, greater than 0.3 mol% and less than 0.35 mol%, greater than 0.35 mol% and less than 0.4 mol%, greater than 0.4 mol% and less than 0.45 mol%, greater than 0.45 mol% and less than 0.5 mol%, greater than 0.5 mol% and less than 0.6 mol%, greater than 0.6 mol% and less than 0.7 mol%, greater than 0.7 mol% and less than 0.75 mol%, or greater than 0.75 mol% and less than 0.8 mol%.

[0150] In some embodiments, the proportion of the polymeric lipid (e.g., PEG lipid) in the above-described LNPs to the total lipid present in the LNP is: greater than 0.01% and less than 0.05%, greater than 0.05% and less than 0.07%, greater than 0.07% and less than 0.1%, greater than 0.1% and less than 0.15%, greater than 0.15% and less than 0.2%, greater than 0.2% and less than 0.25%, greater than 0.25% and less than 0.3%, greater than 0.3% and less than 0.35%, greater than 0.35% and less than 0.4%, greater than 0.4% and less than 0.45%, or greater than 0.45% and less than 0.5%.

[0151] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.00011 mol% to 0.00049 mol%, 0.0051% to 0.0099%, 0.011% to 0.0499%, 0.00051 mol% to 0.00099 mol%, 0.0011 mol% to 0.0049 mol%, 0.051 mol% to 0.0999 mol%, 0.101 mol% to 0.149 mol%, 0.151 mol% to 0.199 mol%, 0.205 mol% to 0.249 mol%, 0.255 mol% to 0.299 mol%, 0.305 mol% to 0.349 mol%, 0.355 mol% to 0.399 mol%, 0.405 mol% to 0.449 mol%, 0.455 mol% to 0.499 mol%, 0.501 mol% to 0.549 mol%, 0.551 mol% to 0.599 mol%, 0.601 mol% to 0.649 mol%, 0.651 mol% to 0.699 mol%, 0.701 mol% to 0.749 mol%, or 0.751 mol% to 0.799 mol% of the total lipid present in the LNP.

[0152] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.011% to 0.0499%, 0.051% to 0.0999%, 0.101% to 0.149%, 0.151% to 0.199%, 0.205% to 0.249%, 0.255% to 0.299%, 0.305% to 0.349%, 0.355% to 0.399%, 0.405% to 0.449%, or 0.455% to 0.499% of the total lipid present in the LNP.

[0153] In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.00015 mol% to 0.00045 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.00055 mol% to 0.00095 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.0015 mol% to 0.045 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.055 mol% to 0.095 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.11 mol% to 0.145 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.155 mol% to 0.195 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.21 mol% to 0.245 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.255 mol% to 0.295 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.31 mol% to 0.345 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.355 mol% to 0.395 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.41 mol% to 0.445 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.455 mol% to 0.495 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.51 mol% to 0.545 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.551 mol% to 0.595 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP is present at 0.61 mol% to 0.645 mol% of the total lipid present in the LNP.In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP comprises 0.655 mol% to 0.695 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP comprises 0.71 mol% to 0.745 mol% of the total lipid present in the LNP. In some embodiments, the polymeric lipid (e.g., PEG lipid) in the above LNP comprises 0.755 mol% to 0.795 mol% of the total lipid present in the LNP.

[0154] The term "ionizable lipid" refers to a lipid that becomes positively charged when the pH is lowered below the pKa of the ionizable group of the lipid, but gradually becomes neutral at higher pH values. The positively charged lipid is capable of binding to a negatively charged nucleic acid when the pH is below the pKa. In certain embodiments, the ionizable lipid comprises a zwitterionic lipid.

[0155] In some embodiments, the ionizable lipid of the above LNP comprises the following compound (I), N-oxide thereof, salt thereof, or isomer thereof:

[0156] wherein:

[0157] R1is selected from the group consisting of C5-C 30 alkyl, C5-C 20 alkenyl, -R*YR”, -YR”, and -R”’M’R’;

[0158] R2and R3are independently selected from the group consisting of H, C1-C 14 alkyl, C2-C 14 alkenyl, -R*YR”, -YR”, and -R*OR”, or R2and R3together with the atoms to which they are attached form a heterocyclic or carbocyclic ring;

[0159] R4is selected from the group consisting of hydrogen, C3-C6carbocyclic, -(CH2) n Q, -(CH2) n CHQR, -(CH2) o C(R 10 )2(CH2) n-o Q, -CHQR, -CQ(R)2, and unsubstituted C1-C6alkyl, wherein Q is selected from carbocyclic, heterocyclic, -OR, -O(CH2) nN(R)2, -C(O)OR, -OC(O)R, -CX3, -CX2H, -CXH2, -CN, -N(R)2, -C(O)N(R)2, -N(R)C(O)R, -N(R)S(O)2R, -N(R)C(O)N(R)2, -N(R)C(S)N(R)2, N(R)R8, -N(R)S(O)2R8, -O(CH2) n OR, -N(R)C(=NR9)N(R)2, -N(R)C(=CHR9)N(R)2, -OC(O)N(R)2, -N(R)C(O)OR, -N(OR)C(O)R, -N(OR)S(O)2R, -N(OR)C(O)OR, -N(OR)C(O)N(R)2, -N(OR)C(S)N(R)2, -N(OR)C(=NR9)N(R)2, -N(OR)C(=CHR9)N(R)2, -C(=NR9)N(R)2, -C(=NR9)R, -C(O)N(R)OR, and -C(R)N(R)2C(O)OR, each o is independently selected from 1, 2, 3, and 4, and each n is independently selected from 1, 2, 3, 4, and 5;

[0160] each R5is independently selected from the group consisting of OH, C1-C3alkyl, C2-C3alkenyl, and H;

[0161] each R6is independently selected from the group consisting of OH, C1-C3alkyl, C2-C3alkenyl, and H;

[0162] M and M’ are independently selected from -C(O)O-, -OC(O)-, -OC(O)-M”-C(O)O-, -C(O)N(R’)-, -N(R’)C(O)-, -C(O)-, -C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR’)O-, -S(O)2-, -S-S-, an aryl group, and a heteroaryl group, wherein M” is a bond, C1-C 13 alkylene, or C 2- -C 13 alkylene;

[0163] R7is selected from the group consisting of C 1-3 alkyl, C2-C3alkenyl, and H;

[0164] R8is selected from the group consisting of C 3-6 carbocycle, and heterocycle;

[0165] R9is selected from the group consisting of H, CN, NO2, C1-C6alkyl, -OR, -S(O)2R, -S(O)2N(R)2, C2-C6alkenyl, C3-C6carbocycle, and heterocycle;

[0166] R 10 is selected from the group consisting of H, C1-C3 alkyl, and C2-C3 alkenyl;

[0167] each R is independently selected from the group consisting of C1-C3 alkyl, C2-C3 alkenyl, (CH2) q OR** and H,

[0168] and each q is independently selected from 1, 2, and 3;

[0169] each R’ is independently selected from the group consisting of C1-C 18 alkyl, C2-C 18 alkenyl, -R*YR”, -YR”, H and and R 11 is selected from the group consisting of C1-C 12 alkylene and C2-C 12 alkenylene, R 12 and R 13 are each independently selected from the group consisting of C1-C 12 alkyl and C2-C 12 alkenyl;

[0170] each R” is independently selected from the group consisting of C3-C 15 alkyl and C3-C 15 alkenyl;

[0171] each R”’ is independently selected from the group consisting of C3-C 15 alkylene and C3-C 15 alkenylene;

[0172] each R* is independently selected from the group consisting of absent, C1-C 12 alkylene and C2-C 12 alkenylene;

[0173] each R** is independently selected from the group consisting of absent, C1-C 12 alkyl and C2-C 12 alkenyl;

[0174] each Y is independently a C3-C6 carbocyclic ring;

[0175] each X is independently selected from the group consisting of F, Cl, Br, and I; and

[0176] m is selected from 5, 6, 7, 8, 9, 10, 11, 12, and 13; and wherein when R4 is -(CH2) n Q, -(CH2) nwhen CHQR, -CHQR, or -CQ(R)2, then (i) when n is 1, 2, 3, 4, or 5, Q is other than -N(R)2; or (ii) when n is 1 or 2, Q is other than 5-, 6-, or 7-membered heterocycloalkyl.

[0177] In some embodiments, the ionizable lipids of the above LNP comprise the following compound (II), N-oxide thereof, salt thereof, or isomer thereof:

[0178] wherein:

[0179] R1, R2, R3, R5, R6, M, and R7are as described above,

[0180] R N is H or C1-C3 alkyl;

[0181] X a and X b are each independently O or S;

[0182] R 14 is selected from H, halogen, -OH, R b , -N(R b )2, -CN, -N3, -C(O)OH, -C(O)OR b , -OC(O)R b , -OR b , -SR b , -S(O)R b , -S(O)OR b , -S(O)2OR b , -NO2, -S(O)2N(R b )2, -N(R b )S(O)2R b , -NH(CH2) t1 N(R b )2, -NH(CH2) p1 O(CH2) q1 N(R b )2, -NH(CH2) s1 OR b , -N((CH2) S OR b )2, -N(R b )-carbocyclic ring, -N(R b )-heterocyclic ring, -N(R b )-aryl, -N(R b )-heteroaryl, -N(R b )(CH2) t1 -carbocyclic ring, -N(R b )(CH2)t1 -heteroaryl, carbocycle, heterocycle, aryl, and heteroaryl; b (CH2) t1- (CH2) b (CH2) t1 - heteroaryl, carbocycle, heterocycle, aryl, and heteroaryl;

[0183] each R b is independently selected from the group consisting of C1-C3 alkyl, C2-C3 alkenyl, and H;

[0184] u is 5, 6, 7, 8, 9, 10, 11, 12, or 13;

[0185] w is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;

[0186] r is 0 or 1;

[0187] t 1 is 1, 2, 3, 4, or 5;

[0188] p 1 is 1, 2, 5, 4, or 5;

[0189] q 1 is 1, 2, 5, 4, or 5; and

[0190] s 1 is 1, 2, 3, 4, or 5.

[0191] In some embodiments, the ionizable lipid of the above LNP comprises the following compound (III), an N-oxide thereof, a salt thereof, or an isomer thereof:

[0192] wherein:

[0193] L 1 or L 2 is -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O) x -, -S-S-, -C(=O)S-, -SC(=O)-, -NR a C(=O)-, -C(=O)NR a -, NR a C(=O)NR a -, -OC(=O)NR a -, or -NR a C(=O)O-, and the other of L 1 or L 2 is -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O) x-, -SS-, -C(=O)S-, SC(=O)-, -NR a C(=O)-、-C(=O)NR a -、NR a C(=O)NR a -、-OC(=O)NR a -or-NR a C(=O)O- or bond;

[0194] G 1 and G 2 Each is independently unsubstituted C1-C 12 Alkylene or C2-C 12 alkenyl;

[0195] G 3 For C1-C 24 Alkylene, C2-C 24 C3-C8 cycloalkylene, C3-C8 cycloalkylene;

[0196] R a For H or C1-C 12 hydrocarbon group;

[0197] R 15 and R 16 Each independently is C6-C 24 Alkyl or C6-C 24 alkenyl;

[0198] R 17 For H, OR 18 CN, -C(=O)OR 19 -OC(=O)R 19 Or –NR 18 C(=O)R 19 ;

[0199] R 19 For C1-C 12 hydrocarbon group;

[0200] R 18 It is an H or C1-C6 hydrocarbon group; and

[0201] x is 0, 1, or 2.

[0202] In some embodiments, the ionizable lipids of the above-mentioned LNPs include or are the following compounds (VI), their N-oxides, their salts, or isomers thereof:

[0203] in

[0204] A1, A2, and A3 fall under one of the following categories:

[0205] (1) when the dashed line connecting A1and A2and the dashed line connecting A1and A3are absent, A1is H, C1-C5alkyl or C1-C5heteroalkyl, A2is H, C1-C5alkyl or C1-C5heteroalkyl, A3is C1-C5alkylene or a bond;

[0206] (2) when the dashed line connecting A1and A2is a bond and the dashed line connecting A1and A3is absent, A1is C1-C5alkylene or C1-C5heteroalkylene, A2is C1-C5alkylene or C1-C5heteroalkylene, A3is C1-C5alkylene or a bond, A1and A2together with the nitrogen atom to which they are both attached form an N-containing heterocyclic ring; or

[0207] (3) when the dashed line connecting A1and A3is a bond and the dashed line connecting A1and A2is absent, A1is C1-C5alkylene or C1-C5heteroalkylene, A2is H, C1-C5alkyl or C1-C5heteroalkyl, A3is C1-C5alkylene, A1and A3together with the nitrogen atom to which they are both attached form an N-containing heterocyclic ring;

[0208] (4) when the dashed line connecting A1and A2and the dashed line connecting A1and A3are a bond, A1is C1-C5alkylene or C1-C5heteroalkylene, A2is C1-C5alkylene or C1-C5heteroalkylene, A3is C1-C5alkylene, A1, A2and A3together with the nitrogen atom to which they are all attached form an N-containing spirocyclic, fused or bridged heterocyclic ring;

[0209] A4is C1-C5alkylene or a bond;

[0210] A5, A6, A7and A8are each independently C1-C 18 alkylene, C1-C 18 heteroalkylene or a bond;

[0211] Z1and Z2are each independently -C(=O)O-, -OC(=O)-, -O-, -C(=O)-, -S-, -C(=O)S-, -SC(=O)-, -C(=O)N(R 22 )-, -N(R 22 )C(=O)-, -OC(=O)O-, -OC(=O)S-, -SC(=O)O-, -SC(=O)S-, -N(R 22 )C(=O)O-, -OC(=O)N(R 22 )-, -N(R 22 )C(=O)N(R 22 )-, -SC(=O)N(R 22 )-, -N(R 22)C(=O)S-、-N(C(=O)A9OA 10 )-、-N(C(=O)A9SA 10 )-、-N(C(=O)A 11 ), -N(A9OA 10 )-、-N(A9SA 10 )-、-N(A 11 )-, -OS(=O)O-, -OS(=O)2O-, -OP(=O)O-, -OP(=O)(OH)O-, -OP(=O)(H)O-, -OS(=O)2NH- or -NHS(=O)2O-;

[0212] R 22 Each is independently an H or C1-C8 hydrocarbon group;

[0213] Each R 20 and each R 21 Each independently is C1-C 24 Hydrocarbon group or C1-C containing O or S 24 heterohydrocarbon group;

[0214] Each A9 group is a C1-C8 hydrocarbon group or bond;

[0215] Each A 10 and each A 11 Each is independently a C1-C8 hydrocarbon group;

[0216] In some embodiments, the hydrocarbon group is alkyl, alkenyl, or ynyl;

[0217] In some embodiments, the alkylene group is an alkylene group, an alkenylene group, or an alkynylene group;

[0218] In some embodiments, the heteroalkyl group is a heteroalkyl, heteroalkenyl, or heteroynyl group;

[0219] In some embodiments, the heteroalkyl group is a heteroalkyl group, a heteroalkenyl group, or a heteroyne group.

[0220] In some embodiments, the ionizable lipids of the above-mentioned LNPs are one or more of the following: SM-102, AL-0315, MC3, and

[0221] In some embodiments, the ionizable lipid of the above LNPs does not include SM-102, AL-0315, Dlin-DMA, Dlin-D-DMA, DLin-MC3-DMA (i.e., MC3), DODMA, DSDMA, DLen-DMA, g-DLen-DMA, Dlin-K-DMA, Dlin-KC2-DMA (i.e., Dlin-C2K-DMA, XTC2 or C2K), Dlin-K-C3-DMA, Dlin-K-C4-DMA, g-Dlen-K-C2K-DMA, Dlin-M-C2-DMA (i.e., MC2), Dlin-MP-DMA (i.e., 1-B11), DODAP, or DOTAP.

[0222] In some embodiments, the ionizable lipid in the above LNPs is present at 25 mol% to 75 mol% of the total lipid present in the LNP. For example, 25 mol%, 28 mol%, 30 mol%, 31 mol%, 32 mol%, 33 mol%, 34 mol%, 35 mol%, 36 mol%, 37 mol%, 38 mol%, 39 mol%, 40 mol%, 41 mol%, 42 mol%, 43 mol%, 44 mol%, 45 mol%, 45.5 mol%, 46 mol%, 46.5 mol%, 47 mol%, 47.5 mol%, 48 mol%, 48.5 mol%, 49 mol%, 49.5 mol%, 50 mol%, 50.5 mol%, 51 mol%, 52.5 mol%, 53 mol%, 53.5 mol%, 54 mol%, 54.5 mol%, 55 mol%, 55.5 mol%, 56 mol%, 56.5 mol%, 57 mol%, 57.5 mol%, 58 mol%, 58.5 mol%, 59 mol%, 59.5 mol%, 60 mol%, 60.5 mol%, 61 mol%, 61.5 mol%, 62 mol%, 62.5 mol%, 63 mol%, 63.5 mol%, 64 mol%, 64.5 mol%, 65 mol%, 68 mol%, 70 mol%, or 75 mol%.

[0223] In some embodiments, the ionizable lipid in the above LNP comprises 30-65 mol%, 35-65 mol%, 40-65 mol%, 45-65 mol%, 46-65 mol%, 47-65 mol%, 48-65 mol%, 49-65 mol%, 50-65 mol%, 51-65 mol%, 52-65 mol%, 53-65 mol%, 54-65 mol%, 55-65 mol%, 56-65 mol%, 57-65 mol%, 30-60 mol%, 35-60 mol%, 40-60 mol%, 45-60 mol%, 46-60 mol%, 46.5-60 mol%, 47-60 mol%, 47.5-60 mol%, 48-60 mol%, 48.5-60 mol%, 49-60 mol%, 49.5-60 mol%, 50-60 mol%, 50.5-60 mol%, 51-60 mol%, 51.5-60 mol%, 52.5-60 mol%, 52-60 mol%, 53-60 mol%, 53.5-60 mol%, 54-60 mol%, 54.5-60 mol%, 55-60 mol%, 56-60 mol%, 57-60 mol%, 30-58 mol%, 35-58 mol%, 40-58 mol%, 45-58 mol%, 46-58 mol%, 46.5-58 mol%, 47-58 mol%, 47.5-58 mol%, 48-58 mol%, 48.5-58 mol%, 49-58 mol%, 49.5-58 mol%, 50-58 mol%, 50.5-58 mol%, 51-58 mol%, 51.5-58 mol%, 52-58 mol%, 52.5-58 mol%, 53-58 mol%, 53.5-58 mol%, 54-58 mol%, 54.5-58 mol%, 55-58 mol%, 56-58 mol%, 57-58 mol%, 30-55 mol%, 35-55 mol%, 40-55 mol%, 45-55 mol%, 46-55 mol%, 46.5-55 mol%, 47-55 mol%, 47.5-55 mol%, 48-55 mol%, 48.5-55 mol%, 49-55 mol%, 49.5-55 mol%, 50-55 mol%, 50.5-55 mol%, 51-55 mol%, 51.5-55 mol%, 52-55 mol%, 52.5-55 mol%, 53-55 mol%, 53.5-55 mol%, 54-55 mol%, 54.5-55 mol%, 55-55 mol% of the total lipid present in the LNP.5mol%~58mol%、55mol%~58mol%、56mol%~60mol%、57mol%~58mol%、30mol%~56mol%、35mol%~56mol%、40mol%~56mol%、45mol%~56mol%、46mol%~56mol%、46.5mol%~56mol%、47mol%~56mol%、47.5mol%~56mol%、48mol%~56mol%、48.5mol%~56mol%、49mol%~56mol%、49.5mol%~56mol%、50mol%~56mol%、50.5mol%~56mol%、51mol%~56mol%、51.5mol%~56mol%、52mol%~56mol%、52.5mol%~56mol%、53mol%~56mol%、53.5mol%~56mol%、54mol%~56mol%、54.5mol%~56mol%、55mol%~56mol%、30mol%~55mol%、35mol%~55mol%、40mol%~55mol%、45mol%~55mol%、46mol%~55mol%、46.5mol%~55mol%、47mol%~55mol%、47.5mol%~55mol%、48mol%~55mol%、48.5mol%~55mol%、49mol%~55mol%、49.5mol%~55mol%、50mol%~55mol%、50.5mol%~55mol%、51mol%~55mol%、51.5mol%~55mol%、52mol%~55mol%、52.5mol%~55mol%、53mol%~55mol%、53.5mol%~55mol%、54mol%~55mol%、54.5mol%~55mol%、30mol%~54mol%、35mol%~54mol%、40mol%~54mol%、45mol%~54mol%、46mol%~54mol%、46.5mol%~54mol%、47mol%~54mol%、47.5mol%~54mol%、48mol%~54mol%、48.5mol%~54mol%、49mol%~54mol%、49.5mol%~54mol%、50mol%~54mol%、50.5mol%~54mol%、51mol%~54mol%、51.5mol%~54mol%、52mol%~54mol%、52.5-54, 53.5-54, 30-53, 35-53, 40-53, 45-53, 46-53, 46.5-53, 47-53, 47.5-53, 48-53, 48.5-53, 49-53, 49.5-53, 50-53, 50.5-53, 51-53, 51.5-53, 52-53, 52.5-53, 30-52, 35-52, 40-52, 45-52, 46-52, 47-52, 48-52, 48.5-52, 49-52, 49.5-52, 50-52, 50.5-52, 51-52, 51.5-52, 30-51, 35-51, 40-51, 45-51, 46-51, 47-51, 48-51, 48.5-51, 49-51, 49.5-51, 50-51, or 50.5-51 mol%.

[0224] In some embodiments, the ionizable lipid in the above LNPs comprises 32-59.5 mol% of the total lipid present in the LNP.

[0225] In some embodiments, the ionizable lipid in the above LNPs comprises 35-59.5 mol% of the total lipid present in the LNP.

[0226] In some embodiments, the ionizable lipid in the above LNPs comprises 40-56 mol% of the total lipid present in the LNP.

[0227] In some embodiments, the ionizable lipid in the above LNPs comprises 45-55 mol% of the total lipid present in the LNP.

[0228] In some embodiments, the ionizable lipid in the above LNP comprises 48-55 mol% of the total lipid present in the LNP.

[0229] In some embodiments, the ionizable lipid in the above LNP comprises 49-52 mol% of the total lipid present in the LNP.

[0230] In some embodiments, the ionizable lipid in the above LNP comprises 49.5-51.5 mol% of the total lipid present in the LNP.

[0231] In some embodiments, the helper lipid of the above LNP comprises a phospholipid substance (phospholipid). Phospholipid substances are typically semi-synthetic, and can also be of natural origin or chemically modified. In one alternative specific example, the helper lipid of the above LNP is a phospholipid substance.

[0232] In some embodiments, the phospholipid substance of the above LNP comprises one or more of DSPC (distearoylphosphatidylcholine), DOPE (dioleoylphosphatidylethanolamine), DOPC (dioleoylphosphatidylcholine), DOPS (dioleoylphosphatidylserine), DSPG (1,2-dioctadecanoyl-sn-glycero-3-phospho-(1’-rac-glycerol)), DPPG (dipalmitoylphosphatidylglycerol), DPPC (dipalmitoylphosphatidylcholine), DGTS (1,2-dipalmitoyl-sn-glycero-3-0-4’-(N,N,N-trimethyl)homoserine), and lysophospholipid. In some embodiments, the helper lipid of the above LNP is selected from one or more of DSPC, DOPE, DOPC, and DOPS. In other embodiments, the above LNP does not contain DOPE.

[0233] In some embodiments, the helper lipid of the above LNP is DSPC and / or DOPE.

[0234] In some embodiments, the helper lipid of the above LNP is DSPC.

[0235] In some embodiments, the helper lipid (e.g., DSPC) in the above LNP is present at 0-45 mol% of the total lipid present in the LNP. For example, 0.5 mol%, 1 mol%, 3 mol%, 5 mol%, 7.5 mol%, 8 mol%, 8.5 mol%, 9 mol%, 9.5 mol%, 10 mol%, 10.5 mol%, 11 mol%, 11.5 mol%, 12 mol%, 12.5 mol%, 13 mol%, 13.5 mol%, 14 mol%, 14.5 mol%, 15 mol%, 15.5 mol%, 16 mol%, 16.5 mol%, 17 mol%, 17.5 mol%, 18 mol%, 18.5 mol%, 19 mol%, 19.5 mol%, 20 mol%, 20.5 mol%, 21 mol%, 21.5 mol%, 22 mol%, 22.5 mol%, 23 mol%, 23.5 mol%, 24 mol%, 24.5 mol%, 25 mol%, 25.5 mol%, 26 mol%, 26.5 mol%, 27 mol%, 27.5 mol%, 28 mol%, 28.5 mol%, 29 mol%, 29.5 mol%, 30 mol%, 34 mol%, 35 mol%, 36 mol%, 38 mol%, 40 mol%, 42 mol%, 44 mol%, or 45 mol%.

[0236] In some embodiments, the helper lipid (e.g., DSPC) in the above LNP is present at 0-30 mol% of the total lipid present in the LNP.

[0237] In some embodiments, the helper lipid (e.g., DSPC) in the above LNP is present at a ratio greater than 0 and less than 30 mol% of the total lipid present in the LNP.

[0238] In some embodiments, the helper lipid (e.g., DSPC) in the above LNPs is present at 0.5 mol% to 29.5 mol%, 0.5 mol% to 27.5 mol%, 1.5 mol% to 27.5 mol%, 2.5 mol% to 27.5 mol%, 4.5 mol% to 27.5 mol%, 5.5 mol% to 27.5 mol%, 6.5 mol% to 27.5 mol%, 7.5 mol% to 27.5 mol%, 8 mol% to 27.5 mol%, 8.5 mol% to 27.5 mol%, 9 mol% to 27.5 mol%, 9.5 mol% to 27.5 mol%, 10 mol% to 27.5 mol%, 10.5 mol% to 27.5 mol%, 11 mol% to 27.5 mol%, 11.5 mol% to 27.5 mol%, 12 mol% to 27.5 mol%, 12.5 mol% to 27.5 mol%, 13 mol% to 27.5 mol%, 13.5 mol% to 27.5 mol%, 14 mol% to 27.5 mol%, 14.5 mol% to 27.5 mol%, 15 mol% to 27.5 mol%, 15.5 mol% to 27.5 mol%, 16 mol% to 27.5 mol%, 16.5 mol% to 27.5 mol%, 17 mol% to 27.5 mol%, 17.5 mol% to 27.5 mol%, 18 mol% to 27.5 mol%, 18.5 mol% to 27.5 mol%, 19 mol% to 27.5 mol%, 20 mol% to 27.5 mol%, 21 mol% to 27.5 mol%, 0.5 mol% to 25 mol%, 2.5 mol% to 25 mol%, 4.5 mol% to 25 mol%, 5.5 mol% to 25 mol%, 6.5 mol% to 25 mol%, 7.5 mol% to 25 mol%, 8 mol% to 25 mol%, 8.5 mol% to 25 mol%, 9 mol% to 25 mol%, 9.5 mol% to 25 mol%, 10 mol% to 25 mol%, 10.5 mol% to 25 mol%, 11 mol% to 25 mol%, 11.5 mol% to 25 mol%, 12 mol% to 25 mol%, 12.5 mol% to 25 mol%, 13 mol% to 25 mol%, 13.5 mol% to 25 mol%, 14 mol% to 25 mol%, 14.5 mol% to 25 mol%, 15 mol% to 25 mol%, 15.5 mol% to 25 mol%, 16 mol% to 25 mol%, 16.5 mol% to 25 mol%, 17 mol% to 25 mol%, 17.5 mol% to 25 mol%, 18 mol% to 25 mol%, 18.5 mol% to 25 mol%, 19 mol% to 25 mol%, 20 mol% to 25 mol%, 21 mol% to 25 mol%, 22 mol% to 25 mol%, 23 mol% to 25 mol%, 24 mol% to 25 mol%, 24.5 mol% to 25 mol%, 25 mol% to 30 mol%, 25 mol% to 35 mol%, 25 mol% to 40 mol%, 25 mol% to 45 mol%, 25 mol% to 50 mol%, 25 mol% to 55 mol%, 25 mol% to 60 mol%, 25 mol% to 65 mol%, 25 mol% to 70 mol%, 25 mol% to 75 mol%, 25 mol% to 80 mol%, 25 mol% to 85 mol%, 25 mol% to 90 mol%, 25 mol% to 95 mol%, 25 mol% to 100 mol%, 25 mol% to 105 mol%, 25 mol% to 110 mol%, 25 mol% to 115 mol%, 25 mol% to 120 mol%, 25 mol% to 125 mol%, 25 mol% to 130 mol%, 25 mol% to 135 mol%, 25 mol% to 140 mol%, 25 mol% to 145 mol%, 25 mol% to 150 mol%, 25 mol% to 155 mol%, 25 mol% to 160 mol%, 25 mol% to 165 mol%, 25 mol% to 170 mol%, 25 mol% to 175 mol%, 25 mol% to 180 mol%, 25 mol% to 185 mol%, 25 mol% to 190 mol%, 25 mol% to 195 mol%, 25 mol% to 200 mol%, 25 mol% to 205 mol%, 25 mol% to 210 mol%, 25 mol% to 215 mol%, 25 mol% to 220 mol%, 25 mol% to 225 mol%, 25 mol% to 230 mol%, 25 mol% to 235 mol%, 25 mol% to 240 mol%, 25 mol% to 245 mol%, 25 mol% to 250 mol%, 25 mol% to 255 mol%, 25 mol% to 260 mol%, 25 mol% to 265 mol%, 25 mol% to 270 mol%, 25 mol% to 275 mol%, 25 mol% to 280 mol%, 25 mol% to 285 mol%, 25 mol% to 290 mol%, 25 mol% to 295 mol%, or 25 mol% to 300 mol% of the total lipid present in the LNP.5 mol% to 25 mol%, 19 mol% to 25 mol%, 19.5 mol% to 25 mol%, 20 mol% to 25 mol%, 21 mol% to 25 mol%, 0.5 mol% to 23 mol%, 2.5 mol% to 23 mol%, 4.5 mol% to 23 mol%, 5.5 mol% to 23 mol%, 6.5 mol% to 23 mol%, 7.5 mol% to 23 mol%, 8 mol% to 23 mol%, 8.5 mol% to 23 mol%, 9 mol% to 23 mol%, 9.5 mol% to 23 mol%, 10 mol% to 23 mol%, 10.5 mol% to 23 mol%, 11 mol% to 23 mol%, 11.5 mol% to 23 mol%, 12 mol% to 23 mol%, 12.5 mol% to 23 mol%, 13 mol% to 23 mol%, 13.5 mol% to 23 mol%, 14 mol% to 23 mol%, 14.5 mol% to 23 mol%, 15 mol% to 23 mol%, 15.5 mol% to 23 mol%, 16 mol% to 23 mol%, 16.5 mol% to 23 mol%, 17 mol% to 23 mol%, 17.5 mol% to 23 mol%, 18 mol% to 23 mol%, 18.5 mol% to 23 mol%, 19 mol% to 23 mol%, 20 mol% to 23 mol%, 21 mol% to 23 mol%, 0.5 mol% to 20 mol%, 2.5 mol% to 20 mol%, 4.5 mol% to 20 mol%, 5.5 mol% to 20 mol%, 6.5 mol% to 20 mol%, 7.5 mol% to 20 mol%, 8 mol% to 20 mol%, 8.5 mol% to 20 mol%, 9 mol% to 20 mol%, 9.5 mol% to 20 mol%, 10 mol% to 20 mol%, 10.5 mol% to 20 mol%, 11 mol% to 20 mol%, 11.5 mol% to 20 mol%, 12 mol% to 20 mol%, 12.5 mol% to 20 mol%, 13 mol% to 20 mol%, 13.5 mol% to 20 mol%, 14 mol% to 20 mol%, 14.5 mol% to 20 mol%, 15 mol% to 20 mol%, 15.5 mol% to 20 mol%, 16 mol% to 20 mol%, 16.5 mol% to 20 mol%, 17 mol% to 20 mol%, 17.5 mol% to 20 mol%, 18 mol% to 20 mol%, 18.5 mol% to 20 mol%, 19 mol% to 20 mol%, 0.5 mol% to 18 mol%, 2.5 mol% to 18 mol%, 4.5 mol% to 18 mol%, 5.5 mol% to 18 mol%, 6.5 mol% to 18 mol%, 7.5 mol% to 18 mol%, 8 mol% to 18 mol%, 8.5 mol% to 18 mol%, 9 mol% to 18 mol%, 9.5 mol% to 18 mol%, 10 mol% to 18 mol%, 10.5 mol% to 18 mol%, 11 mol% to 18 mol%, 11.5 mol% to 18 mol%, 12 mol% to 18 mol%, 12.5 mol% to 18 mol%, 13 mol% to 18 mol%, 13.5 mol% to 18 mol%, 14 mol% to 18 mol%, 14.5 mol% to 18 mol%, 15 mol% to 18 mol%, 15.5 mol% to 18 mol%, 16 mol% to 18 mol%, 16.5 mol% to 18 mol%, 17 mol% to 18 mol%, 17.5 mol% to 18 mol%, 0.5 mol% to 17 mol%, 2.5 mol% to 17 mol%, 4.5 mol% to 17 mol%, 5.5 mol% to 17 mol%, 6.5 mol% to 17 mol%, 7.5 mol% to 17 mol%, 8 mol% to 17 mol%, 8.5 mol% to 17 mol%, 9 mol% to 17 mol%, 9.5 mol% to 17 mol%, 10 mol% to 17 mol%, 10.5 mol% to 17 mol%, 11 mol% to 17 mol%, 11.5 mol% to 17 mol%, 12 mol% to 17 mol%, 12.5 mol% to 17 mol%, 13 mol% to 17 mol%, 13.5 mol% to 17 mol%, 14 mol% to 17 mol%, 14.5 mol% to 17 mol%, 15 mol% to 17 mol%, 15.5 mol% to 17 mol%, 16 mol% to 17 mol%, 16.5 mol% to 17 mol%, 0.5 mol% to 16 mol%, 2.5 mol% to 16 mol%, 4.5 mol% to 16 mol%, 5.5 mol% to 16 mol%, 6.5 mol% to 16 mol%, 7.5 mol% to 16 mol%, 8 mol% to 16 mol%, 8.5 mol% to 16 mol%, 9 mol% to 16 mol%, 9.5 mol% to 16 mol%, 10 mol% to 16 mol%, 10.5 mol% to 16 mol%, 11 mol% to 16 mol%, 11.5 mol% to 16 mol%, 12 mol% to 16 mol%, 12.5 mol% to 16 mol%, 13 mol% to 16 mol%, 13.5 mol% to 16 mol%, 14 mol% to 16 mol%, 14.5 mol% to 16 mol%, 15 mol% to 16 mol%, 15.5 mol% to 16 mol%, 16 mol% to 16 mol%,5 mol% to 16 mol%, 15 mol% to 16 mol%, or 15.5 mol% to 16 mol%.

[0239] In some embodiments, the helper lipid (e.g., DSPC) in the above LNP comprises 7.5 mol% to 25 mol% of the total lipid present in the LNP.

[0240] In some embodiments, the helper lipid (e.g., DSPC) in the above LNP comprises 8 mol% to 20 mol% of the total lipid present in the LNP.

[0241] In some embodiments, the helper lipid (e.g., DSPC) in the above LNP comprises 10 mol% to 18 mol% of the total lipid present in the LNP.

[0242] In some embodiments, the helper lipid (e.g., DSPC) in the above LNP comprises 12 mol% to 16 mol% of the total lipid present in the LNP.

[0243] In some embodiments, the structural lipid of the above LNP comprises a sterol. In an alternative specific example, the structural lipid of the above LNP is a sterol. In some embodiments, the sterol of the above LNP comprises one or more of the following: phytosterol, 20a-hydroxycholesterol, cholesterol, cholesterol ester, sterol hormone, sterol vitamin, bile acid, cholestanol, ergosterol, beta-sitosterol, and oxidized cholesterol derivative. In some embodiments, the structural lipid of the above LNP comprises at least one of the following: cholesterol, cholesterol ester, sterol hormone, sterol vitamin, and bile acid.

[0244] In some embodiments, the structural lipid of the above LNP is not a phytosterol or derivative thereof.

[0245] In some embodiments, the structural lipid of the above LNP is cholesterol. In an alternative specific example, the structural lipid of the above LNP is high purity cholesterol, particularly injection grade high purity cholesterol, such as CHO-HP (produced by AVT).

[0246] In some embodiments, the structural lipid of the above LNP is 20a-hydroxycholesterol.

[0247] In some embodiments, the structural lipid (e.g., cholesterol) in the above-described LNPs is present at 0 mol% to 60 mol% of the total lipid present in the LNP. For example, 0 mol%, 1 mol%, 5 mol%, 8 mol%, 10 mol%, 12 mol%, 14 mol%, 15 mol%, 16 mol%, 18 mol%, 20 mol%, 22 mol%, 24 mol%, 25 mol%, 27 mol%, 27.5 mol%, 28 mol%, 28.5 mol%, 29 mol%, 29.5 mol%, 30 mol%, 30.5 mol%, 31 mol%, 31.5 mol%, 32 mol%, 32.5 mol%, 33 mol%, 33.5 mol%, 34 mol%, 34.5 mol%, 35 mol%, 35.5 mol%, 36 mol%, 36.5 mol%, 37 mol%, 37.5 mol%, 38 mol%, 38.5 mol%, 39 mol%, 39.5 mol%, 40 mol%, 40.5 mol%, 41 mol%, 41.5 mol%, 42 mol%, 42.5 mol%, 43 mol%, 44 mol%, 45 mol%, 46 mol%, 47 mol%, 48 mol%, 49 mol%, 50 mol%, 51 mol%, 52 mol%, 53 mol%, 54 mol%, 55 mol%, 56 mol%, 57 mol%, 58 mol%, 59 mol%, or 60 mol%.

[0248] In some embodiments, the structural lipid (e.g., cholesterol) in the above-described LNPs is present at 10-60 mol%, 11-60 mol%, 13-60 mol%, 15-60 mol%, 17-60 mol%, 19-60 mol%, 21-60 mol%, 23-60 mol%, 25-60 mol%, 26-60 mol%, 26.5-60 mol%, 27-60 mol%, 27.5-60 mol%, 28-60 mol%, 28.5-60 mol%, 29-60 mol%, 29.5-60 mol%, 30-60 mol%, 30.5-60 mol%, 31-60 mol%, 31.5-60 mol%, 32-60 mol%, 32.5-60 mol%, 33-60 mol%, 33.5-60 mol%, 34-60 mol%, 34.5-60 mol%, 35-60 mol%, 35.5-60 mol%, 36-60 mol%, 36.5-60 mol%, 37-60 mol%, 37.5-60 mol%, 38-60 mol%, 38.5-60 mol%, 39-60 mol%, 39.5-60 mol%, 40-60 mol%, 40.5-60 mol%, 41-60 mol%, 41.5-60 mol%, 42-60 mol%, 42.5-60 mol%, 43-60 mol%, 43.5-60 mol%, 44-60 mol%, 44.5-60 mol%, 45-60 mol%, 45.5-60 mol%, 46-60 mol%, 46.5-60 mol%, 47-60 mol%, 47.5-60 mol%, 48-60 mol%, 48.5-60 mol%, 49-60 mol%, 49.5-60 mol%, 50-60 mol%, 50.5-60 mol%, 51-60 mol%, 51.5-60 mol%, 52-60 mol%, 52.5-60 mol%, 53-60 mol%, 53.5-60 mol%, 54-60 mol%, 54.5-60 mol%, 55-60 mol%, 55.5-60 mol%, 56-60 mol%, 56.5-60 mol%, 57-60 mol%, 57.5-60 mol%, 58-60 mol%, 58.5-60 mol%, 59-60 mol%, or 59.5-60 mol% of the total lipid present in the LNP.5 to 60 mol%, 45 to 60 mol%, 46 to 60 mol%, 47 to 60 mol%, 48 to 60 mol%, 49 to 60 mol%, 50 to 60 mol%, 51 to 60 mol%, 52 to 60 mol%, 53 to 60 mol%, 54 to 60 mol%, 55 to 60 mol%, 56 to 60 mol%, 57 to 60 mol%, 58 to 60 mol%, 10 to 55 mol%, 11 to 55 mol%, 13 to 55 mol%, 15 to 55 mol%, 17 to 55 mol%, 19 to 55 mol%, 21 to 55 mol%, 23 to 55 mol%, 25 to 55 mol%, 26 to 55 mol%, 26.5 to 55 mol%, 27 to 55 mol%, 27.5 to 55 mol%, 28 to 55 mol%, 28.5 to 55 mol%, 29 to 55 mol%, 29.5 to 55 mol%, 30 to 55 mol%, 30.5 to 55 mol%, 31 to 55 mol%, 31.5 to 55 mol%, 32 to 55 mol%, 32.5 to 55 mol%, 33 to 55 mol%, 33.5 to 55 mol%, 34 to 55 mol%, 34.5 to 55 mol%, 35 to 55 mol%, 35.5 to 55 mol%, 36 to 55 mol%, 36.5 to 55 mol%, 37 to 55 mol%, 37.5 to 55 mol%, 38 to 55 mol%, 38.5 to 55 mol%, 39 to 55 mol%, 39.5 to 55 mol%, 40 to 55 mol%, 40.5 to 55 mol%, 41 to 55 mol%, 41.5 to 55 mol%, 42 to 55 mol%, 42.5 to 55 mol%, 43 to 55 mol%, 43.5 to 55 mol%, 44 to 55 mol%, 44.5 to 55 mol%, 45 to 55 mol%, 45.5 to 55 mol%, 46 to 55 mol%, 46.5 to 55 mol%, 47 to 55 mol%, 47.5 to 55 mol%, 48 to 55 mol%, 48.5 to 55 mol%, 49 to 55 mol%, 49.5 to 55 mol%, 50 to 55 mol%, 50.5 to 55 mol%, 51 to 55 mol%, 51.5 to 55 mol%, 52 to 55 mol%, 52.5 to 55 mol%, 53 to 55 mol%, 53.5 to 55 mol%, 54 to 55 mol%, 54.5 to 55 mol%, 55 mol%.5 to 55 mol%, 45 to 55 mol%, 46 to 55 mol%, 47 to 55 mol%, 48 to 55 mol%, 49 to 55 mol%, 50 to 55 mol%, 51 to 55 mol%, 52 to 55 mol%, 53 to 55 mol%, 54 to 55 mol%, 10 to 50 mol%, 11 to 50 mol%, 13 to 50 mol%, 15 to 50 mol%, 17 to 50 mol%, 19 to 50 mol%, 21 to 50 mol%, 23 to 50 mol%, 25 to 50 mol%, 26 to 50 mol%, 26.5 to 50 mol%, 27 to 50 mol%, 27.5 to 50 mol%, 28 to 50 mol%, 28.5 to 50 mol%, 29 to 50 mol%, 29.5 to 50 mol%, 30 to 50 mol%, 30.5 to 50 mol%, 31 to 50 mol%, 31.5 to 50 mol%, 32 to 50 mol%, 32.5 to 50 mol%, 33 to 50 mol%, 33.5 to 50 mol%, 34 to 50 mol%, 34.5 to 50 mol%, 35 to 50 mol%, 35.5 to 50 mol%, 36 to 50 mol%, 36.5 to 50 mol%, 37 to 50 mol%, 37.5 to 50 mol%, 38 to 50 mol%, 38.5 to 50 mol%, 39 to 50 mol%, 39.5 to 50 mol%, 40 to 50 mol%, 40.5 to 50 mol%, 41 to 50 mol%, 41.5 to 50 mol%, 42 to 50 mol%, 42.5 to 50 mol%, 43 to 50 mol%, 43.5 to 50 mol%, 44 to 50 mol%, 44.5 to 50 mol%, 45 to 50 mol%, 45.5 to 50 mol%, 46 to 50 mol%, 46.5 to 50 mol%, 47 to 50 mol%, 47.5 to 50 mol%, 48 to 50 mol%, 48.5 to 50 mol%, 49 to 50 mol%, 49.5 to 50 mol%, 50 to 50 mol%, 50.5 to 50 mol%, 51 to 50 mol%, 51.5 to 50 mol%, 52 to 50 mol%, 52.5 to 50 mol%, 53 to 50 mol%, 53.5 to 50 mol%, 54 to 50 mol%, 54.5 to 50 mol%, 55 to 50 mol%, 55.5 to 50 mol%, 56 to 50 mol%, 56.5 to 50 mol%, 57 to 50 mol%, 57.5 to 50 mol%, 58 to 50 mol%, 58.5 to 50 mol%, 59 to 50 mol%, 59.5 to 50 mol%, 60 to 50 mol%, 60.5 to 50 mol%, 61 to 50 mol%, 61.5 to 50 mol%, 62 to 50 mol%, 62.5 to 50 mol%, 63 to 50 mol%, 63.5 to 50 mol%, 64 to 50 mol%, 64.5 to 50 mol%, 65 to 50 mol%, 65.5 to 50 mol%, 66 to 50 mol%, 66.5 to 50 mol%, 67 to 50 mol%, 67.5 to 50 mol%, 68 to 50 mol%, 68.5 to 50 mol%, 69 to 50 mol%, 69.5 to 50 mol%, 70 to 50 mol%, 70.5 to 50 mol%, 71 to 50 mol%, 71.5 to 50 mol%, 72 to 50 mol%, 72.5 to 50 mol%, 73 to 50 mol%, 73.5 to 50 mol%, 74 to 50 mol%, 74.5 to 50 mol%, 75 to 50 mol%, 75.5 to 50 mol%, 76 to 50 mol%, 76.5 to 50 mol%, 77 to 50 mol%, 77.5 to 50 mol%, 78 to 50 mol%, 78.5 to 50 mol%, 79 to5mol%~50mol%、45mol%~50mol%、46mol%~50mol%、47mol%~50mol%、48mol%~50mol%、49mol%~50mol%、10mol%~45mol%、11mol%~45mol%、13mol%~45mol%、15mol%~45mol%、17mol%~45mol%、19mol%~45mol%、21mol%~45mol%、23mol%~45mol%、25mol%~45mol%、26mol%~45mol%、26.5mol%~45mol%、27mol%~45mol%、27.5mol%~45mol%、28mol%~45mol%、28.5mol%~45mol%、29mol%~45mol%、29.5mol%~45mol%、30mol%~45mol%、30.5mol%~45mol%、31mol%~45mol%、31.5mol%~45mol%、32mol%~45mol%、32.5mol%~45mol%、33mol%~45mol%、33.5mol%~45mol%、34mol%~45mol%、34.5mol%~45mol%、35mol%~45mol%、35.5mol%~45mol%、36mol%~45mol%、36.5mol%~45mol%、37mol%~45mol%、37.5mol%~45mol%、38mol%~45mol%、38.5mol%~45mol%、39mol%~45mol%、39.5mol%~45mol%、40mol%~45mol%、40.5mol%~45mol%、41mol%~45mol%、41.5mol%~45mol%、42mol%~45mol%、42.5mol%~45mol%、43mol%~45mol%、43.5mol%~45mol%、44mol%~45mol%、44.5mol%~45mol%、10mol%~42mol%、11mol%~42mol%、13mol%~42mol%、15mol%~42mol%、17mol%~42mol%、19mol%~42mol%、21mol%~42mol%、23mol%~42mol%、25mol%~42mol%、26mol%~42mol%、26.5mol%~42mol%、27mol%~42mol%、27.5mol%~42mol%、28mol%~42mol%、28.5-42, 29-42, 29.5-42, 30-42, 30.5-42, 31-42, 31.5-42, 32-42, 32.5-42, 33-42, 33.5-42, 34-42, 34.5-42, 35-42, 35.5-42, 36-42, 36.5-42, 37-42, 37.5-42, 38-42, 38.5-42, 39-42, 39.5-42, 40-42, 40.5-42, 41-42, 41.5-42, 10-40, 11-40, 13-40, 15-40, 17-40, 19-40, 21-40, 23-40, 25-40, 26-40, 26.5-40, 27-40, 27.5-40, 28-40, 28.5-40, 29-40, 29.5-40, 30-40, 30.5-40, 31-40, 31.5-40, 32-40, 32.5-40, 33-40, 33.5-40, 34-40, 34.5-40, 35-40, 35.5-40, 36-40, 36.5-40, 37-40, 37.5-40, 38-40, 38.5-40, 39-40, 39.5-40, 40-40, 10-41, 11-41, 13-41, 15-41, 17-41, 19-41, 21-41, 23-41, 25-41, 26-41, 26.5-41, 27-41, 27.5-41, 28-41, 28.5-41, 29-41, 29.5-41, 30-41, 30.5-41, 31-41, 31.5-41, 32-41, 32.5-41, 33-41, 33.5-41, 34-41, 34.5-41, 35-41, 35.5-41, 36-41, 36.5-41, 37-41, 37.5-41, 38-41, 38.5-41, 39-41, 39.5-41, 40-41, 40.5-41, 41-41, 10-42, 11-42, 13-42, 15-42, 17-42, 19-42, 21-42, 23-42, 25-42, 26-42, 26.5-42, 27-42, 27.5-42, 28-42, 28.5-42, 29-42, 29.5-42, 30-42, 30.5-42, 31-42, 31.5-42, 32-42, 32.5-42, 33-42, 33.5-42, 34-42, 34.5-42, 35-42, 35.5-42, 36-42, 36.5-42, 37-42, 37.5-42, 38-42, 38.5-42, 39-42, 39.5-42, 40-42, 40.5-42, 41-42, 41.5-42, 10-30, 11-30, 13-30, 15-30, 17-30, 19-30, 21-30, 23-30, 25-30, 26-30, 26.5-30, 27-30, 27.5-30, 28-30, 28.5-30, 29-30, 29.5-30, 30-30, 10-29, 11-29, 13-29, 15-29, 17-29, 19-29, 21-29, 23-29, 25-29, 26-29, 26.5-29, 27-29, 27.5-29, 28-29, 28.5-29, 29-29, 10-28, 11-28, 13-28, 15-28, 17-28, 19-28, 21-28, 23-28, 25-28, 26-28, 26.5-28, 27-28, 27.5-28, 28-28, 10-27, 11-27, 13-27, 15-27, 17-27, 19-27, 21-27, 23-27, 25-27, 26-27, 26.5-27, 27-27, 10-26, 11-26, 13-26, 15-26, 17-26, 19-26, 21-26, 23-26, 25-26, 26-26, 10-25, 11-25, 13-25, 15-25, 17-25, 19-25, 21-25, 23-25, 25-25, 10-24, 11-24, 13-24, 15-24, 17-24, 19-24, 21-24, 23-24, 25-24, 10-23, 11-23, 13-23, 15-23, 17-23, 19-23, 21-23, 23-23, 10-22, 11-22, 13-22, 15-22, 17-22, 19-22, 21-22, 23-22, 10-21, 11-21, 13-21, 15-21, 17-21, 19-21, 21-21, 10-20, 11-20, 13-20, 15-20, 17-20, 19-20, 10-19, 11-19, 13-19, 15-19, 17-19, 19-19, 10-18, 11-18, 13-18, 15-18, 17-18, 18-18, 10-17, 11-17, 13-17, 15-17, 17-17, 10-16, 11-16, 13-16, 15-16, 16-16, 10-15, 11-15, 13-15, 15-15, 10-14, 11-14, 13-14, 14-14, 10-13, 11-13, 13-13, 10-12, 11-12, 12-12, 10-11, 11-11, 10-10, 10-10.5 mol% to 40 mol%, 10 mol% to 38 mol%, 11 mol% to 38 mol%, 13 mol% to 38 mol%, 15 mol% to 38 mol%, 17 mol% to 38 mol%, 19 mol% to 38 mol%, 21 mol% to 38 mol%, 23 mol% to 38 mol%, 25 mol% to 38 mol%, 26 mol% to 38 mol%, 26.5 mol% to 38 mol%, 27 mol% to 38 mol%, 27.5 mol% to 38 mol%, 28 mol% to 38 mol%, 28.5 mol% to 38 mol%, 29 mol% to 38 mol%, 29.5 mol% to 38 mol%, 30 mol% to 38 mol%, 30.5 mol% to 38 mol%, 31 mol% to 38 mol%, 31.5 mol% to 38 mol%, 32 mol% to 38 mol%, 32.5 mol% to 38 mol%, 33 mol% to 38 mol%, 33.5 mol% to 38 mol%, 34 mol% to 38 mol%, 34.5 mol% to 38 mol%, 35 mol% to 38 mol%, 35.5 mol% to 38 mol%, 36 mol% to 38 mol%, 36.5 mol% to 38 mol%, 37 mol% to 38 mol%, 37.5 mol% to 38 mol%, 10 mol% to 35 mol%, 11 mol% to 35 mol%, 13 mol% to 35 mol%, 15 mol% to 35 mol%, 17 mol% to 35 mol%, 19 mol% to 35 mol%, 21 mol% to 35 mol%, 23 mol% to 35 mol%, 25 mol% to 35 mol%, 26 mol% to 35 mol%, 26.5 mol% to 35 mol%, 27 mol% to 35 mol%, 27.5 mol% to 35 mol%, 28 mol% to 35 mol%, 28.5 mol% to 35 mol%, 29 mol% to 35 mol%, 29.5 mol% to 35 mol%, 30 mol% to 35 mol%, 30.5 mol% to 35 mol%, 31 mol% to 35 mol%, 31.5 mol% to 35 mol%, 32 mol% to 35 mol%, 32.5 mol% to 35 mol%, 33 mol% to 35 mol%, 33.5 mol% to 35 mol%, 34 mol% to 35 mol%, 34.5 mol% to 35 mol%, 35 mol% to 35 mol%, 35.5 mol% to 35 mol%, 36 mol% to 35 mol%, 36.5 mol% to 35 mol%, 37 mol% to 35 mol%, 37.5 mol% to 35 mol%, 10 mol% to 33 mol%, 11 mol% to 33 mol%, 13 mol% to 33 mol%, 15 mol% to 33 mol%, 17 mol% to 33 mol%, 19 mol% to 33 mol%, 21 mol% to 33 mol%, 23 mol% to 33 mol%, 25 mol% to 33 mol%, 26 mol% to 33 mol%, 26.5 mol% to 33 mol%, 27 mol% to 33 mol%, 27.5 mol% to 33 mol%, 28 mol% to 33 mol%, 28.5 mol% to 33 mol%, 29 mol% to 33 mol%, 29.5 mol% to 33 mol%, 30 mol% to 33 mol%, 30.5 mol% to 33 mol%, 31 mol% to 33 mol%, 31.5 mol% to 33 mol%, 32 mol% to 33 mol%, 32.5 mol% to 33 mol%, 33 mol% to 33 mol%, 33.5 mol% to 33 mol%, 34 mol% to 33 mol%, 34.5 mol% to 33 mol%, 35 mol% to 33 mol%, 35.5 mol% to 33 mol%, 36 mol% to 33 mol%, 36.5 mol% to 33 mol%, 37 mol% to 33 mol%, 37.5 mol% to 33 mol%, 10 mol% to 31 mol%, 11 mol% to 31 mol%, 13 mol% to 31 mol%, 15 mol% to 31 mol%, 17 mol% to 31 mol%, 19 mol% to 31 mol%, 21 mol% to 31 mol%, 23 mol% to 31 mol%, 25 mol% to 31 mol%, 26 mol% to 31 mol%, 26.5 mol% to 31 mol%, 27 mol% to 31 mol%, 27.5 mol% to 31 mol%, 28 mol% to 31 mol%, 28.5 mol% to 31 mol%, 29 mol% to 31 mol%, 29.5 mol% to 31 mol%, 30 mol% to 31 mol%, 30.5 mol% to 31 mol%, 31 mol% to 31 mol%, 31.5 mol% to 31 mol%, 32 mol% to 31 mol%, 32.5 mol% to 31 mol%, 33 mol% to 31 mol%, 33.5 mol% to 31 mol%, 34 mol% to 31 mol%, 34.5 mol% to 31 mol%, 35 mol% to 31 mol%, 35.5 mol% to 31 mol%, 36 mol% to 31 mol%, 36.5 mol% to 31 mol%, 37 mol% to 31 mol%, 37.5 mol% to 31 mol%, 10 mol% to 29 mol%, 11 mol% to 29 mol%, 13 mol% to 29 mol%, 15 mol% to 29 mol%, 17 mol% to 29 mol%, 19 mol% to 29 mol%, 21 mol% to 29 mol%, 23 mol% to 29 mol%, 25 mol% to 29 mol%, 26 mol% to 29 mol%, 26.5 mol% to 29 mol%, 27 mol% to 29 mol%, 27.5 mol% to 29 mol%, 28 mol% to 29 mol%, 28.5 mol% to 29 mol%, 29 mol% to 29 mol%, 29.5 mol% to 29 mol%, 30 mol% to 29 mol%, 30.5 mol% to 29 mol%, 31 mol% to 29 mol%, 31.5 mol% to 29 mol%, 32 mol% to 29 mol%, 32.5 mol% to 29 mol%, 33 mol% to 29 mol%, 33.5 mol% to 29 mol%, 34 mol% to 29 mol%, 34.5 mol% to 29 mol%, 35 mol% to 29 mol%, 35.5 mol% to 29 mol%, 36 mol% to 29 mol%, 36.5 mol% to 29 mol%, 37 mol% to 29 mol%, 37.5 mol% to 29 mol%, 10 mol% to 27 mol%, 11 mol% to 27 mol%, 13 mol% to 27 mol%, 15 mol% to 27 mol%, 17 mol% to 27 mol%, 19 mol% to 27 mol%, 21 mol% to 27 mol%, 23 mol% to 27 mol%, 25 mol% to 27 mol%, 26 mol% to 27 mol%, 26.5 mol% to 27 mol%, 27 mol% to 27 mol%, 27.5 mol% to 27 mol%, 28 mol% to 27 mol%, 28.5 mol% to 27 mol%, 29 mol% to 27 mol%, 29.5 mol% to 27 mol%, 30 mol% to 27 mol%, 30.5 mol% to 27 mol%, 31 mol% to 27 mol%, 31.5 mol% to 27 mol%, 32 mol% to 27 mol%, 32.5 mol% to 27 mol%, 33 mol% to 27 mol%, 33.5 mol% to 27 mol%, 34 mol% to 27 mol%, 34.5 mol% to 27 mol%, 35 mol% to 27 mol%, 35.5 mol% to 27 mol%, 36 mol% to 27 mol%, 36.5 mol% to 27 mol%, 37 mol% to 27 mol%, 37.5 mol% to 27 mol%, 10 mol% to 25 mol%, 11 mol% to 25 mol%, 13 mol% to 25 mol%, 15 mol% to 25 mol%, 17 mol% to 25 mol%, 19 mol% to 25 mol%, 21 mol% to 25 mol%, 23 mol% to 25 mol%, 25 mol% to 25 mol%, 25.5 mol% to 25 mol%, 26 mol% to 25 mol%, 26.5 mol% to 25 mol%, 275 mol% to 35 mol%, 10 mol% to 30 mol%, 11 mol% to 30 mol%, 13 mol% to 30 mol%, 15 mol% to 30 mol%, 17 mol% to 30 mol%, 19 mol% to 30 mol%, 21 mol% to 30 mol%, 23 mol% to 30 mol%, 25 mol% to 30 mol%, 26 mol% to 30 mol%, 26.5 mol% to 30 mol%, 27 mol% to 30 mol%, 27.5 mol% to 30 mol%, 28 mol% to 30 mol%, 28.5 mol% to 30 mol%, 29 mol% to 30 mol%, or 29.5 mol% to 30 mol%.

[0249] In some embodiments, the structural lipid (e.g., cholesterol) in the above- described LNPs comprises 15 mol% to 50 mol% of the total lipid present in the LNP.

[0250] In some embodiments, the structural lipid (e.g., cholesterol) in the above- described LNPs comprises 27 mol% to 42 mol% of the total lipid present in the LNP.

[0251] In some embodiments, the structural lipid (e.g., cholesterol) in the above- described LNPs comprises 30 mol% to 40 mol% of the total lipid present in the LNP.

[0252] In some embodiments, the above-described LNPs comprise an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., cholesterol), and a polymeric lipid (e.g., a PEG lipid), and do not contain stearic acid (SA) and / or its analogs and derivatives.

[0253] In some embodiments, the total lipid present in the above-described LNPs consists of an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., cholesterol), and a polymeric lipid (e.g., a PEG lipid).

[0254] In some embodiments, the above LNPs comprise an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., cholesterol), and a polymeric lipid (e.g., a PEG lipid), wherein the polymeric lipid (e.g., a PEG lipid) comprises 0.0001 mol% to 0.8 mol%, 0.01 mol% to 0.8 mol%, 0.05 mol% to 0.8 mol%, 0.1 mol% to 0.8 mol%, 0.2 mol% to 0.8 mol%, 0.3 mol% to 0.8 mol%, 0.4 mol% to 0.8 mol%, 0.5 mol% to 0.8 mol%, 0.05 mol% to 0.8 mol%, 0.1 mol% to 0.8 mol%, 0.2 mol% to 0.8 mol%, 0.3 mol% to 0.8 mol%, 0.4 mol% to 0.8 mol%, 0.0001 mol% to 0.6 mol%, 0.01 mol% to 0.6 mol%, 0.05 mol% to 0.6 mol%, 0.1 mol% to 0.6 mol%, 0.2 mol% to 0.6 mol%, 0.3 mol% to 0.6 mol%, 0.4 mol% to 0.6 mol%, 0.5 mol% to 0.6 mol%, 0.0001 mol% to 0.5 mol%, 0.01 mol% to 0.5 mol%, 0.05 mol% to 0.5 mol%, 0.1 mol% to 0.5 mol%, 0.15 mol% to 0.5 mol%, 0.2 mol% to 0.5 mol%, 0.25 mol% to 0.5 mol%, 0.3 mol% to 0.5 mol%, 0.35 mol% to 0.5 mol%, 0.4 mol% to 0.5 mol%, 0.0001 mol% to 0.45 mol%, 0.01 mol% to 0.45 mol%, 0.05 mol% to 0.45 mol%, 0.1 mol% to 0.45 mol%, 0.15 mol% to 0.45 mol%, 0.2 mol% to 0.45 mol%, 0.25 mol% to 0.45 mol%, 0.3 mol% to 0.45 mol%, 0.35 mol% to 0.45 mol%, 0.4 mol% to 0.45 mol%, 0.0001 mol% to 0.3 mol%, 0.01 mol% to 0.3 mol%, 0.05 mol% to 0.3 mol%, 0.1 mol% to 0.3 mol%, 0.15 mol% to 0.3 mol%, 0.2 mol% to 0.3 mol%, 0.0001 mol% to 0.25 mol%, 0.01 mol% to 0.25 mol%, 0.05 mol% to 0.25 mol%, 0.1 mol% to 0.25 mol%, 0.15 mol% to 0.25 mol%, 0.2 mol% to 0.25 mol%, 0.0001 mol% to 0.2 mol%, 0.01 mol% to 0.2 mol%, 0.05 mol% to 0.2 mol%, 0.1 mol% to 0.2 mol%, 0.15 mol% to 0.2 mol%, 0.0001 mol% to 0.1 mol%, 0.01 mol% to 0.1 mol%, 0.05 mol% to 0.1 mol%, 0.1 mol% of the total lipid present in the LNP.0.0001 mol% to 0.2 mol%, 0.01 mol% to 0.2 mol%, 0.05 mol% to 0.2 mol%, 0.1 mol% to 0.2 mol%, 0.15 mol% to 0.2 mol%, 0.2 mol% to 0.2 mol%, 0.0001 mol% to 0.15 mol%, 0.01 mol% to 0.15 mol%, 0.05 mol% to 0.15 mol%, 0.1 mol% to 0.15 mol%, 0.0001 mol% to 0.1 mol%, 0.01 mol% to 0.1 mol%, 0.05 mol% to 0.1 mol%, 0.0001 mol% to 0.05 mol%, or 0.01 mol% to 0.05 mol%.

[0255] In some embodiments, the LNPs described above comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 30 mol% to 60 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 0 mol% to 30 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 10 mol% to 60 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises 0 mol% to 0.8 mol% of the total lipid present in the LNPs.

[0256] In some embodiments, the LNPs described above comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 30 mol% to 60 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 0 mol% to 30 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 10 mol% to 60 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises 0.0001 mol% to 0.8 mol%, 0.001 mol% to 0.8 mol%, 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.65 mol%, 0.01 mol% to 0.6 mol%, 0.01 mol% to 0.55 mol%, 0.01 mol% to 0.5 mol%, 0.01 mol% to 0.4 mol%, 0.01 mol% to 0.3 mol%, 0.01 mol% to 0.2 mol%, 0.01 mol% to 0.15 mol%, 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.65 mol%, 0.05 mol% to 0.6 mol%, 0.05 mol% to 0.55 mol%, 0.05 mol% to 0.5 mol%, 0.05 mol% to 0.4 mol%, 0.05 mol% to 0.3 mol%, 0.05 mol% to 0.2 mol%, 0.05 mol% to 0.15 mol%, 0.05 mol% to 0.1 mol%, 0.06 mol% to 0.5 mol%, 0.06 mol% to 0.4 mol%, 0.06 mol% to 0.3 mol%, 0.06 mol% to 0.2 mol%, 0.06 mol% to 0.15 mol%, 0.06 mol% to 0.1 mol%, 0.07 mol% to 0.5 mol%, 0.07 mol% to 0.4 mol%, 0.07 mol% to 0.3 mol%, 0.07 mol% to 0.2 mol%, 0.07 mol% to 0.15 mol%, 0.07 mol% to 0.1 mol%, 0.08 mol% to 0.5 mol%, 0.08 mol% to 0.4 mol%, 0.08 mol% to 0.3 mol%, 0.08 mol% to 0.2 mol%, 0.08 mol% to 0.15 mol%, 0.08 mol% to 0.1 mol%, 0.09 mol% to 0.5 mol%, 0.09 mol% to 0.4 mol%, 0.09 mol% to 0.3 mol%, 0.09 mol% to 0.2 mol%, 0.09 mol% to 0.15 mol%, 0.09 mol% to 0.1 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.4 mol%, 0.1 mol% to 0.3 mol%, 0.1 mol% to 0.2 mol%, 0.1 mol% to 0.15 mol%, 0.1 mol% to 0.1 mol%, 0.15 mol% to 0.5 mol%, 0.15 mol% to 0.4 mol%, 0.15 mol% to 0.3 mol%, 0.15 mol% to 0.2 mol%, 0.2 mol% to 0.5 mol%, 0.2 mol% to 0.4 mol%, 0.2 mol% to 0.3 mol%, 0.3 mol% to 0.5 mol%, 0.3 mol% to 0.4 mol%, or 0.4 mol% to 0.5 mol% of the total lipid present in the LNPs.0.09 mol% to 0.2 mol%, 0.09 mol% to 0.15 mol%, 0.09 mol% to 0.1 mol%, 0.1 mol% to 0.8 mol%, 0.1 mol% to 0.8 mol%, 0.1 mol% to 0.65 mol%, 0.1 mol% to 0.6 mol%, 0.1 mol% to 0.55 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.4 mol%, 0.15 mol% to 0.8 mol%, 0.15 mol% to 0.65 mol%, 0.15 mol% to 0.6 mol%, 0.15 mol% to 0.55 mol%, 0.15 mol% to 0.5 mol%, 0.15 mol% to 0.4 mol%, 0.15 mol% to 0.8 mol%, 0.2 mol% to 0.65 mol%, 0.2 mol% to 0.6 mol%, 0.2 mol% to 0.55 mol%, 0.2 mol% to 0.5 mol%, or 0.2 mol% to 0.4 mol%.

[0257] In some embodiments, the LNPs described above comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 30 mol% to 60 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 0 mol% to 30 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 10 mol% to 60 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises 0.0001 mol% to 0.8 mol%, 0.001 mol% to 0.8 mol%, 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.65 mol%, 0.01 mol% to 0.6 mol%, 0.01 mol% to 0.55 mol%, 0.01 mol% to 0.5 mol%, 0.01 mol% to 0.4 mol%, 0.01 mol% to 0.3 mol%, 0.01 mol% to 0.2 mol%, 0.01 mol% to 0.15 mol%, 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.65 mol%, 0.05 mol% to 0.6 mol%, 0.05 mol% to 0.55 mol%, 0.05 mol% to 0.5 mol%, 0.05 mol% to 0.4 mol%, 0.05 mol% to 0.3 mol%, 0.05 mol% to 0.2 mol%, 0.05 mol% to 0.1 mol%, 0.06 mol% to 0.5 mol%, 0.05 mol% to 0.3 mol%, 0.05 mol% to 0.2 mol%, 0.05 mol% to 0.15 mol%, 0.05 mol% to 0.1 mol%, 0.06 mol% to 0.5 mol%, 0.06 mol% to 0.4 mol%, 0.06 mol% to 0.3 mol%, 0.06 mol% to 0.2 mol%, 0.06 mol% to 0.15 mol%, 0.06 mol% to 0.1 mol%, 0.07 mol% to 0.5 mol%, 0.07 mol% to 0.4 mol%, 0.07 mol% to 0.3 mol%, 0.07 mol% to 0.2 mol%, 0.07 mol% to 0.15 mol%, 0.07 mol% to 0.1 mol%, 0.08 mol% to 0.5 mol%, 0.08 mol% to 0.4 mol%, 0.08 mol% to 0.3 mol%, 0.08 mol% to 0.2 mol%, 0.08 mol% to 0.15 mol%, 0.08 mol% to 0.1 mol%, 0.09 mol% to 0.5 mol%, 0.09 mol% to 0.4 mol%, 0.09 mol% to 0.3 mol%, 0.09 mol% to 0.2 mol%, 0.09 mol% to 0.15 mol%, 0.09 mol% to 0.1 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.4 mol%, 0.1 mol% to 0.3 mol%, 0.1 mol% to 0.2 mol%, 0.1 mol% to 0.15 mol%, 0.1 mol% to 0.1 mol%, 0.1 mol% to 0.05 mol%, 0.1 mol% to 0.01 mol%, 0.1 mol% to 0.001 mol%, 0.1 mol% to 0.0001 mol%, 0.1 mol% to 0.00001 mol%, 0.1 mol% to 0.000001 mol%, 0.1 mol% to 0.0000001 mol%, 0.1 mol% to 0.00000001 mol%, 0.1 mol% to 0.000000001 mol%, 0.1 mol% to 0.0000000001 mol%, 0.1 mol% to 0.00000000001 mol%, 0.1 mol% to 0.000000000001 mol%, 0.1 mol% to 0.0000000000001 mol%, 0.1 mol% to 0.00000000000001 mol%, 0.1 mol% to 0.000000000000001 mol%, 0.1 mol% to 0.0000000000000001 mol%, 0.1 mol% to 0.00000000000000001 mol%, 0.1 mol% to 0.000000000000000001 mol%, 0.1 mol% to 0.0000000000000000001 mol%, 0.1 mol% to 0.00000000000000000001 mol%, 0.1 mol% to 0.000000000000000000001 mol%, 0.1 mol% to 0.0000000000000000000001 mol%, 0.1 mol% to 0.00000000000000000000001 mol%, 0.1 mol% to 0.000000000000000000000001 mol%, 0.1 mol% to 0.0000000000000000000000001 mol%, 0.1 mol% to 0.00000000000000000000000001 mol%, 0.1 mol% to 0.000000000000000000000000001 mol%, 0.1 mol% to 0.0000000000000000000000000001 mol%, 0.1 mol% to 0.00000000000000000000000000001 mol%, 0.1 mol% to 0.000000000000000000000000000001 mol%, 0.1 mol% to 0.0000000000000000000000000000001 mol%, 0.1 mol% to 0.00000000000000000000000000000001 mol%, 0.1 mol% to 0.000000000000000000000000000000001 mol%, 0.1 mol% to 0.0000000000000000000000000000000001 mol%, 0.1 mol% to 0.00000000000000000000000000000000001 mol%, 0.1 mol% to 0.000000000000000000000000000000000001 mol%, 0.1 mol% to 0.0000000000000000000000000000000000001 mol%, 0.1 mol% to 0.00000000000000000000000000000000000001 mol%, 0.1 mol% to 0.000000000000000000000000000000000000001 mol%, 0.1 mol% to 0.0000000000000000000000000000000000000001 mol%, 0.1 mol% to 0.00000000000000000000000000000000000000001 mol%, 0.1 mol% to 0.0000000000000000000000000000000000000000001 mol%, 0.1 mol% to 0.00000000000000000000000000000000000000000001 mol%, 0.1 mol% to 0.0000000000000000000000000000000000000000000001 mol%, 0.1 mol% to 0.00000000000000000000000000000000000000000000001 mol%, 0.1 mol% to 0.0000000000000000000000000000000000000000000000001 mol%, 0.1 mol% to 0.00000000000000000000000000000000000000000000000001 mol%, 0.1 mol% to 0.0000000000000000000000000000000000000000000000000001 mol%, 0.1 mol% to 0.00000000000000000000000000000000000000000000000000001 mol%, 0.1 mol% to 0.0000000000000000000000000000000000000000000000000000001 mol%, 0.1 mol% to 0.00000000000000000000000000000000000000000000000000000001 mol%, 0.1 mol% to 0.000000000000000000000000000000000000000000000000000000001 mol%, 0.1 mol% to 0.0000.01 mol% to 0.8 mol%, 0.01 mol% to 0.65 mol%, 0.01 mol% to 0.6 mol%, 0.01 mol% to 0.55 mol%, 0.01 mol% to 0.5 mol%, 0.01 mol% to 0.4 mol%, 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.65 mol%, 0.05 mol% to 0.6 mol%, 0.05 mol% to 0.55 mol%, 0.05 mol% to 0.5 mol%, 0.05 mol% to 0.4 mol%, 0.07 mol% to 0.8 mol%, 0.07 mol% to 0.65 mol%, 0.07 mol% to 0.6 mol%, 0.07 mol% to 0.55 mol%, 0.07 mol% to 0.5 mol%, 0.07 mol% to 0.4 mol%, 0.1 mol% to 0.8 mol%, 0.1 mol% to 0.65 mol%, 0.1 mol% to 0.6 mol%, 0.1 mol% to 0.55 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.4 mol%, 0.15 mol% to 0.8 mol%, 0.15 mol% to 0.65 mol%, 0.15 mol% to 0.6 mol%, 0.15 mol% to 0.55 mol%, 0.15 mol% to 0.5 mol%, 0.15 mol% to 0.4 mol%, 0.2 mol% to 0.65 mol%, 0.2 mol% to 0.6 mol%, 0.2 mol% to 0.55 mol%, 0.2 mol% to 0.5 mol%, or 0.2 mol% to 0.4 mol%, and suitably not 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, 0.7 mol%, 0.75 mol%, and 0.8 mol%. For example, the above LNPs comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 30 mol% to 60 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 0 mol% to 30 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 10 mol% to 60 mol% of the total lipid present in the LNPs, the polymeric lipid (e.g., a PEG lipid) comprises 0.1 mol% to 0.6 mol% of the total lipid present in the LNPs, and not 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, and 0.6 mol%.

[0258] In some embodiments, the LNPs described above comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises from 45 mol% to 55 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises from 8 mol% to 25 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises from 20 mol% to 45 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises from 0.0001 mol% to 0.8 mol%, 0.001 mol% to 0.8 mol%, 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.65 mol%, 0.01 mol% to 0.6 mol%, 0.01 mol% to 0.55 mol%, 0.01 mol% to 0.5 mol%, 0.01 mol% to 0.4 mol%, 0.01 mol% to 0.3 mol%, 0.01 mol% to 0.2 mol%, 0.01 mol% to 0.15 mol%, 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.65 mol%, 0.05 mol% to 0.6 mol%, 0.05 mol% to 0.55 mol%, 0.05 mol% to 0.5 mol%, 0.05 mol% to 0.4 mol%, 0.05 mol% to 0.3 mol%, 0.05 mol% to 0.2 mol%, 0.05 mol% to 0.15 mol%, 0.05 mol% to 0.1 mol%, 0.06 mol% to 0.5 mol%, 0.06 mol% to 0.4 mol%, 0.06 mol% to 0.3 mol%, 0.06 mol% to 0.2 mol%, 0.06 mol% to 0.15 mol%, 0.06 mol% to 0.1 mol%, 0.07 mol% to 0.5 mol%, 0.07 mol% to 0.4 mol%, 0.07 mol% to 0.3 mol%, 0.07 mol% to 0.2 mol%, 0.07 mol% to 0.15 mol%, 0.07 mol% to 0.1 mol%, 0.08 mol% to 0.5 mol%, 0.08 mol% to 0.4 mol%, 0.08 mol% to 0.3 mol%, 0.08 mol% to 0.2 mol%, 0.08 mol% to 0.15 mol%, 0.08 mol% to 0.1 mol%, 0.09 mol% to 0.5 mol%, 0.09 mol% to 0.4 mol%, 0.09 mol% to 0.3 mol%, 0.09 mol% to 0.2 mol%, 0.09 mol% to 0.15 mol%, 0.09 mol% to 0.1 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.4 mol%, 0.1 mol% to 0.3 mol%, 0.1 mol% to 0.2 mol%, 0.1 mol% to 0.15 mol%, 0.1 mol% to 0.1 mol%, 0.15 mol% to 0.5 mol%, 0.15 mol% to 0.4 mol%, 0.15 mol% to 0.3 mol%, 0.15 mol% to 0.2 mol%, 0.2 mol% to 0.5 mol%, 0.2 mol% to 0.4 mol%, 0.2 mol% to 0.3 mol%, 0.3 mol% to 0.5 mol%, 0.3 mol% to 0.4 mol%, or 0.4 mol% to 0.5 mol% of the total lipid present in the LNPs.0.09 mol% to 0.2 mol%, 0.09 mol% to 0.15 mol%, 0.09 mol% to 0.1 mol%, 0.1 mol% to 0.8 mol%, 0.1 mol% to 0.8 mol%, 0.1 mol% to 0.65 mol%, 0.1 mol% to 0.6 mol%, 0.1 mol% to 0.55 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.4 mol%, 0.15 mol% to 0.8 mol%, 0.15 mol% to 0.65 mol%, 0.15 mol% to 0.6 mol%, 0.15 mol% to 0.55 mol%, 0.15 mol% to 0.5 mol%, 0.15 mol% to 0.4 mol%, 0.15 mol% to 0.8 mol%, 0.2 mol% to 0.65 mol%, 0.2 mol% to 0.6 mol%, 0.2 mol% to 0.55 mol%, 0.2 mol% to 0.5 mol%, or 0.2 mol% to 0.4 mol%.

[0259] In some embodiments, the LNPs described above comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises from 45 mol% to 55 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises from 8 mol% to 25 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises from 20 mol% to 45 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises from 0.0001 mol% to 0.8 mol%, 0.001 mol% to 0.8 mol%, 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.65 mol%, 0.01 mol% to 0.6 mol%, 0.01 mol% to 0.55 mol%, 0.01 mol% to 0.5 mol%, 0.01 mol% to 0.4 mol%, 0.01 mol% to 0.3 mol%, 0.01 mol% to 0.2 mol%, 0.01 mol% to 0.15 mol%, 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.65 mol%, 0.05 mol% to 0.6 mol%, 0.05 mol% to 0.55 mol%, 0.05 mol% to 0.5 mol%, 0.05 mol% to 0.4 mol%, 0.05 mol% to 0.3 mol%, 0.05 mol% to 0.2 mol%, 0.05 mol% to 0.15 mol%, 0.05 mol% to 0.1 mol%, 0.06 mol% to 0.5 mol%, 0.06 mol% to 0.4 mol%, 0.06 mol% to 0.3 mol%, 0.06 mol% to 0.2 mol%, 0.06 mol% to 0.15 mol%, 0.06 mol% to 0.1 mol%, 0.07 mol% to 0.5 mol%, 0.07 mol% to 0.4 mol%, 0.07 mol% to 0.3 mol%, 0.07 mol% to 0.2 mol%, 0.07 mol% to 0.15 mol%, 0.07 mol% to 0.1 mol%, 0.08 mol% to 0.5 mol%, 0.08 mol% to 0.4 mol%, 0.08 mol% to 0.3 mol%, 0.08 mol% to 0.2 mol%, 0.08 mol% to 0.15 mol%, 0.08 mol% to 0.1 mol%, 0.09 mol% to 0.5 mol%, 0.09 mol% to 0.4 mol%, 0.09 mol% to 0.3 mol%, 0.09 mol% to 0.2 mol%, 0.09 mol% to 0.15 mol%, 0.09 mol% to 0.1 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.4 mol%, 0.1 mol% to 0.3 mol%, 0.1 mol% to 0.2 mol%, 0.1 mol% to 0.15 mol%, 0.1 mol% to 0.1 mol%, 0.15 mol% to 0.5 mol%, 0.15 mol% to 0.4 mol%, 0.15 mol% to 0.3 mol%, 0.15 mol% to 0.2 mol%, 0.2 mol% to 0.5 mol%, 0.2 mol% to 0.4 mol%, 0.2 mol% to 0.3 mol%, 0.3 mol% to 0.5 mol%, 0.3 mol% to 0.4 mol%, or 0.4 mol% to 0.5 mol% of the total lipid present in the LNPs.0.09 mol% to 0.2 mol%, 0.09 mol% to 0.15 mol%, 0.09 mol% to 0.1 mol%, 0.1 mol% to 0.8 mol%, 0.1 mol% to 0.8 mol%, 0.1 mol% to 0.65 mol%, 0.1 mol% to 0.6 mol%, 0.1 mol% to 0.55 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.4 mol%, 0.15 mol% to 0.8 mol%, 0.15 mol% to 0.65 mol%, 0.15 mol% to 0.6 mol%, 0.15 mol% to 0.55 mol%, 0.15 mol% to 0.5 mol%, 0.15 mol% to 0.4 mol%, 0.15 mol% to 0.8 mol%, 0.2 mol% to 0.65 mol%, 0.2 mol% to 0.6 mol%, 0.2 mol% to 0.55 mol%, 0.2 mol% to 0.5 mol%, or 0.2 mol% to 0.4 mol%, and suitably not 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, 0.7 mol%, 0.75 mol%, and 0.8 mol%. For example, the above LNP comprises: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 45 mol% to 55 mol% of the total lipid present in the LNP, the helper lipid (e.g., DSPC) comprises 8 mol% to 25 mol% of the total lipid present in the LNP, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 20 mol% to 45 mol% of the total lipid present in the LNP, the polymeric lipid (e.g., a PEG lipid) comprises 0.01 mol% to 0.6 mol% of the total lipid present in the LNP, and not 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, and 0.6 mol%.

[0260] In some embodiments, the LNPs described above comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises from 47 mol% to 55 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises from 10 mol% to 18 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises from 27 mol% to 42 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises from 0.0001 mol% to 0.8 mol%, 0.001 mol% to 0.8 mol%, 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.65 mol%, 0.01 mol% to 0.6 mol%, 0.01 mol% to 0.55 mol%, 0.01 mol% to 0.5 mol%, 0.01 mol% to 0.4 mol%, 0.01 mol% to 0.3 mol%, 0.01 mol% to 0.2 mol%, 0.01 mol% to 0.15 mol%, 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.65 mol%, 0.05 mol% to 0.6 mol%, 0.05 mol% to 0.55 mol%, 0.05 mol% to 0.5 mol%, 0.05 mol% to 0.4 mol%, 0.05 mol% to 0.3 mol%, 0.05 mol% to 0.2 mol%, 0.05 mol% to 0.15 mol%, 0.05 mol% to 0.1 mol%, 0.06 mol% to 0.5 mol%, 0.06 mol% to 0.4 mol%, 0.06 mol% to 0.3 mol%, 0.06 mol% to 0.2 mol%, 0.06 mol% to 0.15 mol%, 0.06 mol% to 0.1 mol%, 0.07 mol% to 0.5 mol%, 0.07 mol% to 0.4 mol%, 0.07 mol% to 0.3 mol%, 0.07 mol% to 0.2 mol%, 0.07 mol% to 0.15 mol%, 0.07 mol% to 0.1 mol%, 0.08 mol% to 0.5 mol%, 0.08 mol% to 0.4 mol%, 0.08 mol% to 0.3 mol%, 0.08 mol% to 0.2 mol%, 0.08 mol% to 0.15 mol%, 0.08 mol% to 0.1 mol%, 0.09 mol% to 0.5 mol%, 0.09 mol% to 0.4 mol%, 0.09 mol% to 0.3 mol%, 0.09 mol% to 0.2 mol%, 0.09 mol% to 0.15 mol%, 0.09 mol% to 0.1 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.4 mol%, 0.1 mol% to 0.3 mol%, 0.1 mol% to 0.2 mol%, 0.1 mol% to 0.15 mol%, 0.1 mol% to 0.1 mol%, 0.15 mol% to 0.5 mol%, 0.15 mol% to 0.4 mol%, 0.15 mol% to 0.3 mol%, 0.15 mol% to 0.2 mol%, 0.2 mol% to 0.5 mol%, 0.2 mol% to 0.4 mol%, 0.2 mol% to 0.3 mol%, 0.3 mol% to 0.5 mol%, 0.3 mol% to 0.4 mol%, or 0.4 mol% to 0.5 mol% of the total lipid present in the LNPs.0.09 mol% to 0.2 mol%, 0.09 mol% to 0.15 mol%, 0.09 mol% to 0.1 mol%, 0.1 mol% to 0.8 mol%, 0.1 mol% to 0.8 mol%, 0.1 mol% to 0.65 mol%, 0.1 mol% to 0.6 mol%, 0.1 mol% to 0.55 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.4 mol%, 0.15 mol% to 0.8 mol%, 0.15 mol% to 0.65 mol%, 0.15 mol% to 0.6 mol%, 0.15 mol% to 0.55 mol%, 0.15 mol% to 0.5 mol%, 0.15 mol% to 0.4 mol%, 0.15 mol% to 0.8 mol%, 0.2 mol% to 0.65 mol%, 0.2 mol% to 0.6 mol%, 0.2 mol% to 0.55 mol%, 0.2 mol% to 0.5 mol%, or 0.2 mol% to 0.4 mol%.

[0261] In some embodiments, the LNPs described above comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises from 47 mol% to 55 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises from 10 mol% to 18 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises from 27 mol% to 42 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises from 0.0001 mol% to 0.8 mol%, 0.001 mol% to 0.8 mol%, 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.65 mol%, 0.01 mol% to 0.6 mol%, 0.01 mol% to 0.55 mol%, 0.01 mol% to 0.5 mol%, 0.01 mol% to 0.4 mol%, 0.01 mol% to 0.3 mol%, 0.01 mol% to 0.2 mol%, 0.01 mol% to 0.15 mol%, 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.65 mol%, 0.05 mol% to 0.6 mol%, 0.05 mol% to 0.55 mol%, 0.05 mol% to 0.5 mol%, 0.05 mol% to 0.4 mol%, 0.05 mol% to 0.3 mol%, 0.05 mol% to 0.2 mol%, 0.05 mol% to 0.15 mol%, 0.05 mol% to 0.1 mol%, 0.06 mol% to 0.5 mol%, 0.06 mol% to 0.4 mol%, 0.06 mol% to 0.3 mol%, 0.06 mol% to 0.2 mol%, 0.06 mol% to 0.15 mol%, 0.06 mol% to 0.1 mol%, 0.07 mol% to 0.5 mol%, 0.07 mol% to 0.4 mol%, 0.07 mol% to 0.3 mol%, 0.07 mol% to 0.2 mol%, 0.07 mol% to 0.15 mol%, 0.07 mol% to 0.1 mol%, 0.08 mol% to 0.5 mol%, 0.08 mol% to 0.4 mol%, 0.08 mol% to 0.3 mol%, 0.08 mol% to 0.2 mol%, 0.08 mol% to 0.15 mol%, 0.08 mol% to 0.1 mol%, 0.09 mol% to 0.5 mol%, 0.09 mol% to 0.4 mol%, 0.09 mol% to 0.3 mol%, 0.09 mol% to 0.2 mol%, 0.09 mol% to 0.15 mol%, 0.09 mol% to 0.1 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.4 mol%, 0.1 mol% to 0.3 mol%, 0.1 mol% to 0.2 mol%, 0.1 mol% to 0.15 mol%, 0.1 mol% to 0.1 mol%, 0.15 mol% to 0.5 mol%, 0.15 mol% to 0.4 mol%, 0.15 mol% to 0.3 mol%, 0.15 mol% to 0.2 mol%, 0.2 mol% to 0.5 mol%, 0.2 mol% to 0.4 mol%, 0.2 mol% to 0.3 mol%, 0.3 mol% to 0.5 mol%, 0.3 mol% to 0.4 mol%, or 0.4 mol% to 0.5 mol% of the total lipid present in the LNPs.0.09 mol% to 0.2 mol%, 0.09 mol% to 0.15 mol%, 0.09 mol% to 0.1 mol%, 0.1 mol% to 0.8 mol%, 0.1 mol% to 0.8 mol%, 0.1 mol% to 0.65 mol%, 0.1 mol% to 0.6 mol%, 0.1 mol% to 0.55 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.4 mol%, 0.15 mol% to 0.8 mol%, 0.15 mol% to 0.65 mol%, 0.15 mol% to 0.6 mol%, 0.15 mol% to 0.55 mol%, 0.15 mol% to 0.5 mol%, 0.15 mol% to 0.4 mol%, 0.15 mol% to 0.8 mol%, 0.2 mol% to 0.65 mol%, 0.2 mol% to 0.6 mol%, 0.2 mol% to 0.55 mol%, 0.2 mol% to 0.5 mol%, or 0.2 mol% to 0.4 mol%, and suitably not 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, 0.7 mol%, 0.75 mol%, and 0.8 mol%. For example, the above LNP comprises: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 47 mol% to 55 mol% of the total lipid present in the LNP, the helper lipid (e.g., DSPC) comprises 10 mol% to 18 mol% of the total lipid present in the LNP, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 27 mol% to 42 mol% of the total lipid present in the LNP, the polymeric lipid (e.g., a PEG lipid) comprises 0.01 mol% to 0.6 mol% of the total lipid present in the LNP, and not 0.01 mol%, 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, and 0.6 mol%.

[0262] In some embodiments, the LNPs comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 30 mol% to 60 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 0 mol% to 30 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 10 mol% to 60 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.7 mol%, 0.05 mol% to 0.6 mol%, 0.1 mol% to 0.6 mol%, or 0.2 mol% to 0.5 mol% of the total lipid present in the LNPs.

[0263] In some embodiments, the LNPs comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 30 mol% to 60 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 0 mol% to 30 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 10 mol% to 60 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.7 mol%, 0.05 mol% to 0.6 mol%, 0.1 mol% to 0.6 mol%, or 0.2 mol% to 0.5 mol% of the total lipid present in the LNPs and is not 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, 0.7 mol%, 0.75 mol%, and 0.8 mol%.

[0264] In some embodiments, the LNPs comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 30 mol% to 60 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 0 mol% to 30 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 10 mol% to 60 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises 0.05 mol% to 0.8 mol% of the total lipid present in the LNPs.

[0265] In some embodiments, the LNPs comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 30 mol% to 60 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 0 mol% to 30 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 10 mol% to 60 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises 0.05 mol% to 0.8 mol% of the total lipid present in the LNPs and is not 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, 0.7 mol%, 0.75 mol%, and 0.8 mol%.

[0266] In some embodiments, the above LNP comprises: a LNP comprising: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 30 mol% to 60 mol% of the total lipid present in the LNP, the helper lipid (e.g., DSPC) comprises 0 mol% to 30 mol% of the total lipid present in the LNP, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 10 mol% to 60 mol% of the total lipid present in the LNP, and the polymeric lipid (e.g., a PEG lipid) comprises 0.1 mol% to 0.6 mol% of the total lipid present in the LNP. In some embodiments, the LNP comprises: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 30 mol% to 60 mol% of the total lipid present in the LNP, the helper lipid (e.g., DSPC) comprises 0 mol% to 30 mol% of the total lipid present in the LNP, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 10 mol% to 60 mol% of the total lipid present in the LNP, and the polymeric lipid (e.g., a PEG lipid) comprises 0.1 mol% to 0.6 mol% of the total lipid present in the LNP and is not 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, and 0.6 mol%.

[0267] In some embodiments, the LNPs comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 30 mol% to 60 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 0 mol% to 30 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 10 mol% to 60 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises 0.2 mol% to 0.5 mol% of the total lipid present in the LNPs. In some embodiments, the LNPs comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 30 mol% to 60 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 0 mol% to 30 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 10 mol% to 60 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises 0.2 mol% to 0.5 mol% and is not 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, and 0.5 mol% of the total lipid present in the LNPs.

[0268] In some embodiments, the LNPs comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 45 mol% to 55 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 8 mol% to 25 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 20 mol% to 45 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.7 mol%, 0.05 mol% to 0.6 mol%, 0.1 mol% to 0.6 mol%, or 0.2 mol% to 0.5 mol% of the total lipid present in the LNPs.

[0269] In some embodiments, the LNPs comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 45 mol% to 55 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 8 mol% to 25 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 20 mol% to 45 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.7 mol%, 0.05 mol% to 0.6 mol%, 0.1 mol% to 0.6 mol%, or 0.2 mol% to 0.5 mol% and is not 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, 0.7 mol%, 0.75 mol%, and 0.8 mol% of the total lipid present in the LNPs.

[0270] In some embodiments, the LNPs comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 45 mol% to 55 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 8 mol% to 25 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 20 mol% to 45 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises 0.05 mol% to 0.8 mol% of the total lipid present in the LNPs. In some embodiments, the LNPs comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 45 mol% to 55 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 8 mol% to 25 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 20 mol% to 45 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises 0.05 mol% to 0.8 mol% and is not 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, 0.7 mol%, 0.75 mol%, and 0.8 mol% of the total lipid present in the LNPs.

[0271] In some embodiments, the LNPs comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 45 mol% to 55 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 8 mol% to 25 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 20 mol% to 45 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises 0.1 mol% to 0.6 mol% of the total lipid present in the LNPs.

[0272] In some embodiments, the LNPs comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises between 45 mol% and 55 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises between 8 mol% and 25 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises between 20 mol% and 45 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises between 0.1 mol% and 0.6 mol% of the total lipid present in the LNPs and is not 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, and 0.6 mol%.

[0273] In some embodiments, the LNPs comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises between 45 mol% and 55 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises between 8 mol% and 25 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises between 20 mol% and 45 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises between 0.2 mol% and 0.5 mol% of the total lipid present in the LNPs. In some embodiments, the LNPs comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises between 45 mol% and 55 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises between 8 mol% and 25 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises between 20 mol% and 45 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises between 0.2 mol% and 0.5 mol% of the total lipid present in the LNPs and is not 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, and 0.5 mol%.

[0274] In some embodiments, the above LNP comprises: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 47 mol% to 55 mol% of the total lipid present in the LNP, the helper lipid (e.g., DSPC) comprises 10 mol% to 18 mol% of the total lipid present in the LNP, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 27 mol% to 42 mol% of the total lipid present in the LNP, and the polymeric lipid (e.g., a PEG lipid) comprises 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.7 mol%, 0.05 mol% to 0.6 mol%, 0.1 mol% to 0.6 mol%, or 0.2 mol% to 0.5 mol% of the total lipid present in the LNP.

[0275] In some embodiments, the above LNP comprises: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 47 mol% to 55 mol% of the total lipid present in the LNP, the helper lipid (e.g., DSPC) comprises 10 mol% to 18 mol% of the total lipid present in the LNP, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 27 mol% to 42 mol% of the total lipid present in the LNP, and the polymeric lipid (e.g., a PEG lipid) comprises 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.7 mol%, 0.05 mol% to 0.6 mol%, 0.1 mol% to 0.6 mol%, or 0.2 mol% to 0.5 mol% of the total lipid present in the LNP and is not 0.05 mol%, 0.07 mol%, 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, 0.6 mol%, 0.7 mol%, 0.75 mol%, and 0.8 mol%.

[0276] In some embodiments, the LNPs comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 47 mol% to 55 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 10 mol% to 18 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 27 mol% to 42 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises 0.1 mol% to 0.6 mol% of the total lipid present in the LNPs.

[0277] In some embodiments, the LNPs comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 47 mol% to 55 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 10 mol% to 18 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 27 mol% to 42 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises 0.1 mol% to 0.6 mol% of the total lipid present in the LNPs and is not 0.1 mol%, 0.15 mol%, 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, 0.5 mol%, and 0.6 mol%.

[0278] In some embodiments, the LNPs comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 47 mol% to 55 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 10 mol% to 18 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 27 mol% to 42 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises 0.2 mol% to 0.5 mol% of the total lipid present in the LNPs.

[0279] In some embodiments, the LNPs comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 47 mol% to 55 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 10 mol% to 18 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 27 mol% to 42 mol% of the total lipid present in the LNPs, the polymeric lipid (e.g., a PEG lipid) comprises 0.2 mol% to 0.5 mol% of the total lipid present in the LNPs and is not 0.2 mol%, 0.25 mol%, 0.3 mol%, 0.35 mol%, 0.4 mol%, 0.45 mol%, and 0.5 mol%.

[0280] In some embodiments, the particle size of the above LNP is 120 nm to 1000 nm. For example, 125 nm, 130 nm, 135 nm, 140 nm, 145 nm, 150 nm, 151 nm, 152 nm, 153 nm, 154 nm, 155 nm, 156 nm, 157 nm, 158 nm, 159 nm, 160 nm, 161 nm, 162 nm, 163 nm, 164 nm, 165 nm, 166 nm, 167 nm, 168 nm, 169 nm, 170 nm, 171 nm, 172 nm, 173 nm, 174 nm, 175 nm, 176 nm, 177 nm, 178 nm, 179 nm, 180 nm, 181 nm, 182 nm, 183 nm, 184 nm, 185 nm, 186 nm, 187 nm, 188 nm, 189 nm, 190 nm, 191 nm, 192 nm, 193 nm, 194 nm, 195 nm, 196 nm, 197 nm, 198 nm, 199 nm, 200 nm, 201 nm, 202 nm, 203 nm, 204 nm, 205 nm, 206 nm, 207 nm, 208 nm, 209 nm, 210 nm, 211 nm, 212 nm, 213 nm, 214 nm, 215 nm, 216 nm, 217 nm, 218 nm, 219 nm, 220 nm, 221 nm, 222 nm, 223 nm, 224 nm, 225 nm, 226 nm, 227 nm, 228 nm, 229 nm, 230 nm, 231 nm, 232 nm, 233 nm, 234 nm, 235 nm, 236 nm, 237 nm, 238 nm, 239 nm, 240 nm, 241 nm, 242 nm, 243 nm, 244 nm, 245 nm, 246 nm, 247 nm, 248 nm, 249 nm, 250 nm, 255 nm, 260 nm, 265 nm, 270 nm, 275 nm, 280 nm, 285 nm, 290 nm, 295 nm, 300 nm, 310 nm, 320 nm, 330 nm, 340 nm, 350 nm, 360 nm, 370 nm, 380 nm, 390 nm, 400 nm, 410 nm, 420 nm, 430 nm, 440 nm, 450 nm, 460 nm, 470 nm, 480 nm, 490 nm, 500 nm, 520 nm, 540 nm, 550 nm, 560 nm, 580 nm, 600 nm, 650 nm, 700 nm, 750 nm, 800 nm, 850 nm, 900 nm, 950 nm, or 990 nm. In this context, the particle size of the LNP can be represented by the z-average determined by dynamic light scattering.

[0281] In some embodiments, the particle size of the above LNP is 120 nm to 800 nm, 120 nm to 760 nm, 120 nm to 700 nm, 120 nm to 650 nm, 120 nm to 600 nm, 120 nm to 550 nm, 120 nm to 500 nm, 120 nm to 450 nm, 120 nm to 400 nm, 120 nm to 350 nm, 120 nm to 300 nm, 120 nm to 299.5 nm, 125 nm to 299.5 nm, 130 nm to 299.5 nm, 135 nm to 299.5 nm, 140 nm to 299.5 nm, 145 nm to 299.5 nm, 150 nm to 299.5 nm, 151 nm to 299.5 nm, 155 nm to 299.5 nm, 160 nm to 299.5 nm, 165 nm to 299.5 nm, 170 nm to 299.5 nm, 175 nm to 299.5 nm, 180 nm to 299.5 nm, 185 nm to 299.5 nm, 190 nm to 299.5 nm, 195 nm to 299.5 nm, 200 nm to 299.5 nm, 201 nm to 299.5 nm, 205 nm to 299.5 nm, 210 nm to 299.5 nm, 215 nm to 299.5 nm, 220 nm to 299.5 nm, 225 nm to 299.5 nm, 230 nm to 299.5 nm, 235 nm to 299.5 nm, 240 nm to 299.5 nm, 245 nm to 299.5 nm, 250 nm to 299.5 nm, 251 nm to 299.5 nm, 255 nm to 299.5 nm, 260 nm to 299.5 nm, 265 nm to 299.5 nm, 270 nm to 299.5 nm, 275 nm to 299.5 nm, 280 nm to 299.5 nm, 285 nm to 299.5 nm, 290 nm to 299.5 nm, 120 nm to 290 nm, 125 nm to 290 nm, 130 nm to 290 nm, 135 nm to 290 nm, 140 nm to 290 nm, 145 nm to 290 nm, 150 nm to 290 nm, 151 nm to 290 nm, 155 nm to 290 nm, 160 nm to 290 nm, 165 nm to 290 nm, 170 nm to 290 nm, 175 nm to 290 nm, 180 nm to 290 nm, 185 nm to 290 nm, 190 nm to 290 nm, 195 nm to 290 nm, 200 nm to 290 nm, 201 nm to 290 nm, 205 nm to 290 nm, 210 nm to 290 nm, 215 nm to 290 nm, 220 nm to 290 nm, 225 nm to 290 nm, 230 nm to 290 nm, 235 nm to 290 nm,240 nm to 290 nm, 245 nm to 290 nm, 250 nm to 290 nm, 251 nm to 290 nm, 255 nm to 290 nm, 260 nm to 290 nm, 265 nm to 290 nm, 270 nm to 290 nm, 275 nm to 290 nm, 280 nm to 290 nm, 120 nm to 280 nm, 125 nm to 280 nm, 130 nm to 280 nm, 135 nm to 280 nm, 140 nm to 280 nm, 145 nm to 280 nm, 150 nm to 280 nm, 151 nm to 280 nm, 155 nm to 280 nm, 160 nm to 280 nm, 165 nm to 280 nm, 170 nm to 280 nm, 175 nm to 280 nm, 180 nm to 280 nm, 185 nm to 280 nm, 190 nm to 280 nm, 195 nm to 280 nm, 200 nm to 280 nm, 201 nm to 280 nm, 205 nm to 280 nm, 210 nm to 280 nm, 215 nm to 280 nm, 220 nm to 280 nm, 225 nm to 280 nm, 230 nm to 280 nm, 235 nm to 280 nm, 240 nm to 280 nm, 245 nm to 280 nm, 250 nm to 280 nm, 251 nm to 280 nm, 255 nm to 280 nm, 260 nm to 280 nm, 265 nm to 280 nm, 270 nm to 280 nm, 120 nm to 270 nm, 125 nm to 270 nm, 130 nm to 270 nm, 135 nm to 270 nm, 140 nm to 270 nm, 145 nm to 270 nm, 150 nm to 270 nm, 151 nm to 270 nm, 155 nm to 270 nm, 160 nm to 270 nm, 165 nm to 270 nm, 170 nm to 270 nm, 175 nm to 270 nm, 180 nm to 270 nm, 185 nm to 270 nm, 190 nm to 270 nm, 195 nm to 270 nm, 200 nm to 270 nm, 201 nm to 270 nm, 205 nm to 270 nm, 210 nm to 270 nm, 215 nm to 270 nm, 220 nm to 270 nm, 225 nm to 270 nm, 230 nm to 270 nm, 235 nm to 270 nm, 240 nm to 270 nm, 245 nm to 270 nm, 250 nm to 270 nm, 251 nm to 270 nm, 255 nm to 270 nm, 260 nm to 270 nm, 265 nm to 270 nm, 120 nm to 260 nm, 125 nm to 260 nm, 130 nm to 260 nm, 135 nm to 260 nm, 140 nm to 260 nm, 145 nm to 260 nm, 150 nm to 260 nm, 151 nm to 260 nm, 155 nm to 260 nm, 160 nm to 260 nm, 165 nm to 260 nm, 170 nm to 260 nm, 175 nm to 260 nm, 180 nm to 260 nm, 185 nm to 260 nm, 190 nm to 260 nm, 195 nm to 260 nm, 200 nm to 260 nm, 201 nm to 260 nm, 205 nm to 260 nm, 210 nm to 260 nm, 215 nm to 260 nm, 220 nm to 260 nm, 225 nm to 260 nm, 230 nm to 260 nm, 235 nm to 260 nm, 240 nm to 260 nm, 245 nm to 260 nm, 250 nm to 260 nm, 251 nm to 260 nm, 255 nm to 260 nm, 260 nm to 260 nm, 260 nm to 260 nm, 260 nm to 260 nm, 260 nm to 260 nm, 260 nm to 260 nm, 260 nm to 260 nm, 260 nm to 260 nm, 260 nm to 260 nm, 260 nm to 260 nm, 260 nm to 260 nm, 260 nm to 260 nm, 260 nm to 260 nm, 260 nm to 260 nm, 260 nm to 260 nm, 260 nm to 260 nm, 260 nm to 260 nm, 260 nm to 260 nm150 nm to 260 nm, 151 nm to 260 nm, 155 nm to 260 nm, 160 nm to 260 nm, 165 nm to 260 nm, 170 nm to 260 nm, 175 nm to 260 nm, 180 nm to 260 nm, 185 nm to 260 nm, 190 nm to 260 nm, 195 nm to 260 nm, 200 nm to 260 nm, 201 nm to 260 nm, 205 nm to 260 nm, 210 nm to 260 nm, 215 nm to 260 nm, 220 nm to 260 nm, 225 nm to 260 nm, 230 nm to 260 nm, 235 nm to 260 nm, 240 nm to 260 nm, 245 nm to 260 nm, 250 nm to 260 nm, 251 nm to 260 nm, 255 nm to 260 nm, 120 nm to 250 nm, 125 nm to 250 nm, 130 nm to 250 nm, 135 nm to 250 nm, 140 nm to 250 nm, 145 nm to 250 nm, 150 nm to 250 nm, 151 nm to 250 nm, 155 nm to 250 nm, 160 nm to 250 nm, 165 nm to 250 nm, 170 nm to 250 nm, 175 nm to 250 nm, 180 nm to 250 nm, 185 nm to 250 nm, 190 nm to 250 nm, 195 nm to 250 nm, 200 nm to 250 nm, 201 nm to 250 nm, 205 nm to 250 nm, 210 nm to 250 nm, 215 nm to 250 nm, 220 nm to 250 nm, 225 nm to 250 nm, 230 nm to 250 nm, 235 nm to 250 nm, 240 nm to 250 nm, 245 nm to 250 nm, 120 nm to 245 nm, 125 nm to 245 nm, 130 nm to 245 nm, 135 nm to 245 nm, 140 nm to 245 nm, 145 nm to 245 nm, 150 nm to 245 nm, 151 nm to 245 nm, 155 nm to 245 nm, 160 nm to 245 nm, 165 nm to 245 nm, 170 nm to 245 nm, 175 nm to 245 nm, 180 nm to 245 nm, 185 nm to 245 nm, 190 nm to 245 nm, 195 nm to 245 nm, 200 nm to 245 nm, 201 nm to 245 nm, 205 nm to 245 nm, 210 nm to 245 nm, 215 nm to 245 nm, 220 nm to 245 nm, 225 nm to 245 nm, 230 nm to 245 nm, 235 nm to 245 nm, 240 nm to 245 nm, 120 nm to 240 nm, 125 nm to 240 nm, 130 nm to 240 nm, 135 nm to 240 nm, 140 nm to 240 nm, 145 nm to 240 nm, 150 nm to 240 nm, 151 nm to 240 nm, 155 nm to 240 nm, 160 nm to 240 nm, 165 nm to 240 nm, 170 nm to 240 nm, 175 nm to 240 nm, 180 nm to 240 nm, 185 nm to 240 nm, 190 nm to 240 nm, 195 nm to 240 nm, 200 nm to 240 nm, 201 nm to 240 nm, 205 nm to 240 nm, 210 nm to 240 nm, 215 nm to 240 nm, 220 nm to 240 nm, 225 nm to 240 nm, 230 nm to 240 nm, 235 nm to 240 nm, 240 nm to 240 nm, 120 nm to 235 nm, 125 nm to 235 nm, 130 nm to 235 nm, 135 nm to 235 nm, 140 nm to 235 nm, 145 nm to 235 nm, 150 nm to 235 nm, 151 nm to 235 nm, 155 nm to 235 nm, 160 nm to 235 nm, 165 nm to 235 nm, 170 nm to 235 nm, 175 nm to 235 nm, 180 nm to 235 nm, 185 nm to 235 nm, 190 nm to 235 nm, 195 nm to 235 nm, 200 nm to 235 nm135 nm to 240 nm, 140 nm to 240 nm, 145 nm to 240 nm, 150 nm to 240 nm, 151 nm to 240 nm, 155 nm to 240 nm, 160 nm to 240 nm, 165 nm to 240 nm, 170 nm to 240 nm, 175 nm to 240 nm, 180 nm to 240 nm, 185 nm to 240 nm, 190 nm to 240 nm, 195 nm to 240 nm, 200 nm to 240 nm, 201 nm to 240 nm, 205 nm to 240 nm, 210 nm to 240 nm, 215 nm to 240 nm, 220 nm to 240 nm, 225 nm to 240 nm, 230 nm to 240 nm, 235 nm to 240 nm, 120 nm to 235 nm, 125 nm to 235 nm, 130 nm to 235 nm, 135 nm to 235 nm, 140 nm to 235 nm, 145 nm to 235 nm, 150 nm to 235 nm, 151 nm to 235 nm, 155 nm to 235 nm, 160 nm to 235 nm, 165 nm to 235 nm, 170 nm to 235 nm, 175 nm to 235 nm, 180 nm to 235 nm, 185 nm to 235 nm, 190 nm to 235 nm, 195 nm to 235 nm, 200 nm to 235 nm, 201 nm to 235 nm, 205 nm to 235 nm, 210 nm to 235 nm, 215 nm to 235 nm, 220 nm to 235 nm, 225 nm to 235 nm, 230 nm to 235 nm, 120 nm to 230 nm, 125 nm to 230 nm, 130 nm to 230 nm, 135 nm to 230 nm, 140 nm to 230 nm, 145 nm to 230 nm, 150 nm to 230 nm, 151 nm to 230 nm, 155 nm to 230 nm, 160 nm to 230 nm, 165 nm to 230 nm, 170 nm to 230 nm, 175 nm to 230 nm, 180 nm to 230 nm, 185 nm to 230 nm, 190 nm to 230 nm, 195 nm to 230 nm, 200 nm to 230 nm, 201 nm to 230 nm, 205 nm to 230 nm, 210 nm to 230 nm, 215 nm to 230 nm, 220 nm to 230 nm, 225 nm to 230 nm, 120 nm to 225 nm, 125 nm to 225 nm, 130 nm to 225 nm, 135 nm to 225 nm, 140 nm to 225 nm, 145 nm to 225 nm, 150 nm to 225 nm, 151 nm to 225 nm, 155 nm to 225 nm, 160 nm to 225 nm, 165 nm to 225 nm,170-225 nm, 175-225 nm, 180-225 nm, 185-225 nm, 190-225 nm, 195-225 nm, 200-225 nm, 201-225 nm, 205-225 nm, 210-225 nm, 215-225 nm, 220-225 nm, 120-220 nm, 125-220 nm, 130-220 nm, 135-220 nm, 140-220 nm, 145-220 nm, 150-220 nm, 151-220 nm, 155-220 nm, 160-220 nm, 165-220 nm, 170-220 nm, 175-220 nm, 180-220 nm, 185-220 nm, 190-220 nm, 195-220 nm, 200-220 nm, 201-220 nm, 205-220 nm, 210-220 nm, 215-220 nm, 120-215 nm, 125-215 nm, 130-215 nm, 135-215 nm, 140-215 nm, 145-215 nm, 150-215 nm, 151-215 nm, 155-215 nm, 160-215 nm, 165-215 nm, 170-215 nm, 175-215 nm, 180-215 nm, 185-215 nm, 190-215 nm, 195-215 nm, 200-215 nm, 201-215 nm, 205-215 nm, 210-215 nm, 120-210 nm, 125-210 nm, 130-210 nm, 135-210 nm, 140-210 nm, 145-210 nm, 150-210 nm, 151-210 nm, 155-210 nm, 160-210 nm, 165-210 nm, 170-210 nm, 175-210 nm, 180-210 nm, 185-210 nm, 190-210 nm, 195-210 nm, 200-210 nm, 201-210 nm, 205-210 nm, 120-205 nm, 125-205 nm, 130-205 nm, 135-205 nm, 140-205 nm, 145-205 nm, 150-205 nm, 151-205 nm, 155-205 nm, 160-205 nm, 165-205 nm, 170-205 nm, 175-205 nm, 180-205 nm, 185-205 nm, 190-205 nm, 195-205 nm, 200-205 nm, 201-205 nm, 120-201 nm, 125-201 nm, 130-201 nm, 135-201 nm, 140-201 nm, 145-201 nm, 150-201 nm, 151-201 nm, 155-201 nm, 160-201 nm, 165-201 nm, 170-201 nm, 175-201 nm, 180-201 nm, 185-201 nm, 190-201 nm, 195-201 nm, 200-201 nm, 120-200 nm, 125-200 nm, 130-200 nm, 135-200 nm, 140-200 nm, 145-200 nm, 150-200 nm, 151-200 nm, 155-200 nm, 160-200 nm, 165-200 nm, 170-200 nm, 175-200 nm, 180-200 nm, 185-200 nm, 190-200 nm, 195-200 nm, 200-195 nm, 200-190 nm, 200-185 nm, 200-180 nm, 200-175 nm, 200-170 nm, 200-165 nm, 200-160 nm, 200-155 nm, 200-150 nm, 200-145 nm, 200-140 nm, 200-135 nm, 200-130 nm, 200-125 nm, 200-120 nm, 200-200 nm, 200-201 nm, 200-202 nm, 200-203 nm, 200-204 nm, 200-205 nm, 200-206 nm, 200-207 nm, 200-208 nm, 200-209 nm, 200-210 nm, 200-211 nm, 200-212 nm, 200-213 nm, 200-214 nm, 200-215 nm, 200-216 nm, 200-217 nm, 200-218 nm, 200-219 nm, 200-220 nm, 200-155 nm to 205 nm, 160 nm to 205 nm, 165 nm to 205 nm, 170 nm to 205 nm, 175 nm to 205 nm, 180 nm to 205 nm, 185 nm to 205 nm, 190 nm to 205 nm, 195 nm to 205 nm, 200 nm to 205 nm, 120 nm to 200 nm, 125 nm to 200 nm, 130 nm to 200 nm, 135 nm to 200 nm, 140 nm to 200 nm, 145 nm to 200 nm, 150 nm to 200 nm, 151 nm to 200 nm, 155 nm to 200 nm, 160 nm to 200 nm, 165 nm to 200 nm, 170 nm to 200 nm, 175 nm to 200 nm, 180 nm to 200 nm, 185 nm to 200 nm, 190 nm to 200 nm, 195 nm to 200 nm, 120 nm to 195 nm, 125 nm to 195 nm, 130 nm to 195 nm, 135 nm to 195 nm, 140 nm to 195 nm, 145 nm to 195 nm, 150 nm to 195 nm, 151 nm to 195 nm, 155 nm to 195 nm, 160 nm to 195 nm, 165 nm to 195 nm, 170 nm to 195 nm, 175 nm to 195 nm, 180 nm to 195 nm, 185 nm to 195 nm, 190 nm to 195 nm, 120 nm to 190 nm, 125 nm to 190 nm, 130 nm to 190 nm, 135 nm to 190 nm, 140 nm to 190 nm, 145 nm to 190 nm, 150 nm to 190 nm, 151 nm to 190 nm, 155 nm to 190 nm, 160 nm to 190 nm, 165 nm to 190 nm, 170 nm to 190 nm, 175 nm to 190 nm, 180 nm to 190 nm, 185 nm to 190 nm, 120 nm to 185 nm, 125 nm to 185 nm, 130 nm to 185 nm, 135 nm to 185 nm, 140 nm to 185 nm, 145 nm to 185 nm, 150 nm to 185 nm, 151 nm to 185 nm, 155 nm to 185 nm, 160 nm to 185 nm, 165 nm to 185 nm, 170 nm to 185 nm, 175 nm to 185 nm, 180 nm to 185 nm, 120 nm to 180 nm, 125 nm to 180 nm, 130 nm to 180 nm, 135 nm to 180 nm, 140 nm to 180 nm, 145 nm to 180 nm, 150 nm to 180 nm, 151 nm to 180 nm, 155 nm to 180 nm, 160 nm to 180 nm, 165 nm to 180 nm,170 nm to 180 nm, 175 nm to 180 nm, 120 nm to 175 nm, 125 nm to 175 nm, 130 nm to 175 nm, 135 nm to 175 nm, 140 nm to 175 nm, 145 nm to 175 nm, 150 nm to 175 nm, 151 nm to 175 nm, 155 nm to 175 nm, 160 nm to 175 nm, 165 nm to 175 nm, 170 nm to 175 nm, 120 nm to 170 nm, 125 nm to 170 nm, 130 nm to 170 nm, 135 nm to 170 nm, 140 nm to 170 nm, 145 nm to 170 nm, 150 nm to 170 nm, 151 nm to 170 nm, 155 nm to 170 nm, 160 nm to 170 nm, 165 nm to 170 nm, 120 nm to 165 nm, 125 nm to 165 nm, 130 nm to 165 nm, 135 nm to 165 nm, 140 nm to 165 nm, 145 nm to 165 nm, 150 nm to 165 nm, 151 nm to 165 nm, 155 nm to 165 nm, 160 nm to 165 nm, 120 nm to 160 nm, 125 nm to 160 nm, 130 nm to 160 nm, 135 nm to 160 nm, 140 nm to 160 nm, 145 nm to 160 nm, 150 nm to 160 nm, 151 nm to 160 nm, 155 nm to 160 nm, 120 nm to 155 nm, 125 nm to 155 nm, 130 nm to 155 nm, 135 nm to 155 nm, 140 nm to 155 nm, 145 nm to 155 nm, 150 nm to 155 nm, or 151 nm to 155 nm.

[0282] In some embodiments, the particle size of the above LNP is 120 nm to 450 nm, 135 nm to 450 nm, 150 nm to 450 nm, 160 nm to 450 nm, 170 nm to 450 nm, 180 nm to 450 nm, 190 nm to 450 nm, 200 nm to 450 nm, 210 nm to 450 nm, 220 nm to 450 nm, 230 nm to 450 nm, 240 nm to 450 nm, 250 nm to 450 nm, 120 nm to 400 nm, 135 nm to 400 nm, 150 nm to 400 nm, 160 nm to 400 nm, 170 nm to 400 nm, 180 nm to 400 nm, 190 nm to 400 nm, 200 nm to 400 nm, 210 nm to 400 nm, 220 nm to 450 nm, 230 nm to 400 nm, 240 nm to 400 nm, 250 nm to 400 nm, 120 nm to 380 nm, 135 nm to 380 nm, 150 nm to 380 nm, 160 nm to 380 nm, 170 nm to 380 nm, 180 nm to 380 nm, 190 nm to 380 nm, 200 nm to 380 nm, 210 nm to 380 nm, 220 nm to 380 nm, 120 nm to 360 nm, 135 nm to 360 nm, 150 nm to 360 nm, 160 nm to 360 nm, 170 nm to 360 nm, 180 nm to 360 nm, 190 nm to 360 nm, 200 nm to 360 nm, 210 nm to 360 nm, 220 nm to 360 nm, 120 nm to 340 nm, 135 nm to 340 nm, 150 nm to 340 nm, 160 nm to 340 nm, 170 nm to 340 nm, 180 nm to 340 nm, 190 nm to 340 nm, 200 nm to 340 nm, 210 nm to 340 nm, 220 nm to 340 nm, 250 nm to 340 nm, 270 nm to 340 nm, 280 nm to 340 nm, 290 nm to 340 nm, 120 nm to 320 nm, 135 nm to 320 nm, 150 nm to 320 nm, 160 nm to 320 nm, 170 nm to 320 nm, 180 nm to 320 nm, 190 nm to 320 nm, 200 nm to 320 nm, 210 nm to 320 nm, 220 nm to 320 nm, 120 nm to 300 nm, 135 nm to 300 nm, 150 nm to 300 nm, 160 nm to 300 nm, 170 nm to 300 nm, 180 nm to 300 nm, 190 nm to 300 nm, 200 nm to 300 nm, 210 nm to 300 nm, 220 nm to 300 nm, 120 nm to 290 nm,135nm~290nm, 150nm~290nm, 160nm~290nm, 170nm~290nm, 180nm~290n m, 190nm~290nm, 200nm~290nm, 210nm~290nm, 220nm~290nm, 120nm~280 nm, 135nm~280nm, 150nm~280nm, 160nm~280nm, 170nm~280nm, 180nm~280nm, 190nm~280nm, 200nm~280nm, 210nm~280nm, 220nm~280nm, 120nm~ 270nm, 135nm~270nm, 150nm~270nm, 160nm~270nm, 170nm~270nm, 180n m~270nm, 190nm~270nm, 200nm~270nm, 210nm~270nm, 220nm~270nm, 120 nm~260nm, 135nm~260nm, 150nm~260nm, 160nm~260nm, 170nm~260nm, 180nm~260nm, 190nm~260nm, 200nm~260nm, 210nm~260nm, 220nm~260nm, 120nm~250nm, 135nm~250nm, 150nm~250nm, 160nm~250nm, 170nm~250n m, 180nm~250nm, 190nm~250nm, 200nm~250nm, 210nm~250nm, 220nm~250 nm, 120nm~240nm, 135nm~240nm, 150nm~240nm, 160nm~240nm, 170nm~240nm, 180nm~240nm, 190nm~240nm, 200nm~240nm, 210nm~240nm, 220nm~ 240nm, 120nm-230nm, 135nm-230nm, 150nm-230nm, 160nm-230nm, 170nm-230nm, 180nm-230nm, 190nm-230nm, 200nm-230nm, 210nm-230nm, 220nm-230nm, 120nm-220nm, 135nm-220nm, 150nm-220nm, 160nm-220nm, 170nm-220nm, 180nm-220nm, 190nm-220nm, 200nm-220nm, or 210nm-220nm.

[0283] In some embodiments, the particle size of the above LNP is 140 nm to 450 nm, 140 nm to 400 nm, 140 nm to 390 nm, 140 nm to 380 nm, 140 nm to 370 nm, 140 nm to 360 nm, 140 nm to 350 nm, 140 nm to 340 nm, 140 nm to 330 nm, 140 nm to 320 nm, 140 nm to 310 nm, 140 nm to 300 nm, 140 nm to 290 nm, 140 nm to 280 nm, 140 nm to 270 nm, 140 nm to 260 nm, 140 nm to 250 nm, 140 nm to 240 nm, 140 nm to 230 nm, 140 nm to 230 nm, 140 nm to 210 nm, or 140 nm to 200 nm.

[0284] In some embodiments, the particle size of the above LNP is 150 nm to 450 nm, 150 nm to 400 nm, 150 nm to 390 nm, 150 nm to 380 nm, 150 nm to 370 nm, 150 nm to 360 nm, 150 nm to 350 nm, 150 nm to 340 nm, 150 nm to 330 nm, 150 nm to 320 nm, 150 nm to 310 nm, 150 nm to 300 nm, 150 nm to 290 nm, 150 nm to 280 nm, 150 nm to 270 nm, 150 nm to 260 nm, 150 nm to 250 nm, 150 nm to 240 nm, 150 nm to 230 nm, 150 nm to 210 nm, 150 nm to 200 nm, 160 nm to 320 nm, 160 nm to 310 nm, 160 nm to 300 nm, 160 nm to 290 nm, 160 nm to 280 nm, 160 nm to 270 nm, 160 nm to 260 nm, 160 nm to 250 nm, 160 nm to 240 nm, 160 nm to 230 nm, 160 nm to 230 nm, 160 nm to 210 nm, 160 nm to 200 nm, 180 nm to 320 nm, 180 nm to 310 nm, 180 nm to 300 nm, 180 nm to 290 nm, 180 nm to 280 nm, 180 nm to 270 nm, 180 nm to 260 nm, 180 nm to 250 nm, 180 nm to 240 nm, 180 nm to 230 nm, 180 nm to 230 nm, 180 nm to 210 nm, 180 nm to 200 nm, 190 nm to 320 nm, 190 nm to 310 nm, 190 nm to 300 nm, 190 nm to 290 nm, 190 nm to 280 nm, 190 nm to 270 nm, 190 nm to 260 nm, 190 nm to 250 nm, 190 nm to 240 nm, 190 nm to 230 nm, 190 nm to 230 nm, 190 nm to 210 nm, 190 nm to 200 nm, 200 nm to 320 nm, 200 nm to 310 nm, 200 nm to 300 nm, 200 nm to 290 nm, 200 nm to 280 nm, 200 nm to 270 nm, 200 nm to 260 nm, 200 nm to 250 nm, 200 nm to 240 nm, 200 nm to 230 nm, or 200 nm to 210 nm.

[0285] In some embodiments, the particle size of the above LNP is 151 nm to 229 nm, 161 nm to 229 nm, 171 nm to 229 nm, 181 nm to 229 nm, 191 nm to 229 nm, 201 nm to 229 nm, 211 nm to 229 nm, 221 nm to 229 nm, 151 nm to 219 nm, 161 nm to 219 nm, 171 nm to 219 nm, 181 nm to 219 nm, 191 nm to 219 nm, 201 nm to 219 nm, 211 nm to 219 nm, 151 nm to 209 nm, 161 nm to 209 nm, 171 nm to 209 nm, 181 nm to 209 nm, 191 nm to 209 nm, 201 nm to 209 nm, 151 nm to 199 nm, 161 nm to 199 nm, 171 nm to 199 nm, 181 nm to 199 nm, or 191 nm to 199 nm.

[0286] In some embodiments, the particle size of the above LNP is 135 nm to 420 nm. In some embodiments, the particle size of the above LNP is 135 nm to 400 nm. In some embodiments, the particle size of the above LNP is 135 nm to 380 nm. In some embodiments, the particle size of the above LNP is 135 nm to 350 nm. In some embodiments, the particle size of the above LNP is 135 nm to 320 nm. In some embodiments, the particle size of the above LNP is 135 nm to 300 nm. In some embodiments, the particle size of the above LNP is 135 nm to 290 nm. In some embodiments, the particle size of the above LNP is 135 nm to 280 nm. In some embodiments, the particle size of the above LNP is 135 nm to 270 nm. In some embodiments, the particle size of the above LNP is 135 nm to 260 nm. In some embodiments, the particle size of the above LNP is 135 nm to 250 nm. In some embodiments, the particle size of the above LNP is 135 nm to 240 nm. In some embodiments, the particle size of the above LNP is 135 nm to 230 nm. In some embodiments, the particle size of the above LNP is 135 nm to 220 nm. In some embodiments, the particle size of the above LNP is 135 nm to 200 nm. In some embodiments, the particle size of the above LNP is 135 nm to 190 nm. In some embodiments, the particle size of the above LNP is 135 nm to 180 nm.

[0287] In some embodiments, the LNPs described above have a particle size of 150 nm to 420 nm. In some embodiments, the LNPs described above have a particle size of 150 nm to 400 nm. In some embodiments, the LNPs described above have a particle size of 150 nm to 380 nm. In some embodiments, the LNPs described above have a particle size of 150 nm to 350 nm. In some embodiments, the LNPs described above have a particle size of 150 nm to 320 nm. In some embodiments, the LNPs described above have a particle size of 150 nm to 300 nm. In some embodiments, the LNPs described above have a particle size of 150 nm to 290 nm. In some embodiments, the LNPs described above have a particle size of 150 nm to 280 nm. In some embodiments, the LNPs described above have a particle size of 150 nm to 270 nm. In some embodiments, the LNPs described above have a particle size of 150 nm to 260 nm. In some embodiments, the LNPs described above have a particle size of 150 nm to 250 nm. In some embodiments, the LNPs described above have a particle size of 150 nm to 240 nm. In some embodiments, the LNPs described above have a particle size of 150 nm to 230 nm. In some embodiments, the LNPs described above have a particle size of 150 nm to 220 nm. In some embodiments, the LNPs described above have a particle size of 150 nm to 210 nm. In some embodiments, the LNPs described above have a particle size of 150 nm to 200 nm. In some embodiments, the LNPs described above have a particle size of 150 nm to 190 nm.

[0288] In some embodiments, the LNPs described above have a particle size of 160 nm to 420 nm. In some embodiments, the LNPs described above have a particle size of 160 nm to 400 nm. In some embodiments, the LNPs described above have a particle size of 160 nm to 380 nm. In some embodiments, the LNPs described above have a particle size of 160 nm to 350 nm. In some embodiments, the LNPs described above have a particle size of 160 nm to 320 nm. In some embodiments, the LNPs described above have a particle size of 160 nm to 300 nm. In some embodiments, the LNPs described above have a particle size of 160 nm to 290 nm. In some embodiments, the LNPs described above have a particle size of 160 nm to 280 nm. In some embodiments, the LNPs described above have a particle size of 160 nm to 270 nm. In some embodiments, the LNPs described above have a particle size of 160 nm to 260 nm. In some embodiments, the LNPs described above have a particle size of 160 nm to 250 nm. In some embodiments, the LNPs described above have a particle size of 160 nm to 240 nm. In some embodiments, the LNPs described above have a particle size of 160 nm to 230 nm. In some embodiments, the LNPs described above have a particle size of 160 nm to 220 nm. In some embodiments, the LNPs described above have a particle size of 160 nm to 210 nm. In some embodiments, the LNPs described above have a particle size of 160 nm to 200 nm. In some embodiments, the LNPs described above have a particle size of 160 nm to 190 nm.

[0289] In some embodiments, the particle size of the above LNP is 170 nm to 420 nm. In some embodiments, the particle size of the above LNP is 170 nm to 400 nm. In some embodiments, the particle size of the above LNP is 170 nm to 380 nm. In some embodiments, the particle size of the above LNP is 170 nm to 350 nm. In some embodiments, the particle size of the above LNP is 170 nm to 320 nm. In some embodiments, the particle size of the above LNP is 170 nm to 300 nm. In some embodiments, the particle size of the above LNP is 170 nm to 280 nm. In some embodiments, the particle size of the above LNP is 170 nm to 260 nm. In some embodiments, the particle size of the above LNP is 170 nm to 250 nm. In some embodiments, the particle size of the above LNP is 170 nm to 240 nm. In some embodiments, the particle size of the above LNP is 170 nm to 230 nm. In some embodiments, the particle size of the above LNP is 170 nm to 220 nm. In some embodiments, the particle size of the above LNP is 170 nm to 220 nm. In some embodiments, the particle size of the above LNP is 170 nm to 210 nm. In some embodiments, the particle size of the above LNP is 170 nm to 200 nm. In some embodiments, the particle size of the above LNP is 170 nm to 190 nm.

[0290] In some embodiments, the particle size of the above LNP is 180 nm to 420 nm. In some embodiments, the particle size of the above LNP is 180 nm to 400 nm. In some embodiments, the particle size of the above LNP is 180 nm to 380 nm. In some embodiments, the particle size of the above LNP is 180 nm to 350 nm. In some embodiments, the particle size of the above LNP is 180 nm to 320 nm. In some embodiments, the particle size of the above LNP is 180 nm to 300 nm. In some embodiments, the particle size of the above LNP is 180 nm to 280 nm. In some embodiments, the particle size of the above LNP is 180 nm to 260 nm. In some embodiments, the particle size of the above LNP is 180 nm to 250 nm. In some embodiments, the particle size of the above LNP is 180 nm to 240 nm. In some embodiments, the particle size of the above LNP is 180 nm to 230 nm. In some embodiments, the particle size of the above LNP is 180 nm to 220 nm. In some embodiments, the particle size of the above LNP is 180 nm to 220 nm. In some embodiments, the particle size of the above LNP is 180 nm to 210 nm. In some embodiments, the particle size of the above LNP is 180 nm to 200 nm. In some embodiments, the particle size of the above LNP is 180 nm to 190 nm.

[0291] In some embodiments, the particle size of the above-described LNPs is not one or more of: 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, 300 nm, 350 nm, 400 nm, 450 nm, 500 nm, 550 nm, 600 nm, 650 nm, 700 nm, 750 nm, 800 nm, 850 nm, 900 nm, 950 nm, and 1000 nm. For example, in some embodiments, the particle size of the above-described LNPs is 120 nm to 500 nm and is not one or more of: 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, 300 nm, 350 nm, 400 nm, 450 nm, and 500 nm. For example, in some embodiments, the particle size of the above-described LNPs is 120 nm to 300 nm and is not one or more of: 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, and 300 nm. For example, in some embodiments, the particle size of the above-described LNPs is 120 nm to 280 nm and is not one or more of: 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, and 275 nm. For example, in some embodiments, the particle size of the above-described LNPs is 150 nm to 250 nm and is not one or more of: 150 nm, 175 nm, 200 nm, 225 nm, and 250 nm. For example, in some embodiments, the particle size of the above-described LNPs is 150 nm to 230 nm and is not one or more of: 150 nm, 175 nm, 200 nm, and 225 nm.

[0292] In some embodiments, the particle size of the above-described LNPs is not one or more of: 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, 300 nm, 350 nm, 400 nm, 450 nm, 500 nm, 550 nm, 600 nm, 650 nm, 700 nm, 750 nm, 800 nm, 850 nm, 900 nm, 950 nm, and 1000 nm.

[0293] In some embodiments, the particle size of the above LNP is 120 nm to 300 nm and is not 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, and 300 nm. In some embodiments, the particle size of the above LNP is 120 nm to 290 nm and is not 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, and 275 nm. In some embodiments, the particle size of the above LNP is 120 nm to 280 nm and is not 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, and 275 nm. In some embodiments, the particle size of the above LNP is 120 nm to 270 nm and is not 150 nm, 175 nm, 200 nm, 225 nm, and 250 nm. In some embodiments, the particle size of the above LNP is 120 nm to 260 nm and is not 150 nm, 175 nm, 200 nm, 225 nm, and 250 nm. In some embodiments, the particle size of the above LNP is 120 nm to 250 nm and is not 150 nm, 175 nm, 200 nm, 225 nm, and 250 nm. In some embodiments, the particle size of the above LNP is 120 nm to 240 nm and is not 150 nm, 175 nm, 200 nm, and 225 nm. In some embodiments, the particle size of the above LNP is 120 nm to 230 nm and is not 150 nm, 175 nm, 200 nm, and 225 nm. In some embodiments, the particle size of the above LNP is 120 nm to 220 nm and is not 150 nm, 175 nm, and 200 nm. In some embodiments, the particle size of the above LNP is 120 nm to 210 nm and is not 150 nm, 175 nm, and 200 nm. In some embodiments, the particle size of the above LNP is 120 nm to 200 nm and is not 150 nm, 175 nm, and 200 nm. In some embodiments, the particle size of the above LNP is 120 nm to 190 nm and is not 150 nm and 175 nm. In some embodiments, the particle size of the above LNP is 120 nm to 180 nm and is not 150 nm and 175 nm.

[0294] In some embodiments, the LNPs described above have a particle size of 135 nm to 300 nm and not 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, and 300 nm. In some embodiments, the LNPs described above have a particle size of 135 nm to 290 nm and not 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, and 275 nm. In some embodiments, the LNPs described above have a particle size of 135 nm to 280 nm and not 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, and 275 nm. In some embodiments, the LNPs described above have a particle size of 135 nm to 270 nm and not 150 nm, 175 nm, 200 nm, 225 nm, and 250 nm. In some embodiments, the LNPs described above have a particle size of 135 nm to 260 nm and not 150 nm, 175 nm, 200 nm, 225 nm, and 250 nm. In some embodiments, the LNPs described above have a particle size of 135 nm to 250 nm and not 150 nm, 175 nm, 200 nm, 225 nm, and 250 nm. In some embodiments, the LNPs described above have a particle size of 135 nm to 240 nm and not 150 nm, 175 nm, 200 nm, and 225 nm. In some embodiments, the LNPs described above have a particle size of 135 nm to 230 nm and not 150 nm, 175 nm, 200 nm, and 225 nm. In some embodiments, the LNPs described above have a particle size of 135 nm to 220 nm and not 150 nm, 175 nm, and 200 nm. In some embodiments, the LNPs described above have a particle size of 135 nm to 200 nm and not 150 nm, 175 nm, and 200 nm. In some embodiments, the LNPs described above have a particle size of 135 nm to 190 nm and not 150 nm and 175 nm. In some embodiments, the LNPs described above have a particle size of 135 nm to 180 nm and not 150 nm and 175 nm.

[0295] In some embodiments, the particle size of the above LNP is 140 nm to 500 nm and is not 140 nm, 145 nm, 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, 300 nm, 350 nm, 400 nm, 450 nm, and 500 nm. In some embodiments, the particle size of the above LNP is 140 nm to 450 nm and is not 140 nm, 145 nm, 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, 300 nm, 350 nm, 400 nm, and 450 nm. In some embodiments, the particle size of the above LNP is 140 nm to 400 nm and is not 140 nm, 145 nm, 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, 300 nm, 350 nm, and 400 nm. In some embodiments, the particle size of the above LNP is 140 nm to 390 nm and is not 140 nm, 145 nm, 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, 300 nm, and 350 nm. In some embodiments, the particle size of the above LNP is 140 nm to 350 nm and is not 140 nm, 145 nm, 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, 300 nm, and 350 nm. In some embodiments, the particle size of the above LNP is 140 nm to 300 nm and is not 140 nm, 145 nm, 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, and 300 nm. In some embodiments, the particle size of the above LNP is 140 nm to 290 nm and is not 140 nm, 145 nm, 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, and 275 nm. In some embodiments, the particle size of the above LNP is 140 nm to 280 nm and is not 140 nm, 145 nm, 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, and 275 nm. In some embodiments, the particle size of the above LNP is 140 nm to 270 nm and is not 140 nm, 145 nm, 150 nm, 175 nm, 200 nm, 225 nm, and 250 nm. In some embodiments, the particle size of the above LNP is 140 nm to 260 nm and is not 140 nm, 145 nm, 150 nm, 175 nm, 200 nm, 225 nm, and 250 nm. In some embodiments, the particle size of the above LNP is 140 nm to 250 nm and is not 140 nm, 145 nm, 150 nm, 175 nm, 200 nm, 225 nm, and 250 nm.In some embodiments, the particle size of the above LNP is 140 nm to 240 nm and is not 140 nm, 145 nm, 150 nm, 175 nm, 200 nm, and 225 nm. In some embodiments, the particle size of the above LNP is 140 nm to 230 nm and is not 140 nm, 145 nm, 150 nm, 175 nm, 200 nm, and 225 nm. In some embodiments, the particle size of the above LNP is 140 nm to 220 nm and is not 140 nm, 145 nm, 150 nm, 175 nm, and 200 nm. In some embodiments, the particle size of the above LNP is 140 nm to 200 nm and is not 140 nm, 145 nm, 150 nm, 175 nm, and 200 nm. In some embodiments, the particle size of the above LNP is 140 nm to 190 nm and is not 140 nm, 145 nm, 150 nm, and 175 nm. In some embodiments, the particle size of the above LNP is 140 nm to 180 nm and is not 140 nm, 145 nm, 150 nm, and 175 nm.

[0296] In some embodiments, the particle size of the above LNP is 150 nm to 500 nm and is not 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, 300 nm, 350 nm, 400 nm, 450 nm, and 500 nm.

[0297] In some embodiments, the particle size of the above LNP is 150 nm to 450 nm and is not 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, 300 nm, 350 nm, 400 nm, and 450 nm. In some embodiments, the particle size of the above LNP is 150 nm to 400 nm and is not 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, 300 nm, 350 nm, and 400 nm. In some embodiments, the particle size of the above LNP is 150 nm to 350 nm and is not 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, 300 nm, and 350 nm. In some embodiments, the particle size of the above LNP is 150 nm to 320 nm and is not 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, and 300 nm. In some embodiments, the particle size of the above LNP is 150 nm to 300 nm and is not 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, and 300 nm. In some embodiments, the particle size of the above LNP is 150 nm to 290 nm and is not 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, and 275 nm. In some embodiments, the particle size of the above LNP is 150 nm to 280 nm and is not 150 nm, 175 nm, 200 nm, 225 nm, 250 nm, and 275 nm. In some embodiments, the particle size of the above LNP is 150 nm to 270 nm and is not 150 nm, 175 nm, 200 nm, 225 nm, and 250 nm. In some embodiments, the particle size of the above LNP is 150 nm to 260 nm and is not 150 nm, 175 nm, 200 nm, 225 nm, and 250 nm. In some embodiments, the particle size of the above LNP is 150 nm to 250 nm and is not 150 nm, 175 nm, 200 nm, 225 nm, and 250 nm. In some embodiments, the particle size of the above LNP is 150 nm to 240 nm and is not 150 nm, 175 nm, 200 nm, and 225 nm. In some embodiments, the particle size of the above LNP is 150 nm to 230 nm and is not 150 nm, 175 nm, 200 nm, and 225 nm. In some embodiments, the particle size of the above LNP is 150 nm to 220 nm and is not 150 nm, 175 nm, and 200 nm. In some embodiments, the particle size of the above LNP is 150 nm to 210 nm and is not 150 nm, 175 nm, and 200 nm.In some embodiments, the particle size of the above LNP is 150 nm to 200 nm and is not 150 nm, 175 nm, and 200 nm. In some embodiments, the particle size of the above LNP is 150 nm to 190 nm and is not 150 nm and 175 nm. In some embodiments, the particle size of the above LNP is 150 nm to 180 nm and is not 150 nm and 175 nm.

[0298] In some embodiments, the particle size of the above LNPs is from 160 nm to 450 nm and is not 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, 300 nm, 350 nm, 400 nm, and 450 nm. In some embodiments, the particle size of the above LNPs is from 160 nm to 400 nm and is not 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, 300 nm, 350 nm, and 400 nm. In some embodiments, the particle size of the above LNPs is from 160 nm to 350 nm and is not 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, 300 nm, and 350 nm. In some embodiments, the particle size of the above LNPs is from 160 nm to 320 nm and is not 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, and 300 nm. In some embodiments, the particle size of the above LNPs is from 160 nm to 300 nm and is not 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, and 300 nm. In some embodiments, the particle size of the above LNPs is from 160 nm to 290 nm and is not 175 nm, 200 nm, 225 nm, 250 nm, and 275 nm. In some embodiments, the particle size of the above LNPs is from 160 nm to 280 nm and is not 175 nm, 200 nm, 225 nm, 250 nm, and 275 nm. In some embodiments, the particle size of the above LNPs is from 160 nm to 270 nm and is not 175 nm, 200 nm, 225 nm, and 250 nm. In some embodiments, the particle size of the above LNPs is from 160 nm to 260 nm and is not 175 nm, 200 nm, 225 nm, and 250 nm. In some embodiments, the particle size of the above LNPs is from 160 nm to 250 nm and is not 175 nm, 200 nm, 225 nm, and 250 nm. In some embodiments, the particle size of the above LNPs is from 160 nm to 240 nm and is not 175 nm, 200 nm, and 225 nm. In some embodiments, the particle size of the above LNPs is from 160 nm to 230 nm and is not 175 nm, 200 nm, and 225 nm. In some embodiments, the particle size of the above LNPs is from 160 nm to 220 nm and is not 175 nm and 200 nm. In some embodiments, the particle size of the above LNPs is from 160 nm to 210 nm and is not 175 nm and 200 nm. In some embodiments, the particle size of the above LNPs is from 160 nm to 200 nm and is not 175 nm and 200 nm. In some embodiments, the particle size of the above LNPs is from 160 nm to 190 nm and is not 175 nm. In some embodiments, the particle size of the above LNPs is from 160 nm to 180 nm and is not 175 nm.

[0299] In some embodiments, the particle size of the above LNPs is from 170 nm to 450 nm and is not 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, 300 nm, 350 nm, 400 nm, and 450 nm. In some embodiments, the particle size of the above LNPs is from 170 nm to 400 nm and is not 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, 300 nm, 350 nm, and 400 nm. In some embodiments, the particle size of the above LNPs is from 170 nm to 350 nm and is not 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, 300 nm, and 350 nm. In some embodiments, the particle size of the above LNPs is from 170 nm to 320 nm and is not 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, and 300 nm. In some embodiments, the particle size of the above LNPs is from 170 nm to 300 nm and is not 175 nm, 200 nm, 225 nm, 250 nm, 275 nm, and 300 nm. In some embodiments, the particle size of the above LNPs is from 170 nm to 290 nm and is not 175 nm, 200 nm, 225 nm, 250 nm, and 275 nm. In some embodiments, the particle size of the above LNPs is from 170 nm to 280 nm and is not 175 nm, 200 nm, 225 nm, 250 nm, and 275 nm. In some embodiments, the particle size of the above LNPs is from 170 nm to 270 nm and is not 175 nm, 200 nm, 225 nm, and 250 nm. In some embodiments, the particle size of the above LNPs is from 170 nm to 260 nm and is not 175 nm, 200 nm, 225 nm, and 250 nm. In some embodiments, the particle size of the above LNPs is from 170 nm to 250 nm and is not 175 nm, 200 nm, 225 nm, and 250 nm. In some embodiments, the particle size of the above LNPs is from 170 nm to 240 nm and is not 175 nm, 200 nm, and 225 nm. In some embodiments, the particle size of the above LNPs is from 170 nm to 230 nm and is not 175 nm, 200 nm, and 225 nm. In some embodiments, the particle size of the above LNPs is from 170 nm to 220 nm and is not 175 nm and 200 nm. In some embodiments, the particle size of the above LNPs is from 170 nm to 210 nm and is not 175 nm and 200 nm. In some embodiments, the particle size of the above LNPs is from 170 nm to 200 nm and is not 175 nm and 200 nm. In some embodiments, the particle size of the above LNPs is from 170 nm to 190 nm and is not 175 nm. In some embodiments, the particle size of the above LNPs is from 170 nm to 180 nm and is not 175 nm.

[0300] In some embodiments, the particle size of the above LNP is: greater than 140 nm and less than 145 nm or greater than 145 nm and less than 150 nm.

[0301] In some embodiments, the particle size of the above LNP is: greater than 120 nm and less than 150 nm, greater than 150 nm and less than 175 nm, greater than 175 nm and less than 200 nm, greater than 200 nm and less than 225 nm, greater than 225 nm and less than 250 nm, greater than 250 nm and less than 275 nm, greater than 275 nm and less than 300 nm, greater than 300 nm and less than 350 nm, greater than 350 nm and less than 400 nm, greater than 400 nm and less than 450 nm, greater than 450 nm and less than 500 nm, greater than 500 nm and less than 550 nm, greater than 550 nm and less than 600 nm, greater than 600 nm and less than 650 nm, greater than 650 nm and less than 700 nm, greater than 700 nm and less than 750 nm, greater than 750 nm and less than 800 nm, greater than 800 nm and less than 850 nm, greater than 850 nm and less than 900 nm, or greater than 900 nm and less than 950 nm.

[0302] In some embodiments, the particle size of the above LNP is 140.2 nm to 144.9 nm or 145.5 nm to 149.9 nm.

[0303] In some embodiments, the particle size of the above LNP is: 120.5 nm to 149.9 nm, 150.5 nm to 174.9 m, 175.5 nm to 199.9 nm, 200.5 nm to 224.5 nm, 225.5 nm to 249.9 nm, 250.5 nm to 274.9 nm, or 275.5 nm to 299.9 nm.

[0304] In some embodiments, the particle size of the above LNP is: 125 nm to 149 nm, 152 nm to 174 m, 178 nm to 199 nm, 201 nm to 224 nm, 226 nm to 245 nm, 252 nm to 272 nm, or 276 nm to 299 nm.

[0305] In some embodiments, the particle size of the above LNP is: 125 nm to 145 nm, 155 nm to 170 m, 180 nm to 195 nm, 205 nm to 220 nm, 230 nm to 245 nm, 255 nm to 270 nm, or 280 nm to 295 nm.

[0306] In some embodiments, the particle size of the above LNP is greater than 140 nm and less than 145 nm.

[0307] In some embodiments, the particle size of the above LNP is greater than 145 nm and less than 150 nm.

[0308] In some embodiments, the particle size of the above LNP is greater than 120 nm and less than 150 nm.

[0309] In some embodiments, the particle size of the above LNP is greater than 150 nm and less than 175 nm.

[0310] In some embodiments, the particle size of the above LNP is greater than 175 nm and less than 200 nm.

[0311] In some embodiments, the particle size of the above LNP is greater than 200 nm and less than 225 nm.

[0312] In some embodiments, the particle size of the above LNP is greater than 225 nm and less than 250 nm.

[0313] In some embodiments, the particle size of the above LNP is greater than 250 nm and less than 275 nm.

[0314] In some embodiments, the particle size of the above LNP is greater than 275 nm and less than 300 nm.

[0315] In some embodiments, the particle size of the above LNP is greater than 300 nm and less than 350 nm.

[0316] In some embodiments, the particle size of the above LNP is greater than 350 nm and less than 400 nm.

[0317] In some embodiments, the particle size of the above LNP is greater than 400 nm and less than 450 nm.

[0318] In some embodiments, the particle size of the above LNP is greater than 141 nm to 144 nm.

[0319] In some embodiments, the particle size of the above LNP is greater than 146 nm to 149 nm.

[0320] In some embodiments, the particle size of the above LNP is greater than 151 nm to 154 nm.

[0321] In some embodiments, the particle size of the above LNP is 125 nm to 145 nm.

[0322] In some embodiments, the particle size of the above LNP is 155 nm to 170 m.

[0323] In some embodiments, the particle size of the above LNP is 180-195 nm.

[0324] In some embodiments, the particle size of the above LNP is 205-220 nm.

[0325] In some embodiments, the particle size of the above LNP is 230-245 nm.

[0326] In some embodiments, the particle size of the above LNP is 255-270 nm.

[0327] In some embodiments, the particle size of the above LNP is 280-295 nm.

[0328] In some embodiments, the particle size of the above LNP is 305-345 nm.

[0329] In some embodiments, the particle size of the above LNP is 355-395 nm.

[0330] In some embodiments, the particle size of the above LNP is 405-445 nm.

[0331] In some embodiments, the LNPs described above comprise an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., cholesterol), and a PEG lipid, wherein the polymeric lipid (e.g., PEG lipid) is present at 0 mol% to 0.8 mol%, 0.0001 mol% to 0.8 mol%, 0.01 mol% to 0.8 mol%, 0.05 mol% to 0.8 mol%, 0.1 mol% to 0.8 mol%, 0.2 mol% to 0.8 mol%, 0.3 mol% to 0.8 mol%, 0.4 mol% to 0.8 mol%, 0.5 mol% to 0.8 mol%, 0.05 mol% to 0.8 mol%, 0.1 mol% to 0.8 mol%, 0.2 mol% to 0.8 mol%, 0.3 mol% to 0.8 mol%, 0.4 mol% to 0.8 mol%, 0.0001 mol% to 0.6 mol%, 0.01 mol% to 0.6 mol%, 0.05 mol% to 0.6 mol%, 0.1 mol% to 0.6 mol%, 0.2 mol% to 0.6 mol%, 0.3 mol% to 0.6 mol%, 0.4 mol% to 0.6 mol%, 0.5 mol% to 0.6 mol%, 0.0001 mol% to 0.5 mol%, 0.01 mol% to 0.5 mol%, 0.05 mol% to 0.5 mol%, 0.06 mol% to 0.5 mol%, 0.06 mol% to 0.4 mol%, 0.06 mol% to 0.3 mol%, 0.06 mol% to 0.2 mol%, 0.06 mol% to 0.15 mol%, 0.06 mol% to 0.1 mol%, 0.07 mol% to 0.5 mol%, 0.07 mol% to 0.4 mol%, 0.07 mol% to 0.3 mol%, 0.07 mol% to 0.2 mol%, 0.07 mol% to 0.15 mol%, 0.07 mol% to 0.1 mol%, 0.08 mol% to 0.5 mol%, 0.08 mol% to 0.4 mol%, 0.08 mol% to 0.3 mol%, 0.08 mol% to 0.2 mol%, 0.08 mol% to 0.15 mol%, 0.08 mol% to 0.1 mol%, 0.09 mol% to 0.5 mol%, 0.09 mol% to 0.4 mol%, 0.09 mol% to 0.3 mol%, 0.09 mol% to 0.2 mol%, 0.09 mol% to 0.15 mol%, 0.09 mol% to 0.1 mol%, 0.1 mol% to 0.5 mol%, 0.15 mol% to 0.5 mol%, 0.2 mol% to 0.5 mol%, 0.25 mol% to 0.5 mol%, 0.3 mol% to 0.5 mol%, 0.35 mol% to 0.5 mol%, 0.4 mol% to 0.5 mol%, 0.45 mol% to 0.5 mol%, 0.05 mol% to 0.4 mol%, 0.1 mol% to 0.4 mol%, 0.2 mol% to 0.4 mol%, 0.25 mol% to 0.4 mol%, 0.3 mol% to 0.4 mol%, 0.35 mol% to 0.4 mol%, 0.4 mol% to 0.4 mol%, 0.45 mol% to 0.4 mol%, 0.05 mol% to 0.3 mol%, 0.1 mol% to 0.3 mol%, 0.2 mol% to 0.3 mol%, 0.25 mol% to 0.3 mol%, 0.3 mol% to 0.3 mol%, 0.35 mol% to 0.3 mol%, 0.4 mol% to 0.3 mol%, 0.45 mol% to 0.3 mol%, 0.05 mol% to 0.2 mol%, 0.1 mol% to 0.2 mol%, 0.2 mol% to 0.2 mol%, 0.25 mol% to 0.2 mol%, 0.3 mol% to 0.2 mol%, 0.35 mol% to 0.2 mol%, 0.4 mol% to 0.2 mol%, 0.45 mol% to 0.2 mol%, 0.05 mol% to 0.15 mol%, 0.1 mol% to 0.15 mol%, 0.2 mol% to 0.15 mol%, 0.25 mol% to 0.15 mol%, 0.3 mol% to 0.15 mol%, 0.35 mol% to 0.15 mol%, 0.4 mol% to 0.15 mol%, 0.45 mol% to 0.15 mol%, 0.05 mol% to 0.1 mol%, 0.1 mol% to 0.1 mol%, 0.2 mol% to 0.1 mol%, 0.25 mol% to 0.1 mol%, 0.3 mol% to 0.1 mol%, 0.35 mol% to 0.1 mol%, 0.4 mol% to 0.1 mol%, 0.45 mol% to 0.1 mol%, 0.05 mol% to 0.06 mol%, 0.1 mol% to 0.06 mol%, 0.2 mol% to 0.06 mol%, 0.25 mol% to 0.06 mol%, 0.3 mol% to 0.06 mol%, 0.35 mol% to 0.06 mol%, 0.4 mol% to 0.06 mol%, 0.45 mol% to 0.06 mol%, 0.05 mol% to 0.07 mol%, 0.1 mol% to 0.07 mol%, 0.2 mol% to 0.07 mol%, 0.25 mol% to 0.07 mol%, 0.3 mol% to 0.07 mol%, 0.35 mol% to 0.07 mol%, 0.4 mol% to 0.07 mol%, 0.45 mol% to 0.07 mol%, 0.05 mol% to 0.08 mol%, 0.1 mol% to 0.08 mol%, 0.2 mol% to 0.08 mol%, 0.25 mol% to 0.08 mol%, 0.3 mol% to 0.08 mol%, 0.35 mol% to 0.08 mol%, 0.4 mol% to 0.08 mol%, 0.45 mol% to 0.08 mol%, 0.05 mol% to 0.09 mol%, 0.1 mol% to 0.09 mol%, 0.2 mol% to 0.09 mol%, 0.25 mol% to 0.09 mol%, 0.3 mol% to 0.09 mol%, 0.35 mol% to 0.09 mol%, 0.4 mol% to 0.09 mol%, 0.45 mol% to 0.09 mol%, 0.05 mol% to 0.1 mol%, 0.1 mol% to 0.1 mol%, 0.2 mol% to 0.1 mol%, 0.25 mol% to 0.1 mol%, 0.3 mol% to 0.1 mol%, 0.35 mol% to 0.1 mol%, 0.4 mol% to 0.1 mol%, 0.45 mol% to 0.1 mol%, 0.05 mol% to 0.05 mol%, 0.1 mol% to 0.1 mol%, 0.2 mol% to 0.1 mol%, 0.25 mol% to 0.1 mol%, 0.3 mol% to 0.1 mol%, 0.35 mol% to 0.1 mol%, 0.4 mol% to 0.1 mol%, 0.45 mol% to 0.1 mol%, 0.05 mol% to 0.05 mol%, 0.1 mol% to 0.1 mol%, 0.2 mol% to 0.1 mol%, 0.25 mol% to 0.1 mol%, 0.3 mol% to 0.1 mol%, 0.35 mol% to 0.1 mol%, 0.4 mol% to 0.1 mol%, 0.45 mol% to 0.1 mol%, 00.35 mol% to 0.5 mol%, 0.4 mol% to 0.5 mol%, 0.0001 mol% to 0.45 mol%, 0.01 mol% to 0.45 mol%, 0.05 mol% to 0.45 mol%, 0.1 mol% to 0.45 mol%, 0.15 mol% to 0.45 mol%, 0.2 mol% to 0.45 mol%, 0.25 mol% to 0.45 mol%, 0.3 mol% to 0.45 mol%, 0.35 mol% to 0.45 mol%, 0.4 mol% to 0.45 mol%, 0.0001 mol% to 0.3 mol%, 0.01 mol% to 0.3 mol%, 0.05 mol% to 0.3 mol%, 0.1 mol% to 0.3 mol%, 0.15 mol% to 0.3 mol%, 0.2 mol% to 0.3 mol%, 0.0001 mol% to 0.25 mol%, 0.01 mol% to 0.25 mol%, 0.05 mol% to 0.25 mol%, 0.1 mol% to 0.25 mol%, 0.15 mol% to 0.25 mol%, 0.2 mol% to 0.25 mol%, 0.0001 mol% to 0.2 mol%, 0.01 mol% to 0.2 mol%, 0.05 mol% to 0.2 mol%, 0.1 mol% to 0.2 mol%, 0.15 mol% to 0.2 mol%, 0.2 mol% to 0.2 mol%, 0.0001 mol% to 0.15 mol%, 0.01 mol% to 0.15 mol%, 0.05 mol% to 0.15 mol%, 0.1 mol% to 0.15 mol%, 0.0001 mol% to 0.1 mol%, 0.01 mol% to 0.1 mol%, 0.05 mol% to 0.1 mol%, 0.0001 mol% to 0.05 mol%, or 0.01 mol% to 0.05 mol%, and the LNPs have a particle size of 140 nm to 500 nm, 140 nm to 450 nm, 140 nm to 400 nm, 140 nm to 390 nm, 140 nm to 380 nm, 140 nm to 370 nm, 140 nm to 360 nm, 140 nm to 350 nm, 140 nm to 340 nm, 140 nm to 330 nm, 140 nm to 320 nm, 140 nm to 310 nm, 140 nm to 300 nm, 140 nm to 290 nm, 140 nm to 280 nm, 140 nm to 270 nm, 140 nm to 260 nm, 140 nm to 250 nm, 140 nm to 240 nm, 140 nm to 230 nm, 140 nm to 210 nm, 140 nm to 200 nm, 150 nm to 500 nm, 150 nm to 450 nm, 150 nm to 400 nm,150 nm to 390 nm, 150 nm to 380 nm, 150 nm to 370 nm, 150 nm to 360 nm, 150 nm to 350 nm, 150 nm to 340 nm, 150 nm to 330 nm, 150 nm to 320 nm, 150 nm to 310 nm, 150 nm to 300 nm, 150 nm to 290 nm, 150 nm to 280 nm, 150 nm to 270 nm, 150 nm to 260 nm, 150 nm to 250 nm, 150 nm to 240 nm, 150 nm to 230 nm, 150 nm to 210 nm, 150 nm to 200 nm, 160 nm to 320 nm, 160 nm to 310 nm, 160 nm to 300 nm, 160 nm to 290 nm, 160 nm to 280 nm, 160 nm to 270 nm, 160 nm to 260 nm, 160 nm to 250 nm, 160 nm to 240 nm, 160 nm to 230 nm, 160 nm to 230 nm, 160 nm to 210 nm, 160 nm to 200 nm, 180 nm to 320 nm, 180 nm to 310 nm, 180 nm to 300 nm, 180 nm to 290 nm, 180 nm to 280 nm, 180 nm to 270 nm, 180 nm to 260 nm, 180 nm to 250 nm, 180 nm to 240 nm, 180 nm to 230 nm, 180 nm to 230 nm, 180 nm to 210 nm, 180 nm to 200 nm, 190 nm to 320 nm, 190 nm to 310 nm, 190 nm to 300 nm, 190 nm to 290 nm, 190 nm to 280 nm, 190 nm to 270 nm, 190 nm to 260 nm, 190 nm to 250 nm, 190 nm to 240 nm, 190 nm to 230 nm, 190 nm to 230 nm, 190 nm to 210 nm, 190 nm to 200 nm, 200 nm to 320 nm, 200 nm to 310 nm, 200 nm to 300 nm, 200 nm to 290 nm, 200 nm to 280 nm, 200 nm to 270 nm, 200 nm to 260 nm, 200 nm to 250 nm, 200 nm to 240 nm, 200 nm to 230 nm, or 200 nm to 210 nm.

[0332] In some embodiments, the above LNP comprises an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., cholesterol), and a PEG lipid, wherein the polymer lipid (e.g., PEG lipid) is present at 0.0001 mol% to 0.8 mol%, 0.001 mol% to 0.8 mol%, 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.7 mol%, 0.01 mol% to 0.6 mol%, 0.01 mol% to 0.5 mol%, 0.01 mol% to 0.4 mol%, 0.01 mol% to 0.3 mol%, 0.01 mol% to 0.2 mol%, 0.01 mol% to 0.1 mol%, 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.7 mol%, 0.05 mol% to 0.6 mol%, 0.05 mol% to 0.5 mol%, 0.05 mol% to 0.4 mol%, 0.05 mol% to 0.3 mol%, 0.05 mol% to 0.25 mol%, 0.05 mol% to 0.2 mol%, 0.05 mol% to 0.15 mol%, 0.05 mol% to 0.1 mol%, 0.1 mol% to 0.8 mol%, 0.1 mol% to 0.7 mol%, 0.1 mol% to 0.6 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.4 mol%, 0.1 mol% to 0.3 mol%, 0.1 mol% to 0.2 mol%, 0.1 mol% to 0.15 mol%, 0.15 mol% to 0.8 mol%, 0.15 mol% to 0.7 mol%, 0.15 mol% to 0.6 mol%, 0.15 mol% to 0.5 mol%, 0.15 mol% to 0.4 mol%, 0.15 mol% to 0.3 mol%, 0.15 mol% to 0.25 mol%, 0.2 mol% to 0.8 mol%, 0.2 mol% to 0.7 mol%, 0.2 mol% to 0.6 mol%, 0.2 mol% to 0.5 mol%, 0.2 mol% to 0.4 mol%, or 0.2 mol% to 0.35 mol% of the total lipid present in the LNP, and the LNP has a particle size of 140 nm to 500 nm, 140 nm to 400 nm, 140 nm to 390 nm, 140 nm to 380 nm, 140 nm to 370 nm, 140 nm to 360 nm, 140 nm to 350 nm, 140 nm to 340 nm, 140 nm to 330 nm, 140 nm to 320 nm, 140 nm to 310 nm, 140 nm to 300 nm, 140 nm to 290 nm, 140 nm to 280 nm, 140 nm to 270 nm, 140 nm to 260 nm, 140 nm to 250 nm, 140 nm to 240 nm, 140 nm to 230 nm, 140 nm to 220 nm, 140 nm to 210 nm, 140 nm to 200 nm, 140 nm to 190 nm, 140 nm to 180 nm, 140 nm to 170 nm, 140 nm to 160 nm, 140 nm to 150 nm, 140 nm to 160 nm, 140 nm to 170 nm, 140 nm to 180 nm, 140 nm to 190 nm, 140 nm to 200 nm, 140 nm to 210 nm, 140 nm to 220 nm, 140 nm to 230 nm, 140 nm to 240 nm, 140 nm to 250 nm, 140 nm to 260 nm, 140 nm to 270 nm, 140 nm to 280 nm, 140 nm to 290 nm, 140 nm to 300 nm, 140 nm to 310 nm, 140 nm to 320 nm, 140 nm to 330 nm, 140 nm to 340 nm, 140 nm to 350 nm, 140 nm to 360 nm, 140 nm to 370 nm, 140 nm to 380 nm, 140 nm to 390 nm, or 140 nm to 400 nm.140 nm to 230 nm, 140 nm to 210 nm, 140 nm to 200 nm, 150 nm to 450 nm, 150 nm to 400 nm, 150 nm to 390 nm, 150 nm to 380 nm, 150 nm to 370 nm, 150 nm to 360 nm, 150 nm to 350 nm, 150 nm to 340 nm, 150 nm to 330 nm, 150 nm to 320 nm, 150 nm to 310 nm, 150 nm to 300 nm, 150 nm to 290 nm, 150 nm to 280 nm, 150 nm to 270 nm, 150 nm to 260 nm, 150 nm to 250 nm, 150 nm to 240 nm, 150 nm to 230 nm, 150 nm to 210 nm, 150 nm to 200 nm, 160 nm to 320 nm, 160 nm to 310 nm, 160 nm to 300 nm, 160 nm to 290 nm, 160 nm to 280 nm, 160 nm to 270 nm, 160 nm to 260 nm, 160 nm to 250 nm, 160 nm to 240 nm, 160 nm to 230 nm, 160 nm to 230 nm, 160 nm to 210 nm, 160 nm to 200 nm, 180 nm to 320 nm, 180 nm to 310 nm, 180 nm to 300 nm, 180 nm to 290 nm, 180 nm to 280 nm, 180 nm to 270 nm, 180 nm to 260 nm, 180 nm to 250 nm, 180 nm to 240 nm, 180 nm to 230 nm, 180 nm to 230 nm, 180 nm to 210 nm, 180 nm to 200 nm, 190 nm to 320 nm, 190 nm to 310 nm, 190 nm to 300 nm, 190 nm to 290 nm, 190 nm to 280 nm, 190 nm to 270 nm, 190 nm to 260 nm, 190 nm to 250 nm, 190 nm to 240 nm, 190 nm to 230 nm, 190 nm to 230 nm, 190 nm to 210 nm, 190 nm to 200 nm, 200 nm to 320 nm, 200 nm to 310 nm, 200 nm to 300 nm, 200 nm to 290 nm, 200 nm to 280 nm, 200 nm to 270 nm, 200 nm to 260 nm, 200 nm to 250 nm, 200 nm to 240 nm, 200 nm to 230 nm, or 200 nm to 210 nm.

[0333] In some embodiments, the above LNP comprises an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., cholesterol), and a PEG lipid, wherein the polymer lipid (e.g., PEG lipid) is present at 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.6 mol%, 0.01 mol% to 0.5 mol%, 0.01 mol% to 0.3 mol%, 0.01 mol% to 0.2 mol%, 0.01 mol% to 0.15 mol%, 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.6 mol%, 0.05 mol% to 0.55 mol%, 0.05 mol% to 0.5 mol%, 0.05 mol% to 0.4 mol%, 0.05 mol% to 0.2 mol%, 0.05 mol% to 0.15 mol%, 0.1 mol% to 0.8 mol%, 0.1 mol% to 0.7 mol%, 0.1 mol% to 0.6 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.4 mol%, 0.15 mol% to 0.55 mol%, 0.15 mol% to 0.4 mol%, 0.2 mol% to 0.5 mol%, or 0.2 mol% to 0.4 mol% of the total lipid present in the LNP, and the LNP has a particle size of 140 nm to 400 nm, 140 nm to 390 nm, 140 nm to 380 nm, 140 nm to 370 nm, 140 nm to 360 nm, 140 nm to 350 nm, 140 nm to 340 nm, 140 nm to 330 nm, 140 nm to 320 nm, 140 nm to 310 nm, 140 nm to 300 nm, 140 nm to 290 nm, 140 nm to 280 nm, 140 nm to 270 nm, 140 nm to 260 nm, 140 nm to 250 nm, 140 nm to 240 nm, 140 nm to 230 nm, 140 nm to 210 nm, 140 nm to 200 nm, 150 nm to 400 nm, 150 nm to 350 nm, 150 nm to 300 nm, 150 nm to 250 nm, 150 nm to 200 nm, 160 nm to 350 nm, 160 nm to 300 nm, 160 nm to 250 nm, 160 nm to 200 nm, 180 nm to 320 nm, 180 nm to 300 nm, 180 nm to 250 nm, 180 nm to 200 nm, 190 nm to 320 nm, 190 nm to 300 nm, 190 nm to 250 nm, 190 nm to 200 nm, 200 nm to 320 nm, 200 nm to 300 nm, 200 nm to 250 nm, or 200 nm to 230 nm.

[0334] In some embodiments, the LNPs comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 30 mol% to 60 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 0 mol% to 30 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 10 mol% to 60 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises 0 mol% to 0.8 mol%, 0.0001 mol% to 0.8 mol%, 0.01 mol% to 0.8 mol%, 0.05 mol% to 0.8 mol%, 0.1 mol% to 0.8 mol%, 0.2 mol% to 0.8 mol%, 0.3 mol% to 0.8 mol%, 0.4 mol% to 0.8 mol%, 0.5 mol% to 0.8 mol%, 0.05 mol% to 0.8 mol%, 0.1 mol% to 0.8 mol%, 0.2 mol% to 0.8 mol%, 0.3 mol% to 0.8 mol%, 0.4 mol% to 0.8 mol%, 0.0001 mol% to 0.6 mol%, 0.01 mol% to 0.6 mol%, 0.05 mol% to 0.6 mol%, 0.1 mol% to 0.6 mol%, 0.2 mol% to 0.6 mol%, 0.3 mol% to 0.6 mol%, 0.4 mol% to 0.6 mol%, 0.5 mol% to 0.6 mol%, 0.0001 mol% to 0.5 mol%, 0.01 mol% to 0.5 mol%, 0.05 mol% to 0.5 mol%, 0.06 mol% to 0.5 mol%, 0.06 mol% to 0.4 mol%, 0.06 mol% to 0.3 mol%, 0.06 mol% to 0.2 mol%, 0.06 mol% to 0.15 mol%, 0.06 mol% to 0.1 mol%, 0.07 mol% to 0.5 mol%, 0.07 mol% to 0.4 mol%, 0.07 mol% to 0.3 mol%, 0.07 mol% to 0.2 mol%, 0.07 mol% to 0.15 mol%, 0.07 mol% to 0.1 mol%, 0.08 mol% to 0.5 mol%, 0.08 mol% to 0.4 mol%, 0.08 mol% to 0.3 mol%, 0.08 mol% to 0.2 mol%, 0.08 mol% to 0.15 mol%, 0.08 mol% to 0.1 mol%, 0.09 mol% to 0.5 mol%, 0.09 mol% to 0.4 mol%, 0.09 mol% to 0.3 mol%, 0.09 mol% to 0.2 mol%, 0.09 mol% to 0.15 mol%, 0.09 mol% to 0.1 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.4 mol%, 0.1 mol% to 0.3 mol%, 0.1 mol% to 0.2 mol%, 0.1 mol% to 0.15 mol%, 0.1 mol% to 0.1 mol%, 0.2 mol% to 0.5 mol%, 0.2 mol% to 0.4 mol%, 0.2 mol% to 0.3 mol%, 0.2 mol% to 0.2 mol%, 0.2 mol% to 0.15 mol%, 0.2 mol% to 0.1 mol%, 0.3 mol% to 0.5 mol%, 0.3 mol% to 0.4 mol%, 0.3 mol% to 0.3 mol%, 0.3 mol% to 0.15 mol%, 0.3 mol% to 0.1 mol%, 0.4 mol% to 0.5 mol%, 0.4 mol% to 0.4 mol%, 0.4 mol% to 0.3 mol%, 0.4 mol% to 0.15 mol%, 0.4 mol% to 0.1 mol%, 0.5 mol% to 0.5 mol%, 0.5 mol% to 0.4 mol%, 0.5 mol% to 0.3 mol%, 0.5 mol% to 0.15 mol%, or 0.5 mol% to 0.1 mol% of the total lipid present in the LNPs.0.09 mol% to 0.2 mol%, 0.09 mol% to 0.15 mol%, 0.09 mol% to 0.1 mol%, 0.1 mol% to 0.5 mol%, 0.15 mol% to 0.5 mol%, 0.2 mol% to 0.5 mol%, 0.25 mol% to 0.5 mol%, 0.3 mol% to 0.5 mol%, 0.35 mol% to 0.5 mol%, 0.4 mol% to 0.5 mol%, 0.0001 mol% to 0.45 mol%, 0.01 mol% to 0.45 mol%, 0.05 mol% to 0.45 mol%, 0.1 mol% to 0.45 mol%, 0.15 mol% to 0.45 mol%, 0.2 mol% to 0.45 mol%, 0.25 mol% to 0.45 mol%, 0.3 mol% to 0.45 mol%, 0.35 mol% to 0.45 mol%, 0.4 mol% to 0.45 mol%, 0.0001 mol% to 0.3 mol%, 0.01 mol% to 0.3 mol%, 0.05 mol% to 0.3 mol%, 0.1 mol% to 0.3 mol%, 0.15 mol% to 0.3 mol%, 0.2 mol% to 0.3 mol%, 0.0001 mol% to 0.25 mol%, 0.01 mol% to 0.25 mol%, 0.05 mol% to 0.25 mol%, 0.1 mol% to 0.25 mol%, 0.15 mol% to 0.25 mol%, 0.2 mol% to 0.25 mol%, 0.0001 mol% to 0.2 mol%, 0.01 mol% to 0.2 mol%, 0.05 mol% to 0.2 mol%, 0.1 mol% to 0.2 mol%, 0.15 mol% to 0.2 mol%, 0.2 mol% to 0.2 mol%, 0.0001 mol% to 0.15 mol%, 0.01 mol% to 0.15 mol%, 0.05 mol% to 0.15 mol%, 0.1 mol% to 0.15 mol%, 0.0001 mol% to 0.1 mol%, 0.01 mol% to 0.1 mol%, 0.05 mol% to 0.1 mol%, 0.0001 mol% to 0.05 mol%, or 0.01 mol% to 0.05 mol%, and the LNPs have a particle size of 140 nm to 500 nm, 140 nm to 450 nm, 140 nm to 400 nm, 140 nm to 390 nm, 140 nm to 380 nm, 140 nm to 370 nm, 140 nm to 360 nm, 140 nm to 350 nm, 140 nm to 340 nm, 140 nm to 330 nm, 140 nm to 320 nm, 140 nm to 310 nm, 140 nm to 300 nm, 140 nm to 290 nm, 140 nm to 280 nm, 140 nm to 270 nm, 140 nm to 260 nm, 140 nm to 250 nm, 140 nm to 240 nm, 140 nm to 230 nm, 140 nm to 220 nm, 140 nm to 210 nm, 140 nm to 200 nm, 140 nm to 190 nm, 140 nm to 180 nm, 140 nm to 170 nm, 140 nm to 160 nm, 140 nm to 150 nm, 140 nm to 160 nm, 140 nm to 170 nm, 140 nm to 180 nm, 140 nm to 190 nm, 140 nm to 200 nm, 140 nm to 210 nm, 140 nm to 220 nm, 140 nm to 230 nm, 140 nm to 240 nm, 140 nm to 250 nm, 140 nm to 260 nm, 140 nm to 270 nm, 140 nm to 280 nm, 140 nm to 290 nm, 140 nm to 300 nm, 140 nm to 310 nm, 140 nm to 320 nm, 140 nm to 330 nm, 140 nm to 340 nm, 140 nm to 350 nm, 140 nm to 360 nm, 140 nm to 370 nm, 140 nm to 380 nm, 140 nm to 390 nm, 140 nm to 400 nm, 140 nm to 410 nm, 140 nm to 420 nm, 140 nm to 430 nm, 140 nm to 440 nm, 140 nm to 450 nm, 140 nm to 460 nm, 140 nm to 470 nm, 140 nm to 480 nm, 140 nm to 490 nm, or 140 nm to 500 nm.140 nm to 280 nm, 140 nm to 270 nm, 140 nm to 260 nm, 140 nm to 250 nm, 140 nm to 240 nm, 140 nm to 230 nm, 140 nm to 210 nm, 140 nm to 200 nm, 150 nm to 500 nm, 150 nm to 450 nm, 150 nm to 400 nm, 150 nm to 390 nm, 150 nm to 380 nm, 150 nm to 370 nm, 150 nm to 360 nm, 150 nm to 350 nm, 150 nm to 340 nm, 150 nm to 330 nm, 150 nm to 320 nm, 150 nm to 310 nm, 150 nm to 300 nm, 150 nm to 290 nm, 150 nm to 280 nm, 150 nm to 270 nm, 150 nm to 260 nm, 150 nm to 250 nm, 150 nm to 240 nm, 150 nm to 230 nm, 150 nm to 210 nm, 150 nm to 200 nm, 160 nm to 320 nm, 160 nm to 310 nm, 160 nm to 300 nm, 160 nm to 290 nm, 160 nm to 280 nm, 160 nm to 270 nm, 160 nm to 260 nm, 160 nm to 250 nm, 160 nm to 240 nm, 160 nm to 230 nm, 160 nm to 230 nm, 160 nm to 210 nm, 160 nm to 200 nm, 180 nm to 320 nm, 180 nm to 310 nm, 180 nm to 300 nm, 180 nm to 290 nm, 180 nm to 280 nm, 180 nm to 270 nm, 180 nm to 260 nm, 180 nm to 250 nm, 180 nm to 240 nm, 180 nm to 230 nm, 180 nm to 230 nm, 180 nm to 210 nm, 180 nm to 200 nm, 190 nm to 320 nm, 190 nm to 310 nm, 190 nm to 300 nm, 190 nm to 290 nm, 190 nm to 280 nm, 190 nm to 270 nm, 190 nm to 260 nm, 190 nm to 250 nm, 190 nm to 240 nm, 190 nm to 230 nm, 190 nm to 230 nm, 190 nm to 210 nm, 190 nm to 200 nm, 200 nm to 320 nm, 200 nm to 310 nm, 200 nm to 300 nm, 200 nm to 290 nm, 200 nm to 280 nm, 200 nm to 270 nm, 200 nm to 260 nm, 200 nm to 250 nm, 200 nm to 240 nm, 200 nm to 230 nm, or 200 nm to 210 nm.

[0335] In some embodiments, LNPs comprise: ionizable lipids, accessory lipids (e.g., DSPC), structural lipids (e.g., non-phytosterols (e.g., cholesterol)), and polymeric lipids (e.g., PEG lipids), wherein the ionizable lipids account for 30 mol% to 60 mol% of the total lipids present in the LNPs, the accessory lipids (e.g., DSPC) account for 0 mol% to 30 mol% of the total lipids present in the LNPs, the structural lipids (e.g., non-phytosterols (e.g., cholesterol)) account for 10 mol% to 60 mol% of the total lipids present in the LNPs, and the polymeric lipids (e.g., PEG lipids) account for 0.0001 mol% to 0.8 mol% and 0.001 mol% to 0.8 mol% of the total lipids present in the LNPs, respectively. %, 0.01mol%~0.8mol%, 0.01mol%~0.7mol%, 0.01mol%~0.6mol%, 0.01mol%~0.5mol%, 0.01mol%~0.4mol%, 0.01mol%~0.3mol%, 0.01mol%~0.2mol%, 0.01 mol%~0.1mol%, 0.05mol%~0.8mol%, 0.05mol%~0.7mol%, 0.05mol%~0.6mol%, 0.05mol%~0.5mol%, 0.05mol%~0.4mol%, 0.05mol%~0.3mol%, 0.05mol%~0 .25mol%, 0.05mol%~0.2mol%, 0.05mol%~0.15mol%, 0.05mol%~0.1mol%, 0.1mol%~0.8mol%, 0.1mol%~0.7mol%, 0.1mol%~0.6mol%, 0.1mol%~0.5mol%, 0 .1mol%~0.4mol%, 0.1mol%~0.3mol%, 0.1mol%~0.2mol%, 0.1mol%~0.15mol%, 0.15mol%~0.8mol%, 0.15mol%~0.7mol%, 0.15mol%~0.6mol%, 0.15mol%~0 The particle sizes of the above LNPs are 0.5 mol%, 0.15 mol%–0.4 mol%, 0.15 mol%–0.3 mol%, 0.15 mol%–0.25 mol%, 0.2 mol%–0.8 mol%, 0.2 mol%–0.7 mol%, 0.2 mol%–0.6 mol%, 0.2 mol%–0.5 mol%, 0.2 mol%–0.4 mol%, or 0.2 mol%–0.35 mol%, with particle sizes of 140 nm–500 nm, 140 nm–450 nm, 140 nm–400 nm, 140 nm–390 nm, 140 nm–380 nm, 140 nm–370 nm, and 140 nm–360 nm.140 nm to 350 nm, 140 nm to 340 nm, 140 nm to 330 nm, 140 nm to 320 nm, 140 nm to 310 nm, 140 nm to 300 nm, 140 nm to 290 nm, 140 nm to 280 nm, 140 nm to 270 nm, 140 nm to 260 nm, 140 nm to 250 nm, 140 nm to 240 nm, 140 nm to 230 nm, 140 nm to 210 nm, 140 nm to 200 nm, 150 nm to 450 nm, 150 nm to 400 nm, 150 nm to 390 nm, 150 nm to 380 nm, 150 nm to 370 nm, 150 nm to 360 nm, 150 nm to 350 nm, 150 nm to 340 nm, 150 nm to 330 nm, 150 nm to 320 nm, 150 nm to 310 nm, 150 nm to 300 nm, 150 nm to 290 nm, 150 nm to 280 nm, 150 nm to 270 nm, 150 nm to 260 nm, 150 nm to 250 nm, 150 nm to 240 nm, 150 nm to 230 nm, 150 nm to 210 nm, 150 nm to 200 nm, 160 nm to 320 nm, 160 nm to 310 nm, 160 nm to 300 nm, 160 nm to 290 nm, 160 nm to 280 nm, 160 nm to 270 nm, 160 nm to 260 nm, 160 nm to 250 nm, 160 nm to 240 nm, 160 nm to 230 nm, 160 nm to 230 nm, 160 nm to 210 nm, 160 nm to 200 nm, 180 nm to 320 nm, 180 nm to 310 nm, 180 nm to 300 nm, 180 nm to 290 nm, 180 nm to 280 nm, 180 nm to 270 nm, 180 nm to 260 nm, 180 nm to 250 nm, 180 nm to 240 nm, 180 nm to 230 nm, 180 nm to 230 nm, 180 nm to 210 nm, 180 nm to 200 nm, 190 nm to 320 nm, 190 nm to 310 nm, 190 nm to 300 nm, 190 nm to 290 nm, 190 nm to 280 nm, 190 nm to 270 nm, 190 nm to 260 nm, 190 nm to 250 nm, 190 nm to 240 nm, 190 nm to 230 nm, 190 nm to 230 nm, 190 nm to 210 nm, 190 nm to 200 nm, 200 nm to 320 nm, 200 nm to 310 nm, 200 nm to 300 nm, 200 nm to 290 nm, 200 nm to 280 nm, 200 nm to 270 nm, 200 nm to 260 nm, 200 nm to 250 nm,200 nm to 240 nm, 200 nm to 230 nm, or 200 nm to 210 nm.

[0336] In some embodiments, the LNPs comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 30 mol% to 60 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 0 mol% to 30 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 10 mol% to 60 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.6 mol%, 0.01 mol% to 0.3 mol%, 0.01 mol% to 0.2 mol%, 0.01 mol% to 0.5 mol%, 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.6 mol%, 0.05 mol% to 0.55 mol%, 0.05 mol% to 0.5 mol%, 0.05 mol% to 0.4 mol%, 0.05 mol% to 0.2 mol%, 0.05 mol% to 0.15 mol%, 0.1 mol% to 0.8 mol%, 0.1 mol% to 0.7 mol%, 0.1 mol% to 0.6 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.4 mol%, 0.15 mol% to 0.55 mol%, 0.15 mol% to 0.4 mol%, 0.2 mol% to 0.5 mol%, or 0.2 mol% to 0.4 mol% of the total lipid present in the LNPs.4 mol%, the LNPs described above have a particle size of 140 nm to 400 nm, 140 nm to 390 nm, 140 nm to 380 nm, 140 nm to 370 nm, 140 nm to 360 nm, 140 nm to 350 nm, 140 nm to 340 nm, 140 nm to 330 nm, 140 nm to 320 nm, 140 nm to 310 nm, 140 nm to 300 nm, 140 nm to 290 nm, 140 nm to 280 nm, 140 nm to 270 nm, 140 nm to 260 nm, 140 nm to 250 nm, 140 nm to 240 nm, 140 nm to 230 nm, 140 nm to 210 nm, 140 nm to 200 nm, 150 nm to 400 nm, 150 nm to 350 nm, 150 nm to 300 nm, 150 nm to 250 nm, 150 nm to 200 nm, 160 nm to 350 nm, 160 nm to 300 nm, 160 nm to 250 nm, 160 nm to 200 nm, 180 nm to 320 nm, 180 nm to 300 nm, 180 nm to 250 nm, 180 nm to 200 nm, 190 nm to 320 nm, 190 nm to 300 nm, 190 nm to 250 nm, 190 nm to 200 nm, 200 nm to 320 nm, 200 nm to 300 nm, 200 nm to 250 nm, or 200 nm to 230 nm.

[0337] In some embodiments, the LNPs comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises from 45 mol% to 55 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises from 8 mol% to 25 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises from 20 mol% to 45 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises from 0.0001 mol% to 0.8 mol%, 0.001 mol% to 0.8 mol%, 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.7 mol%, 0.01 mol% to 0.6 mol%, 0.01 mol% to 0.5 mol%, 0.01 mol% to 0.4 mol%, 0.01 mol% to 0.3 mol%, 0.01 mol% to 0.2 mol%, 0.01 mol% to 0.1 mol%, 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.7 mol%, 0.05 mol% to 0.6 mol%, 0.05 mol% to 0.5 mol%, 0.05 mol% to 0.4 mol%, 0.05 mol% to 0.3 mol%, 0.05 mol% to 0.25 mol%, 0.05 mol% to 0.2 mol%, 0.05 mol% to 0.15 mol%, 0.05 mol% to 0.1 mol%, 0.06 mol% to 0.5 mol%, 0.06 mol% to 0.4 mol%, 0.06 mol% to 0.3 mol%, 0.06 mol% to 0.2 mol%, 0.06 mol% to 0.15 mol%, 0.06 mol% to 0.1 mol%, 0.07 mol% to 0.5 mol%, 0.07 mol% to 0.4 mol%, 0.07 mol% to 0.3 mol%, 0.07 mol% to 0.2 mol%, 0.07 mol% to 0.15 mol%, 0.07 mol% to 0.1 mol%, 0.08 mol% to 0.5 mol%, 0.08 mol% to 0.4 mol%, 0.08 mol% to 0.3 mol%, 0.08 mol% to 0.2 mol%, 0.08 mol% to 0.15 mol%, 0.08 mol% to 0.1 mol%, 0.09 mol% to 0.5 mol%, 0.09 mol% to 0.4 mol%, 0.09 mol% to 0.3 mol%, 0.09 mol% to 0.2 mol%, 0.09 mol% to 0.15 mol%, 0.09 mol% to 0.1 mol%, 0.1 mol% to 0.8 mol%, 0.1 mol% to 0.7 mol%, 0.1 mol% to 0.6 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.4 mol%, 0.1 mol% to 0.3 mol%, 0.1 mol% to 0.2 mol%, 0.1 mol% to 0.15 mol%, 0.1 mol% to 0.1 mol%, 0.1 mol% to 0.05 mol%, 0.2 mol% to 0.8 mol%, 0.2 mol% to 0.7 mol%, 0.2 mol% to 0.6 mol%, 0.2 mol% to 0.5 mol%, 0.2 mol% to 0.4 mol%, 0.2 mol% to 0.3 mol%, 0.3 mol% to 0.8 mol%, 0.3 mol% to 0.7 mol%, 0.3 mol% to 0.6 mol%, 0.3 mol% to 0.5 mol%, 0.3 mol% to 0.4 mol%, 0.4 mol% to 0.8 mol%, 0.4 mol% to 0.7 mol%, 0.4 mol% to 0.6 mol%, 0.4 mol% to 0.5 mol%, 0.5 mol% to 0.8 mol%, 0.5 mol% to 0.7 mol%, 0.5 mol% to 0.6 mol%, 0.6 mol% to 0.8 mol%, 0.6 mol% to 0.7 mol%, or 0.7 mol% to 0.8 mol% of the total lipid present in the LNPs.0.1 mol% to 0.7 mol%, 0.1 mol% to 0.6 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.4 mol%, 0.1 mol% to 0.3 mol%, 0.1 mol% to 0.2 mol%, 0.1 mol% to 0.15 mol%, 0.15 mol% to 0.8 mol%, 0.15 mol% to 0.7 mol%, 0.15 mol% to 0.6 mol%, 0.15 mol% to 0.5 mol%, 0.15 mol% to 0.4 mol%, 0.15 mol% to 0.3 mol%, 0.15 mol% to 0.25 mol%, 0.2 mol% to 0.8 mol%, 0.2 mol% to 0.7 mol%, 0.2 mol% to 0.6 mol%, 0.2 mol% to 0.5 mol%, 0.2 mol% to 0.4 mol%, or 0.2 mol% to 0.35 mol%, and the LNPs have a particle size of 140 nm to 500 nm, 140 nm to 450 nm, 140 nm to 400 nm, 140 nm to 390 nm, 140 nm to 380 nm, 140 nm to 370 nm, 140 nm to 360 nm, 140 nm to 350 nm, 140 nm to 340 nm, 140 nm to 330 nm, 140 nm to 320 nm, 140 nm to 310 nm, 140 nm to 300 nm, 140 nm to 290 nm, 140 nm to 280 nm, 140 nm to 270 nm, 140 nm to 260 nm, 140 nm to 250 nm, 140 nm to 240 nm, 140 nm to 230 nm, 140 nm to 210 nm, 140 nm to 200 nm, 150 nm to 450 nm, 150 nm to 400 nm, 150 nm to 390 nm, 150 nm to 380 nm, 150 nm to 370 nm, 150 nm to 360 nm, 150 nm to 350 nm, 150 nm to 340 nm, 150 nm to 330 nm, 150 nm to 320 nm, 150 nm to 310 nm, 150 nm to 300 nm, 150 nm to 290 nm, 150 nm to 280 nm, 150 nm to 270 nm, 150 nm to 260 nm, 150 nm to 250 nm, 150 nm to 240 nm, 150 nm to 230 nm, 150 nm to 210 nm, 150 nm to 200 nm, 160 nm to 320 nm, 160 nm to 310 nm, 160 nm to 300 nm, 160 nm to 290 nm, 160 nm to 280 nm, 160 nm to 270 nm, 160 nm to 260 nm, 160 nm to 250 nm, 160 nm to 240 nm, 160 nm to 230 nm, 160 nm to 230 nm, 160 nm to 210 nm,160 nm to 200 nm, 180 nm to 320 nm, 180 nm to 310 nm, 180 nm to 300 nm, 180 nm to 290 nm, 180 nm to 280 nm, 180 nm to 270 nm, 180 nm to 260 nm, 180 nm to 250 nm, 180 nm to 240 nm, 180 nm to 230 nm, 180 nm to 230 nm, 180 nm to 210 nm, 180 nm to 200 nm, 190 nm to 320 nm, 190 nm to 310 nm, 190 nm to 300 nm, 190 nm to 290 nm, 190 nm to 280 nm, 190 nm to 270 nm, 190 nm to 260 nm, 190 nm to 250 nm, 190 nm to 240 nm, 190 nm to 230 nm, 190 nm to 230 nm, 190 nm to 210 nm, 190 nm to 200 nm, 200 nm to 320 nm, 200 nm to 310 nm, 200 nm to 300 nm, 200 nm to 290 nm, 200 nm to 280 nm, 200 nm to 270 nm, 200 nm to 260 nm, 200 nm to 250 nm, 200 nm to 240 nm, 200 nm to 230 nm, or 200 nm to 210 nm.

[0338] In some embodiments, the LNPs comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 45 mol% to 55 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 8 mol% to 25 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 20 mol% to 45 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.6 mol%, 0.01 mol% to 0.3 mol%, 0.01 mol% to 0.2 mol%, 0.01 mol% to 0.5 mol%, 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.6 mol%, 0.05 mol% to 0.55 mol%, 0.05 mol% to 0.5 mol%, 0.05 mol% to 0.4 mol%, 0.05 mol% to 0.2 mol%, 0.05 mol% to 0.15 mol%, 0.1 mol% to 0.8 mol%, 0.1 mol% to 0.7 mol%, 0.1 mol% to 0.6 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.4 mol%, 0.15 mol% to 0.55 mol%, 0.15 mol% to 0.4 mol%, 0.2 mol% to 0.5 mol%, or 0.2 mol% to 0.3 mol% of the total lipid present in the LNPs.4 mol%, the LNPs described above have a particle size of 140 nm to 400 nm, 140 nm to 390 nm, 140 nm to 380 nm, 140 nm to 370 nm, 140 nm to 360 nm, 140 nm to 350 nm, 140 nm to 340 nm, 140 nm to 330 nm, 140 nm to 320 nm, 140 nm to 310 nm, 140 nm to 300 nm, 140 nm to 290 nm, 140 nm to 280 nm, 140 nm to 270 nm, 140 nm to 260 nm, 140 nm to 250 nm, 140 nm to 240 nm, 140 nm to 230 nm, 140 nm to 210 nm, 140 nm to 200 nm, 150 nm to 400 nm, 150 nm to 350 nm, 150 nm to 300 nm, 150 nm to 250 nm, 150 nm to 200 nm, 160 nm to 350 nm, 160 nm to 300 nm, 160 nm to 250 nm, 160 nm to 200 nm, 180 nm to 320 nm, 180 nm to 300 nm, 180 nm to 250 nm, 180 nm to 200 nm, 190 nm to 320 nm, 190 nm to 300 nm, 190 nm to 250 nm, 190 nm to 200 nm, 200 nm to 320 nm, 200 nm to 300 nm, 200 nm to 250 nm, or 200 nm to 230 nm.

[0339] In some embodiments, the LNPs comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises from 47 mol% to 55 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises from 10 mol% to 18 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises from 27 mol% to 42 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises from 0.0001 mol% to 0.8 mol%, 0.001 mol% to 0.8 mol%, 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.7 mol%, 0.01 mol% to 0.6 mol%, 0.01 mol% to 0.5 mol%, 0.01 mol% to 0.4 mol%, 0.01 mol% to 0.3 mol%, 0.01 mol% to 0.2 mol%, 0.01 mol% to 0.1 mol%, 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.7 mol%, 0.05 mol% to 0.6 mol%, 0.05 mol% to 0.5 mol%, 0.05 mol% to 0.4 mol%, 0.05 mol% to 0.3 mol%, 0.05 mol% to 0.25 mol%, 0.05 mol% to 0.2 mol%, 0.05 mol% to 0.15 mol%, 0.05 mol% to 0.1 mol%, 0.06 mol% to 0.5 mol%, 0.06 mol% to 0.4 mol%, 0.06 mol% to 0.3 mol%, 0.06 mol% to 0.2 mol%, 0.06 mol% to 0.15 mol%, 0.06 mol% to 0.1 mol%, 0.07 mol% to 0.5 mol%, 0.07 mol% to 0.4 mol%, 0.07 mol% to 0.3 mol%, 0.07 mol% to 0.2 mol%, 0.07 mol% to 0.15 mol%, 0.07 mol% to 0.1 mol%, 0.08 mol% to 0.5 mol%, 0.08 mol% to 0.4 mol%, 0.08 mol% to 0.3 mol%, 0.08 mol% to 0.2 mol%, 0.08 mol% to 0.15 mol%, 0.08 mol% to 0.1 mol%, 0.09 mol% to 0.5 mol%, 0.09 mol% to 0.4 mol%, 0.09 mol% to 0.3 mol%, 0.09 mol% to 0.2 mol%, 0.09 mol% to 0.15 mol%, 0.09 mol% to 0.1 mol%, 0.1 mol% to 0.8 mol%, 0.1 mol% to 0.7 mol%, 0.1 mol% to 0.6 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.4 mol%, 0.1 mol% to 0.3 mol%, 0.1 mol% to 0.2 mol%, 0.1 mol% to 0.15 mol%, 0.1 mol% to 0.1 mol%, 0.1 mol% to 0.05 mol%, 0.2 mol% to 0.8 mol%, 0.2 mol% to 0.7 mol%, 0.2 mol% to 0.6 mol%, 0.2 mol% to 0.5 mol%, 0.2 mol% to 0.4 mol%, 0.2 mol% to 0.3 mol%, 0.3 mol% to 0.8 mol%, 0.3 mol% to 0.7 mol%, 0.3 mol% to 0.6 mol%, 0.3 mol% to 0.5 mol%, 0.3 mol% to 0.4 mol%, 0.4 mol% to 0.8 mol%, 0.4 mol% to 0.7 mol%, 0.4 mol% to 0.6 mol%, 0.4 mol% to 0.5 mol%, 0.5 mol% to 0.8 mol%, 0.5 mol% to 0.7 mol%, 0.5 mol% to 0.6 mol%, 0.6 mol% to 0.8 mol%, 0.6 mol% to 0.7 mol%, or 0.7 mol% to 0.8 mol% of the total lipid present in the LNPs.0.1 mol% to 0.7 mol%, 0.1 mol% to 0.6 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.4 mol%, 0.1 mol% to 0.3 mol%, 0.1 mol% to 0.2 mol%, 0.1 mol% to 0.15 mol%, 0.15 mol% to 0.8 mol%, 0.15 mol% to 0.7 mol%, 0.15 mol% to 0.6 mol%, 0.15 mol% to 0.5 mol%, 0.15 mol% to 0.4 mol%, 0.15 mol% to 0.3 mol%, 0.15 mol% to 0.25 mol%, 0.2 mol% to 0.8 mol%, 0.2 mol% to 0.7 mol%, 0.2 mol% to 0.6 mol%, 0.2 mol% to 0.5 mol%, 0.2 mol% to 0.4 mol%, or 0.2 mol% to 0.35 mol%, and the LNPs have a particle size of 140 nm to 450 nm, 140 nm to 400 nm, 140 nm to 390 nm, 140 nm to 380 nm, 140 nm to 370 nm, 140 nm to 360 nm, 140 nm to 350 nm, 140 nm to 340 nm, 140 nm to 330 nm, 140 nm to 320 nm, 140 nm to 310 nm, 140 nm to 300 nm, 140 nm to 290 nm, 140 nm to 280 nm, 140 nm to 270 nm, 140 nm to 260 nm, 140 nm to 250 nm, 140 nm to 240 nm, 140 nm to 230 nm, 140 nm to 210 nm, 140 nm to 200 nm, 150 nm to 450 nm, 150 nm to 400 nm, 150 nm to 390 nm, 150 nm to 380 nm, 150 nm to 370 nm, 150 nm to 360 nm, 150 nm to 350 nm, 150 nm to 340 nm, 150 nm to 330 nm, 150 nm to 320 nm, 150 nm to 310 nm, 150 nm to 300 nm, 150 nm to 290 nm, 150 nm to 280 nm, 150 nm to 270 nm, 150 nm to 260 nm, 150 nm to 250 nm, 150 nm to 240 nm, 150 nm to 230 nm, 150 nm to 210 nm, 150 nm to 200 nm, 160 nm to 320 nm, 160 nm to 310 nm, 160 nm to 300 nm, 160 nm to 290 nm, 160 nm to 280 nm, 160 nm to 270 nm, 160 nm to 260 nm, 160 nm to 250 nm, 160 nm to 240 nm, 160 nm to 230 nm, 160 nm to 230 nm, 160 nm to 210 nm, 160 nm to 200 nm,180 nm to 320 nm, 180 nm to 310 nm, 180 nm to 300 nm, 180 nm to 290 nm, 180 nm to 280 nm, 180 nm to 270 nm, 180 nm to 260 nm, 180 nm to 250 nm, 180 nm to 240 nm, 180 nm to 230 nm, 180 nm to 230 nm, 180 nm to 210 nm, 180 nm to 200 nm, 190 nm to 320 nm, 190 nm to 310 nm, 190 nm to 300 nm, 190 nm to 290 nm, 190 nm to 280 nm, 190 nm to 270 nm, 190 nm to 260 nm, 190 nm to 250 nm, 190 nm to 240 nm, 190 nm to 230 nm, 190 nm to 230 nm, 190 nm to 210 nm, 190 nm to 200 nm, 200 nm to 320 nm, 200 nm to 310 nm, 200 nm to 300 nm, 200 nm to 290 nm, 200 nm to 280 nm, 200 nm to 270 nm, 200 nm to 260 nm, 200 nm to 250 nm, 200 nm to 240 nm, 200 nm to 230 nm, or 200 nm to 210 nm.

[0340] In some embodiments, the LNPs comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 47 mol% to 55 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 10 mol% to 18 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 27 mol% to 42 mol% of the total lipid present in the LNPs, and the polymeric lipid (e.g., a PEG lipid) comprises 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.6 mol%, 0.01 mol% to 0.3 mol%, 0.01 mol% to 0.2 mol%, 0.01 mol% to 0.5 mol%, 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.6 mol%, 0.05 mol% to 0.55 mol%, 0.05 mol% to 0.5 mol%, 0.05 mol% to 0.4 mol%, 0.05 mol% to 0.2 mol%, 0.05 mol% to 0.15 mol%, 0.1 mol% to 0.8 mol%, 0.1 mol% to 0.7 mol%, 0.1 mol% to 0.6 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.4 mol%, 0.15 mol% to 0.55 mol%, 0.15 mol% to 0.4 mol%, 0.2 mol% to 0.5 mol%, or 0.2 mol% to 0.3 mol% of the total lipid present in the LNPs.4 mol%, the particle size of the above LNPs is 140 nm-400 nm, 140 nm-390 nm, 140 nm-380 nm, 140 nm-370 nm, 140 nm-360 nm, 140 nm-350 nm, 140 nm-340 nm, 140 nm-330 nm, 140 nm-320 nm, 140 nm-310 nm, 140 nm-300 nm, 140 nm-290 nm, 140 nm-280 nm, 140 nm-270 nm, 140 nm-260 nm, 140 nm-250 nm, 140 nm-240 nm, 140 nm-230 nm, 140 nm-210 nm, 140 nm-200 nm, 150 nm-400 nm, 150 nm-350 nm, 150 nm-300 nm, 150 nm-250 nm, 150 nm-200 nm, 160 nm-350 nm, 160 nm-300 nm, 160 nm-250 nm, 160 nm-200 nm, 180 nm-320 nm, 180 nm-300 nm, 180 nm-250 nm, 180 nm-200 nm, 190 nm-320 nm, 190 nm-300 nm, 190 nm-250 nm, 190 nm-200 nm, 200 nm-320 nm, 200 nm-300 nm, 200 nm-250 nm, or 200 nm-230 nm.

[0341] The above LNPs have spleen or immune cell targeting property, which can increase the content of the active ingredient in the spleen or immune cells, increase the spleen / liver delivery ratio of the active ingredient, reduce the content of the active ingredient in the liver, and can be used for preparing a drug or a delivery carrier targeting the spleen or immune cells (e.g., a drug or a delivery carrier targeting the spleen or immune cells administered in vivo), thereby can be used for preventing or treating diseases. For example, can be used for preventing or treating diseases related to immune cells (e.g., autoimmune diseases, T cell tumors, B cell tumors, etc.), tumors or cancers, infectious diseases, etc.

[0342] The above LNPs have spleen or immune cell targeting property, which can increase the content of the active ingredient in the spleen or immune cells, increase the spleen / liver delivery ratio of the active ingredient, reduce the content of the active ingredient in the liver, and can be used for preparing a drug or a delivery carrier targeting the spleen or immune cells (e.g., a drug or a delivery carrier targeting the spleen or immune cells administered in vivo), thereby can be used for preventing or treating diseases. For example, can be used for preventing or treating diseases related to immune cells (e.g., autoimmune diseases, T cell tumors, B cell tumors, etc.), tumors or cancers, infectious diseases, etc.

[0343] The above LNPs have spleen or immune cell targeting property, which can increase the content of the active ingredient in the spleen or immune cells of the body, increase the spleen / liver delivery ratio of the active ingredient in the body, and reduce the content of the active ingredient in the liver of the body, and can be used to prepare a drug or a delivery carrier for targeting the spleen or immune cells for administration in vivo, thereby being useful for preventing or treating diseases. For example, it can be used for preventing or treating diseases related to immune cells (e.g., autoimmune diseases, T cell tumors, B cell tumors, etc.), tumors or cancers, infectious diseases, etc. The delivery effect of the above LNPs on the active substance can be determined using a method known in the art. For example, the delivery effect of the above LNPs on the active substance is determined by detecting the extent to which the above LNPs encapsulating mRNA encoding a reporter protein express the mRNA in vivo.

[0344] In addition, the present disclosure also provides another kind of lipid nanoparticles (LNPs) comprising an ionizable lipid, a helper lipid, a structural lipid, and a polymeric lipid, wherein the particle size of the LNPs is 120 nm to 1000 nm.

[0345] In some embodiments, the particle size of the above-described LNPs is as described for the previously described lipid nanoparticles. For example, the particle size of the above-described LNPs is 120 nm to 500 nm, 130 nm to 500 nm, 140 nm to 500 nm, 140 nm to 450 nm, 140 nm to 400 nm, 140 nm to 390 nm, 140 nm to 380 nm, 140 nm to 370 nm, 140 nm to 360 nm, 140 nm to 350 nm, 140 nm to 340 nm, 140 nm to 330 nm, 140 nm to 320 nm, 140 nm to 310 nm, 140 nm to 300 nm, 140 nm to 290 nm, 140 nm to 280 nm, 140 nm to 270 nm, 140 nm to 260 nm, 140 nm to 250 nm, 140 nm to 240 nm, 140 nm to 230 nm, 140 nm to 210 nm, 140 nm to 200 nm, 150 nm to 500 nm, 150 nm to 450 nm, 150 nm to 400 nm, 150 nm to 390 nm, 150 nm to 380 nm, 150 nm to 370 nm, 150 nm to 360 nm, 150 nm to 350 nm, 150 nm to 340 nm, 150 nm to 330 nm, 150 nm to 320 nm, 150 nm to 310 nm, 150 nm to 300 nm, 150 nm to 290 nm, 150 nm to 280 nm, 150 nm to 270 nm, 150 nm to 260 nm, 150 nm to 250 nm, 150 nm to 240 nm, 150 nm to 230 nm, 150 nm to 210 nm, 150 nm to 200 nm, 160 nm to 450 nm, 160 nm to 400 nm, 160 nm to 390 nm, 160 nm to 380 nm, 160 nm to 370 nm, 160 nm to 360 nm, 160 nm to 350 nm, 160 nm to 340 nm, 160 nm to 330 nm, 160 nm to 320 nm, 160 nm to 310 nm, 160 nm to 300 nm, 160 nm to 290 nm, 160 nm to 280 nm, 160 nm to 270 nm, 160 nm to 260 nm, 160 nm to 250 nm, 160 nm to 240 nm, 160 nm to 230 nm, 160 nm to 230 nm, 160 nm to 210 nm, 160 nm to 200 nm, 180 nm to 450 nm, 180 nm to 400 nm, 180 nm to 390 nm, 180 nm to 380 nm, 180 nm to 370 nm, 180 nm to 360 nm, 180 nm to 350 nm, 180 nm to 340 nm, 160 nm to 330 nm, 180 nm to 320 nm, 180 nm to 310 nm, 180 nm to 300 nm, 180 nm to 290 nm, 180 nm to 280 nm, 180 nm to 270 nm, 180 nm to 260 nm, 180 nm to 250 nm, 180 nm to 240 nm, 180 nm to 230 nm, 180 nm to 230 nm, 180 nm to 210 nm, 180 nm to 200 nm, 190 nm to 450 nm, 190 nm to 400 nm, 190 nm to 390 nm, 190 nm to 380 nm, 190 nm to 370 nm, 190 nm to 360 nm, 190 nm to 350 nm, 190 nm to 340 nm, 190 nm to 330 nm, 190 nm to 320 nm, 190 nm to 310 nm, 190 nm to 300 nm, 190 nm to 290 nm, 190 nm to 280 nm, 190 nm to 270 nm, 190 nm to 260 nm, 190 nm to 250 nm, 190 nm to 240 nm, 190 nm to 230 nm, 190 nm to 230 nm, 190 nm to 210 nm, 190 nm to 200 nm, 200 nm to 450 nm, 200 nm to 400 nm, 200 nm to 390 nm, 200 nm to 380 nm, 200 nm to 370 nm, 200 nm to 360 nm, 200 nm to 350 nm, 200 nm to 340 nm, 200 nm to 330 nm, 200 nm to 320 nm, 200 nm to 310 nm, 200180 nm to 300 nm, 180 nm to 290 nm, 180 nm to 280 nm, 180 nm to 270 nm, 180 nm to 260 nm, 180 nm to 250 nm, 180 nm to 240 nm, 180 nm to 230 nm, 180 nm to 230 nm, 180 nm to 210 nm, 180 nm to 200 nm, 180 nm to 450 nm, 180 nm to 400 nm, 180 nm to 390 nm, 180 nm to 380 nm, 180 nm to 370 nm, 180 nm to 360 nm, 180 nm to 350 nm, 180 nm to 340 nm, 160 nm to 330 nm, 190 nm to 320 nm, 190 nm to 310 nm, 190 nm to 300 nm, 190 nm to 290 nm, 190 nm to 280 nm, 190 nm to 270 nm, 190 nm to 260 nm, 190 nm to 250 nm, 190 nm to 240 nm, 190 nm to 230 nm, 190 nm to 230 nm, 190 nm to 210 nm, 190 nm to 200 nm, 200 nm to 320 nm, 200 nm to 310 nm, 200 nm to 300 nm, 200 nm to 290 nm, 200 nm to 280 nm, 200 nm to 270 nm, 200 nm to 260 nm, 200 nm to 250 nm, 200 nm to 240 nm, 200 nm to 230 nm, 200 nm to 230 nm, or 200 nm to 210 nm.

[0346] In some embodiments, the particle size of the above LNP is 130 nm to 500 nm. In some embodiments, the particle size of the above LNP is 140 nm to 500 nm. In some embodiments, the particle size of the above LNP is 140 nm to 450 nm. In some embodiments, the particle size of the above LNP is 140 nm to 400 nm. In some embodiments, the particle size of the above LNP is 140 nm to 380 nm. In some embodiments, the particle size of the above LNP is 140 nm to 350 nm. In some embodiments, the particle size of the above LNP is 140 nm to 300 nm. In some embodiments, the particle size of the above LNP is 140 nm to 280 nm. In some embodiments, the particle size of the above LNP is 140 nm to 270 nm. In some embodiments, the particle size of the above LNP is 140 nm to 260 nm. In some embodiments, the particle size of the above LNP is 140 nm to 200 nm.

[0347] In some embodiments, the particle size of the above LNP is 150 nm to 450 nm.

[0348] In some embodiments, the particle size of the above LNPs is 150 nm to 400 nm.

[0349] In some embodiments, the particle size of the above LNPs is 150 nm to 350 nm.

[0350] In some embodiments, the particle size of the above LNPs is 150 nm to 300 nm.

[0351] In some embodiments, the particle size of the above LNPs is 150 nm to 280 nm.

[0352] In some embodiments, the particle size of the above LNPs is 150 nm to 270 nm.

[0353] In some embodiments, the particle size of the above LNPs is 150 nm to 260 nm.

[0354] In some embodiments, the particle size of the above LNPs is 150 nm to 200 nm.

[0355] In some embodiments, the species, content, and ratio among the ionizable lipids, helper lipids, structural lipids, and polymer lipids of the above LNPs are as described for the lipid nanoparticles described previously.

[0356] In some embodiments, the above LNPs comprise an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymer lipid (e.g., a PEG lipid), the polymer lipid comprising 0 mol% to 0.8 mol% of the total lipid present in the LNPs, and the particle size of the LNPs is as described for the lipid nanoparticles described previously.

[0357] In some embodiments, the above LNPs comprise an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a PEG lipid (e.g., DMG-PEG), the PEG lipid comprising 0 mol% to 0.8 mol% of the total lipid present in the LNPs, and the particle size of the LNPs is as described for the lipid nanoparticles described previously.

[0358] In some embodiments, the LNPs described above comprise an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a PEG lipid (e.g., DMG-PEG), wherein the ionizable lipid comprises 30-60 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 0-30 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 10-60 mol% of the total lipid present in the LNPs, the PEG lipid comprises 0-0.8 mol% of the total lipid present in the LNPs, and the LNPs have a particle size as described above for the lipid nanoparticles.

[0359] In some embodiments, the LNPs described above comprise an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a PEG lipid (e.g., DMG-PEG), wherein the ionizable lipid comprises 30-60 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 0-30 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 10-60 mol% of the total lipid present in the LNPs, the PEG lipid comprises 0-0.8 mol% of the total lipid present in the LNPs, and the LNPs have a particle size as described above for the lipid nanoparticles.

[0360] In some embodiments, the LNPs described above comprise an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a PEG lipid (e.g., DMG-PEG), wherein the ionizable lipid comprises 30-60 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 0-30 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 10-60 mol% of the total lipid present in the LNPs, the PEG lipid comprises 0-0.8 mol% of the total lipid present in the LNPs, and the LNPs have a particle size as described above for the lipid nanoparticles.

[0361] In some embodiments, the LNPs described above comprise an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a PEG lipid (e.g., DMG-PEG), wherein the ionizable lipid comprises 30-60 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 0-30 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 10-60 mol% of the total lipid present in the LNPs, the PEG lipid comprises 0-0.8 mol% of the total lipid present in the LNPs, and the LNPs have a particle size as described above for the lipid nanoparticles.

[0362] In some embodiments, the above LNP comprises an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a PEG lipid (e.g., DMG-PEG), wherein the ionizable lipid comprises 30 mol% to 60 mol% of the total lipid present in the LNP, the helper lipid (e.g., DSPC) comprises 0 mol% to 30 mol% of the total lipid present in the LNP, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 10 mol% to 60 mol% of the total lipid present in the LNP, and the PEG lipid comprises 0.01 mol% to 0.5 mol%, 0.01 mol% to 0.2 mol%, 0.01 mol% to 0.3 mol%, 0.01 mol% to 0.15 mol%, 0.05 mol% to 0.5 mol%, 0.05 mol% to 0.2 mol%, 0.05 mol% to 0.15 mol%, 0.1 mol% to 0.7 mol%, 0.1 mol% to 0.6 mol%, or 0.2 mol% to 0.5 mol% of the total lipid present in the LNP, and the particle size of the LNP is as described for the above lipid nanoparticles.

[0363] In some embodiments, the above LNP comprises an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a PEG lipid (e.g., DMG-PEG), wherein the ionizable lipid comprises 30 mol% to 60 mol% of the total lipid present in the LNP, the helper lipid (e.g., DSPC) comprises 0 mol% to 30 mol% of the total lipid present in the LNP, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 10 mol% to 60 mol% of the total lipid present in the LNP, and the PEG lipid comprises 0.01 mol% to 0.5 mol%, 0.01 mol% to 0.2 mol%, 0.01 mol% to 0.3 mol%, 0.01 mol% to 0.15 mol%, 0.05 mol% to 0.5 mol%, 0.05 mol% to 0.2 mol%, 0.05 mol% to 0.15 mol%, 0.1 mol% to 0.7 mol%, 0.1 mol% to 0.6 mol%, or 0.2 mol% to 0.5 mol% of the total lipid present in the LNP, and the particle size of the LNP is as described for the above lipid nanoparticles.

[0364] In some embodiments, the above LNP comprises an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a PEG lipid (e.g., DMG-PEG), wherein the ionizable lipid comprises 45 mol% to 55 mol% of the total lipid present in the LNP, the helper lipid (e.g., DSPC) comprises 8 mol% to 25 mol% of the total lipid present in the LNP, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 20 mol% to 45 mol% of the total lipid present in the LNP, and the PEG lipid comprises 0.01 mol% to 0.5 mol%, 0.01 mol% to 0.2 mol%, 0.01 mol% to 0.3 mol%, 0.01 mol% to 0.15 mol%, 0.05 mol% to 0.5 mol%, 0.05 mol% to 0.2 mol%, 0.05 mol% to 0.15 mol%, 0.1 mol% to 0.7 mol%, 0.1 mol% to 0.6 mol%, or 0.2 mol% to 0.5 mol% of the total lipid present in the LNP, and the particle size of the LNP is as described for the above lipid nanoparticles.

[0365] In some embodiments, the above LNP comprises an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a PEG lipid (e.g., DMG-PEG), wherein the ionizable lipid comprises 45 mol% to 55 mol% of the total lipid present in the LNP, the helper lipid (e.g., DSPC) comprises 8 mol% to 25 mol% of the total lipid present in the LNP, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 20 mol% to 45 mol% of the total lipid present in the LNP, and the PEG lipid comprises 0.01 mol% to 0.5 mol%, 0.01 mol% to 0.2 mol%, 0.01 mol% to 0.3 mol%, 0.01 mol% to 0.15 mol%, 0.05 mol% to 0.5 mol%, 0.05 mol% to 0.2 mol%, 0.05 mol% to 0.15 mol%, 0.1 mol% to 0.7 mol%, 0.1 mol% to 0.6 mol%, or 0.2 mol% to 0.5 mol% of the total lipid present in the LNP, and the particle size of the LNP is as described for the above lipid nanoparticles.

[0366] In some embodiments, the LNPs described above comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a polymeric lipid (e.g., a PEG lipid), wherein the ionizable lipid comprises 47 mol% to 55 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 10 mol% to 18 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 27 mol% to 42 mol% of the total lipid present in the LNPs, the polymeric lipid (e.g., a PEG lipid) comprises 0.01 mol% to 0.5 mol%, 0.01 mol% to 0.2 mol%, 0.01 mol% to 0.3 mol%, 0.01 mol% to 0.15 mol%, 0.05 mol% to 0.5 mol%, 0.05 mol% to 0.2 mol%, 0.05 mol% to 0.15 mol%, 0.1 mol% to 0.7 mol%, 0.1 mol% to 0.6 mol%, or 0.2 mol% to 0.5 mol% of the total lipid present in the LNPs, and the LNPs have a particle size as described for the lipid nanoparticles above.

[0367] In some embodiments, the LNPs described above comprise: an ionizable lipid, a helper lipid (e.g., DSPC), a structural lipid (e.g., a non-plant sterol (e.g., cholesterol)), and a PEG lipid, wherein the ionizable lipid comprises 47 mol% to 55 mol% of the total lipid present in the LNPs, the helper lipid (e.g., DSPC) comprises 10 mol% to 18 mol% of the total lipid present in the LNPs, the structural lipid (e.g., a non-plant sterol (e.g., cholesterol)) comprises 27 mol% to 42 mol% of the total lipid present in the LNPs, the PEG lipid comprises 0.01 mol% to 0.5 mol%, 0.01 mol% to 0.2 mol%, 0.01 mol% to 0.3 mol%, 0.01 mol% to 0.15 mol%, 0.05 mol% to 0.5 mol%, 0.05 mol% to 0.2 mol%, 0.05 mol% to 0.15 mol%, 0.1 mol% to 0.7 mol%, 0.1 mol% to 0.6 mol%, or 0.2 mol% to 0.5 mol% of the total lipid present in the LNPs, and the LNPs have a particle size as described for the lipid nanoparticles above.

[0368] In some embodiments, the LNPs of any of the above embodiments further comprise an active ingredient. The active ingredient is associated with and / or encapsulated in the LNPs. The active ingredient encapsulated in and / or associated with the LNPs is delivered with the LNPs into the spleen or immune cells.

[0369] In some embodiments, the active ingredient contained in the above-mentioned LNPs is an active ingredient that needs to be delivered to the spleen.

[0370] In some embodiments, the above-mentioned LNPs encapsulate an active ingredient that needs to be delivered to the spleen.

[0371] Due to the spleen targeting property of the above-mentioned LNPs, after the above-mentioned LNPs enter the body, the active ingredient is delivered to the spleen due to being carried by the LNPs. The spleen is the largest lymphoid organ, and there are a large number of immune cells. The active ingredient delivered to the spleen can be taken up by immune cells and delivered to immune cells. Therefore, in some embodiments, the active ingredient is delivered to immune cells. In some embodiments, the active ingredient is delivered to one or more immune cells selected from the group consisting of B cells, T cells, dendritic cells, macrophages, monocytes, NK cells (e.g., immature NK cells, activated NK cells), and NKT cells. In some embodiments, the B cells to which the active ingredient is delivered are selected from one or more of the group consisting of plasmablasts, plasma cells, lymphoplasmacytoid cells, memory B cells, B-2 cells, and regulatory B cells. In some embodiments, the T cells to which the active ingredient is delivered are selected from one or more of the group consisting of killer T cells, helper T cells, regulatory T cells, naive T cells, effector T cells, and memory T cells. In some embodiments, the immune cells to which the active ingredient is delivered are primate immune cells. In one alternative specific example, the immune cells to which the active ingredient is delivered are human immune cells.

[0372] The specific functions and types of the active ingredients contained in the above-mentioned LNPs can be determined according to the application scenario and its principle (e.g., the efficacy of a specific drug and the principle of achieving the efficacy). They can be substances that directly exert an effect after being delivered to the spleen or immune cells by the LNPs (e.g., antisense oligonucleotides, tRNA, rRNA, snRNAs, gRNA, siRNA, microRNA, microRNA binding sites, non-nucleic acid small molecule drugs, chemotherapeutic drugs, polypeptides or proteins, etc.), or substances that indirectly exert an effect after being delivered to the spleen or immune cells by the LNPs by first synthesizing the actual substance that exerts an effect using the cellular environment (e.g., nucleic acids (e.g., mRNA), dsRNA, shRNA, etc. that encode a polypeptide or protein of interest).

[0373] In some embodiments, the active ingredient contained in the above-mentioned LNPs comprises or is one or more of the following: nucleic acids, non-nucleic acid small molecules, chemotherapeutic drugs, polypeptides, and proteins.

[0374] In some embodiments, the active ingredient contained in the LNPs described above comprises or is a nucleic acid. By encapsulating or binding to the LNPs, the nucleic acid is less likely to be degraded before reaching the spleen after entering the body, thus enabling it to function in the spleen. It can be appreciated that the type of nucleic acid contained in the active ingredient contained in the LNPs described above is not particularly limited. In some embodiments, the nucleic acid contained in the active ingredient is RNA. For example, mRNA, microRNA, microRNA binding site, tRNA, dsRNA, siRNA, snRNA, shRNA, sgRNA, antisense oligonucleotide, rRNA, etc. In some embodiments, the nucleic acid contained in the active ingredient is DNA. For example, ssDNA, expression vector, etc. In some embodiments, the nucleic acid contained in the active ingredient is a hybrid chain of DNA-RNA. In addition, the nucleic acid contained in the active ingredient contained in the LNPs described above can all be RNA (for example, a mixture of sgRNA and Cas9 mRNA), all be DNA, all be a hybrid chain of DNA-RNA, or be a mixture of at least two of DNA, RNA, and a hybrid chain of DNA-RNA (for example, the active ingredient is a mixture of sgRNA and a plasmid expressing Cas9).

[0375] In some embodiments, the active ingredient contained in the LNPs described above is a nucleic acid, a non-nucleic acid small molecule, a chemotherapeutic drug, a polypeptide, or a protein.

[0376] In some embodiments, the active ingredient contained in the LNPs described above is a mixture of a nucleic acid and a non-nucleic acid small molecule compound, or a conjugate of a nucleic acid and a non-nucleic acid small molecule compound. In some embodiments, the active ingredient contained in the LNPs described above is a mixture of a nucleic acid and a polypeptide or a protein, for example, sgRNA and Cas9. In some embodiments, the active ingredient contained in the LNPs described above is a mixture of a non-nucleic acid small molecule compound and a polypeptide or a protein. In some embodiments, the active ingredient contained in the LNPs described above is a conjugate of a non-nucleic acid small molecule compound and a polypeptide or a protein (for example, an antibody drug conjugate).

[0377] In some embodiments, the active ingredient contained in the LNPs described above is used to increase the amount of a protein of interest. By delivering the active ingredient (for example, a polypeptide or protein of interest, mRNA encoding a polypeptide or protein of interest, a vector (for example, a plasmid) expressing a polypeptide or protein of interest) through the LNPs described above with spleen targeting and immune cell targeting, the amount of the active ingredient delivered to the spleen or immune cells (for example, the content, activity, or expression amount of the active ingredient in the spleen) can be increased, thus the amount of the protein of interest in the immune cells, spleen, or other tissues or organs can be increased, which in turn facilitates the prevention or treatment of diseases.

[0378] A polypeptide or protein of interest refers to a therapeutically or pharmaceutically active polypeptide or protein having a prophylactic or therapeutic effect that is desired or beneficial in the spleen.

[0379] In some application scenarios where immune responses are desired to be activated or enhanced (e.g., cancer treatment, prevention or treatment of infectious diseases), the active ingredient contained in the above LNPs is used to activate immune cells or enhance immune cell activity. The amount of the active ingredient in the immune cells is increased by delivering the active ingredient through the above LNPs, thereby increasing the amount of the protein of interest, which in turn is more conducive to activating immune cells or enhancing immune cell activity. Therefore, in some embodiments, the active ingredient contained in the above LNPs for increasing the amount of the protein of interest comprises a substance that activates immune cells or enhances immune cell activity. For example, one or more of the following polypeptides or proteins, or a nucleic acid (e.g., mRNA or expression vector) encoding one or more of the following polypeptides or proteins: an antigen, a cytokine, a chemokine or chemokine receptor, a costimulatory factor, a T cell receptor (TcR), a chimeric antigen receptor (CAR), a recruiting factor, a transcription factor, an effector molecule, an MHC molecule, and an enzyme.

[0380] In some embodiments, the active ingredient of the above LNPs comprises or is a cytokine that stimulates immune cell activation or enhances immune cell activity or a nucleic acid (e.g., mRNA or expression vector) encoding the cytokine. In some embodiments, the cytokine that stimulates immune cell activation or enhances immune cell activity comprises one or more of the following: IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-12, IL-13, IL-15, IL-17, IL-18, IL-21, IL-23, TNF-a, TNF-b, IFN-a, and IFN-b.

[0381] In some embodiments, the active ingredient of the above LNPs comprises or is a cytokine that promotes T cell activation or differentiation or a nucleic acid (e.g., mRNA or expression vector) encoding the cytokine that promotes T cell activation or differentiation. In some embodiments, the cytokine that promotes T cell activation or differentiation comprises one or more of the following: IL-2, IL-4, IL-12, IL-13, IL-15, and IFN-a. In some embodiments, the active ingredient contained in the above LNPs comprises a cytokine that promotes a Th2 response or a nucleic acid (e.g., mRNA or expression vector) encoding the cytokine. In some embodiments, the cytokine that promotes a Th2 response comprises one or both of the following: IL-4 and IL-10.

[0382] In some embodiments, the active ingredient contained by the above LNPs comprises or is a cytokine that promotes B cell activation or a nucleic acid (e.g., mRNA or expression vector) encoding a cytokine that promotes B cell activation. In some embodiments, the cytokine that promotes B cell activation comprises one or more of the following: IL-4, IL-5, IL-6, IL-10, IL-13, and IFN.

[0383] In some embodiments, the active ingredient contained by the above LNPs comprises or is a pro-inflammatory cytokine or a nucleic acid (e.g., mRNA or expression vector) encoding a pro-inflammatory cytokine. Pro-inflammatory cytokines can be used in treatments that require stimulation of inflammatory responses (e.g., pro-inflammatory cytokines can increase anti-tumor immunity when used in cancer treatment). In some embodiments, the pro-inflammatory cytokine comprises one or more of the following: IL-1, IL-6, IL-17, IL-18, IL-23, TNF-a, IFN-a, IFN-b, and interferon-gamma.

[0384] In some embodiments, the active ingredient contained by the above LNPs comprises or is a chemokine or a chemokine receptor, or a nucleic acid (e.g., mRNA or expression vector) encoding a chemokine or a chemokine receptor. Chemokines are involved in regulating inflammatory and immune responses, and can control the movement of inflammatory cells (including granulocytes, monocytes) as well as modulate a variety of immune cells (including lymphocytes, natural killer cells, and dendritic cells). In addition, chemokines have an impact on the proliferation and invasive properties of cancer cells.

[0385] In some embodiments, the active ingredient contained by the above LNPs comprises or is a chemokine or a chemokine receptor that acts on regulatory T cells, or a nucleic acid (e.g., mRNA or expression vector) encoding the chemokine or the chemokine receptor. In some embodiments, the chemokine or the chemokine receptor that acts on regulatory T cells comprises one or more of the following: CCL22, CCL28, CCR4, and CCR10.

[0386] In some embodiments, the active ingredient contained by the above LNPs comprises or is a chemokine or a chemokine receptor that acts on cytotoxic T cells, or a nucleic acid (e.g., mRNA or expression vector) encoding the chemokine or the chemokine receptor. In some embodiments, the chemokine or the chemokine receptor that acts on cytotoxic T cells comprises one or more of the following: CXCL9, CXCL10, CXCL11, and CXCR3.

[0387] In some embodiments, the active ingredient contained in the above LNP comprises or is a chemokine or chemokine receptor that acts on natural killer cells, or a nucleic acid (e.g., mRNA or expression vector) encoding the chemokine or chemokine receptor. In some embodiments, the chemokine or chemokine receptor that acts on natural killer cells comprises one or more of: CXCL9, CXCL10, CXCL11, CCL3, CCL4, CCL5, CCL2, CCL8, CCL12, CCL13, CCL19, CCL21, CX3CL1, CXCR3, CCR1, CCR5, CCR2, and CX3CR1.

[0388] In some embodiments, the active ingredient contained in the above LNP comprises or is a chemokine or chemokine receptor that acts on immature dendritic cells, or a nucleic acid (e.g., mRNA or expression vector) encoding the chemokine or chemokine receptor. In some embodiments, the chemokine or chemokine receptor that acts on immature dendritic cells comprises one or more of: CCL3, CCL4, CCL5, CCL2, CCL7, CCL8, CCL22, CCL1, CCL17, CXCL12, CCR1, CCR2, CCR4, CCR5, CCR6, CCR8, and CXCR4.

[0389] In some embodiments, the active ingredient contained in the above LNP comprises or is a chemokine or chemokine receptor that acts on mature dendritic cells, or a nucleic acid (e.g., mRNA or expression vector) encoding the chemokine or chemokine receptor. In some embodiments, the chemokine or chemokine receptor that acts on mature dendritic cells comprises one or more of: CCL19, CCL21, CXCL12, CCR7, and CXCR4.

[0390] In some embodiments, the active ingredient contained in the above LNP comprises or is a chemokine or chemokine receptor that acts on tumor-associated macrophages, or a nucleic acid (e.g., mRNA or expression vector) encoding the chemokine or chemokine receptor. In some embodiments, the chemokine or chemokine receptor that acts on tumor-associated macrophages comprises one or more of: CCL2, CCL7, CCL8, CCL3, CCL4, CCL5, CXCL12, CCR2, CCR5, and CXCR4.

[0391] In some embodiments, the active ingredient contained in the above-mentioned LNPs comprises or is a recruitment factor that stimulates immune cell activation or enhances immune cell activity or a nucleic acid (e.g., mRNA or expression vector) encoding the recruitment factor. As used herein, “recruitment factor” refers to any protein that facilitates the recruitment of immune cells to a desired location (e.g., to a tumor site or an inflammation site). In some embodiments, the recruitment factor that stimulates immune cell activation or enhances immune cell activity comprises one or more of the following: CXCR3, CXCR5, CCR5, CCL5, CXCL10, CXCL12, CXCL16, and IFN-β.

[0392] In some embodiments, the active ingredient contained in the above-mentioned LNPs for activating immune cells or enhancing immune cell activity comprises or is a pathogenic antigen, an RNA (e.g., mRNA) encoding the pathogenic antigen, or a DNA (e.g., expression vector) expressing the pathogenic antigen. When the active ingredient (pathogenic antigen, RNA encoding the pathogenic antigen, or expression vector expressing the pathogenic antigen) is delivered using the above-mentioned LNPs, the amount of the pathogenic antigen, RNA encoding the pathogenic antigen, or expression vector expressing the pathogenic antigen delivered to the spleen is increased, thereby increasing the amount of the pathogenic antigen in the spleen, which in turn increases the amount of the protein of interest (e.g., cytokine, specific antibody, etc.) by increasing the contact of immune cells (e.g., T cells, B cells, dendritic cells, etc.) in the spleen with the pathogenic antigen.

[0393] In some embodiments, the pathogenic antigen is derived from a pathogenic organism that is capable of causing an immune response in a subject (e.g., a mammalian subject, further for example, a human). In some embodiments, the pathogenic organism comprises or is one or more of the following: a bacterium, a virus, a fungus, and a protozoan (e.g., a single-celled organism, a multi-cellular organism). In some embodiments, the pathogenic antigen comprises or is a surface antigen, an immunogenic fragment, a variant, or a variant of an immunogenic fragment, e.g., a protein located on the surface of a virus, a bacterium, or a protozoan, an immunogenic fragment (e.g., an external portion of a surface antigen) thereof, a variant, or a variant of an immunogenic fragment. In some embodiments, the pathogenic antigen comprises or is a polypeptide or protein derived from a pathogen associated with an infectious disease. In some embodiments, the pathogenic antigen is selected from the group consisting of the antigens derived from a pathogen described in WO2018 / 078053A1 at pages 21-35, the antigens derived from a pathogen described in WO2019 / 077001A1 at page 57, paragraph 3 - page 63, paragraph 2, the antigens derived from a pathogen described in WO2013 / 120628A1 at page 32, line 26 - page 34, line 27, and the antigens described in WO2013 / 120628A1 at page 34, line 29 - page 59, line 5.

[0394] In some embodiments, the active ingredient contained in the LNPs described above for activating immune cells or enhancing immune cell activity comprises an antigen receptor (e.g., a chimeric antigen receptor or a T cell antigen receptor) or a nucleic acid (e.g., mRNA or an expression vector) encoding an antigen receptor. The use of the LNPs described above to deliver an antigen receptor or a nucleic acid encoding an antigen receptor can increase the amount of the antigen receptor or the nucleic acid encoding the antigen receptor delivered to the spleen, which can increase the amount of antigen receptor-immune cells in the spleen, and in turn, increase the specificity and killing power of the drug against tumor cells.

[0395] In some embodiments, the active ingredient contained in the LNPs described above comprises or is: a chimeric antigen receptor or a T cell antigen receptor, an RNA (e.g., mRNA) encoding a chimeric antigen receptor or a T cell antigen receptor, or a DNA (e.g., an expression vector) expressing a chimeric antigen receptor or a T cell antigen receptor.

[0396] A chimeric antigen receptor is an artificial transmembrane receptor that enables an immune effector cell (e.g., a T cell, a natural killer cell, or other immune cell) comprising the chimeric antigen receptor to have specificity for a ligand of the chimeric antigen receptor, and upon specific recognition and binding of the ligand by the chimeric antigen receptor, the effector cell is activated. A chimeric antigen receptor generally comprises a tumor antigen binding region (e.g., a scFv), an extracellular hinge region, a transmembrane region, and an intracellular immunoreceptor tyrosine-based activation motif (e.g., CD3 zeta, CD28, CD137 (also known as 4-lBB), CD134 (also known as OX40), and CD278 (also known as ICOS), etc.). A tumor antigen is generally only present on the surface of tumor cells and not expressed in normal tissues, and thus the chimeric antigen receptor can serve to direct the CAR-T cell to only have killing effects on tumor cells.

[0397] A T cell receptor (TcR) used in therapy is generally a specific receptor on the surface of a T cell that is capable of recognizing and binding a tumor antigen, and the TcR induces activation of the T cell and exerts an adaptive immune effector function by specifically recognizing and binding a tumor antigen presented by a major histocompatibility complex (MHC) on the surface of an antigen presenting cell (APC) to form a TcR-antigen-MHC complex.

[0398] In some embodiments, the chimeric antigen receptor is capable of recognizing one or more of the following tumor antigens: CD19, CD20, CD22, CD30, CD33, CD123, CD138, CD171, CEA, EGFR, HER2, mesothelin, and PSMA.

[0399] In some embodiments, the TcR is capable of recognizing one or more of the following tumor antigens: CD19, CD20, CD22, CD30, CD33, CD123, CD138, CD171, CEA, EGFR, HER2, mesothelin, and PSMA.

[0400] In some embodiments, the tumor antigen is selected from the group consisting of the tumor antigens described in WO2018 / 078053A1, pages 47-51, but is not limited thereto.

[0401] In some embodiments, the LNPs described above comprise an active ingredient for reducing the amount of a protein of interest. The LNPs described above have spleen targeting property, and delivery of the active ingredient (e.g., gene knockout, gene silencing, or antibody, etc.) using the LNPs described above can increase the amount of the active ingredient delivered to the spleen, thereby increasing the effect of the active ingredient, and in turn, can reduce (i.e., decrease, inhibit, down-regulate) the expression of the protein of interest in the spleen or other tissues or organs.

[0402] In some application scenarios of inhibiting immune cell activation or immune cell activity (e.g., autoimmune diseases, allergies, and transplantation), the LNPs described above comprise an active ingredient for inhibiting immune cell activation or immune cell activity. Delivery of the active ingredient using the LNPs described above can increase the amount of the active ingredient in the immune cells, and in turn, can increase the effect of inhibiting immune cell activation or degrading immune cell activity. For example, for siRNA drugs, the increase of the active ingredient delivered to the spleen can increase the effect of the active molecule on the target mRNA (interfering with the expression of the target mRNA), thereby reducing the production of the protein (e.g., a cytokine that activates immune cells) corresponding to the target mRNA, and in turn, inhibiting immune cell activation or reducing immune cell activity, and reducing the production of the protein of interest (e.g., a cytokine) by immune cells (e.g., T cells, B cells, dendritic cells). For example, for drugs for autoimmune diseases, the increase of the active ingredient (e.g., a nucleic acid (e.g., a 1-methylpseudouridine-modified mRNA) encoding a self-antigen) delivered to the spleen can increase the antigen-specific immune tolerance of the body, and in turn, can selectively weaken the damage to the body caused by autoimmunity.

[0403] In some embodiments, the active ingredient for reducing the amount of a protein of interest comprised in the LNPs described above is an active ingredient for inhibiting an immune response. In some embodiments, the active ingredient for reducing the amount of a protein of interest comprised in the LNPs described above is an active ingredient for inhibiting an immune response of lymphocytes. In some embodiments, the active ingredient for reducing the amount of a protein of interest comprised in the LNPs described above inhibits an immune response by one or more of the following: B cell activity modulation, B cell depletion, antigen tolerance, inhibition of antibody production, B regModulation of differentiation, T effector cell modulation, induction of effector T cell anergy (e.g., induction of effector T cell migration, suppression of cytokine or cytokine receptor expression), and effector T cell exhaustion.

[0404] In some embodiments, the protein of interest inhibited by the active ingredient of the above-described LNPs is selected from one or more of the following...

Claims

1. Use of a lipid nanoparticle (LNPs) comprising an active ingredient, an ionizable lipid, a helper lipid, a structural lipid, and a polymer lipid in an amount of 0 mol% to 0.8 mol% of the total lipid present in the LNPs, for the manufacture of a medicament targeting the spleen or immune cells.

2. Use of a lipid nanoparticle (LNPs) comprising an active ingredient, an ionizable lipid, a helper lipid, a structural lipid, and a polymer lipid, wherein the LNPs have a particle size of 130 nm to 500 nm, for the manufacture of a medicament targeting the spleen or immune cells.

3. Use of the LNPs according to claim 1, wherein the LNPs have a particle size of 130 nm to 500 nm, for the manufacture of a medicament targeting the spleen or immune cells.

4. Use of the LNPs according to claim 2, wherein the polymer lipid is in an amount of 0 mol% to 0.8 mol% of the total lipid present in the LNPs, for the manufacture of a medicament targeting the spleen or immune cells.

5. Use of the LNPs according to any one of claims 1 to 4, for the manufacture of a medicament targeting the spleen or immune cells, wherein the LNPs are capable of increasing the content of the active ingredient in the spleen, increasing the content of the active ingredient in immune cells, increasing the spleen / liver delivery ratio of the active ingredient, and / or decreasing the content of the active ingredient in the liver, as compared to other LNPs having a polymer lipid content greater than the maximum of the range of the polymer lipid content of the LNPs.

6. Use of the LNPs of any one of claims 1 and 3-5 for the manufacture of a medicament for targeting the spleen or immune cells, the polymeric lipid comprising 0.01 mol% to 0.8 mol%, 0.01 mol% to 0.7 mol%, 0.01 mol% to 0.6 mol%, 0.01 mol% to 0.5 mol%, 0.01 mol% to 0.4 mol%, 0.01 mol% to 0.3 mol%, 0.01 mol% to 0.2 mol%, 0.01 mol% to 0.1 mol%, 0.05 mol% to 0.8 mol%, 0.05 mol% to 0.7 mol%, 0.05 mol% to 0.6 mol%, 0.05 mol% to 0.5 mol%, 0.05 mol% to 0.4 mol%, 0.05 mol% to 0.3 mol%, 0.05 mol% to 0.25 mol%, 0.05 mol% to 0.2 mol%, 0.05 mol% to 0.15 mol%, 0.05 mol% to 0.1 mol%, 0.1 mol% to 0.8 mol%, 0.1 mol% to 0.7 mol%, 0.1 mol% to 0.6 mol%, 0.1 mol% to 0.5 mol%, 0.1 mol% to 0.4 mol%, 0.1 mol% to 0.3 mol%, 0.1 mol% to 0.2 mol%, 0.1 mol% to 0.15 mol%, 0.15 mol% to 0.8 mol%, 0.15 mol% to 0.7 mol%, 0.15 mol% to 0.6 mol%, 0.15 mol% to 0.5 mol%, 0.15 mol% to 0.4 mol%, 0.15 mol% to 0.3 mol%, 0.15 mol% to 0.25 mol%, 0.2 mol% to 0.8 mol%, 0.2 mol% to 0.7 mol%, 0.2 mol% to 0.6 mol%, 0.2 mol% to 0.5 mol%, 0.2 mol% to 0.4 mol%, or 0.2 mol% to 0.35 mol% of the total lipid present in the LNPs; Preferably, the polymeric lipid comprises 0.01 mol% to 0.5 mol% of the total lipid present in the LNPs; Preferably, the polymeric lipid comprises 0.01 mol% to 0.2 mol% of the total lipid present in the LNPs; Preferably, the polymeric lipid comprises 0.05 mol% to 0.2 mol% of the total lipid present in the LNPs; Preferably, the polymeric lipid comprises 0.06 mol% to 0.3 mol% of the total lipid present in the LNPs; Preferably, the polymeric lipid comprises 0.06 mol% to 0.2 mol% of the total lipid present in the LNPs; Preferably, the polymeric lipid comprises 0.06 mol% to 0.15 mol% of the total lipid present in the LNPs; Preferably, the polymeric lipid comprises 0.07 mol% to 0.3 mol% of the total lipid present in the LNPs; Preferably, the polymeric lipid comprises 0.07 mol% to 0.2 mol% of the total lipid present in the LNPs; Preferably, the polymeric lipid comprises 0.07 mol% to 0.15 mol% of the total lipid present in the LNPs; Preferably, the polymeric lipid comprises 0.08 mol% to 0.3 mol% of the total lipid present in the LNPs; Preferably, the polymeric lipid comprises 0.08 mol% to 0.2 mol% of the total lipid present in the LNPs; Preferably, the polymeric lipid comprises 0.08 mol% to 0.15 mol% of the total lipid present in the LNPs.

7. Use of the LNPs according to any one of claims 2-5 having a particle size of 140 nm to 500 nm, 140 nm to 450 nm, 140 nm to 400 nm, 140 nm to 390 nm, 140 nm to 380 nm, 140 nm to 370 nm, 140 nm to 360 nm, 140 nm to 350 nm, 140 nm to 340 nm, 140 nm to 330 nm, 140 nm to 320 nm, 140 nm to 310 nm, 140 nm to 300 nm, 140 nm to 290 nm, 140 nm to 280 nm, 140 nm to 270 nm, 140 nm to 260 nm, 140 nm to 250 nm, 140 nm to 240 nm, 140 nm to 230 nm, 140 nm to 210 nm, 140 nm to 200 nm, 150 nm to 450 nm, 150 nm to 400 nm, 150 nm to 390 nm, 150 nm to 380 nm, 150 nm to 370 nm, 150 nm to 360 nm, 150 nm to 350 nm, 150 nm to 340 nm, 150 nm to 330 nm, 150 nm to 320 nm, 150 nm to 310 nm, 150 nm to 300 nm, 150 nm to 290 nm, 150 nm to 280 nm, 150 nm to 270 nm, 150 nm to 260 nm, 150 nm to 250 nm, 150 nm to 240 nm, 150 nm to 230 nm, 150 nm to 210 nm, 150 nm to 200 nm, 160 nm to 320 nm, 160 nm to 310 nm, 160 nm to 300 nm, 160 nm to 290 nm, 160 nm to 280 nm, 160 nm to 270 nm, 160 nm to 260 nm, 160 nm to 250 nm, 160 nm to 240 nm, 160 nm to 230 nm, 160 nm to 230 nm, 160 nm to 210 nm, 160 nm to 200 nm, 180 nm to 320 nm, 180 nm to 310 nm, 180 nm to 300 nm, 180 nm to 290 nm, 180 nm to 280 nm, 180 nm to 270 nm, 180 nm to 260 nm, 180 nm to 250 nm, 180 nm to 240 nm, 180 nm to 230 nm, 180 nm to 230 nm, 180 nm to 210 nm, 180 nm to 200 nm, 190 nm to 320 nm, 190 nm to 310 nm, 190 nm to 300 nm, 190 nm to 290 nm, 190 nm to 280 nm, 190 nm to 270 nm, 190 nm to 260 nm, 190 nm to 250 nm, 190 nm to 240 nm, 190 nm to 230 nm, 190 nm to 230 nm, 190 nm to 210 nm, 190 nm to 200 nm, 200 nm to 320 nm, 200 nm to 310 nm, 200 nm to 300 nm, 200 nm to 290 nm, 200 nm to 280 nm, 200 nm to 270 nm, 200 nm to 260 nm, 200 nm to 250 nm, 200 nm to 240 nm, 200 nm to 230 nm, 200 nm to 230 nm, 200 nm to 210 nm, 200 nm to 210 nm, 210 nm to 320 nm, 210 nm to 310 nm, 210 nm to 300 nm, 210 nm to 290 nm, 210 nm to 280 nm, 210 nm to 270 nm, 210 nm to 260 nm, 210 nm to 250 nm, 210 nm to 240 nm, 210 nm to 230 nm, 210 nm to 230 nm, 220 nm to 320 nm, 220 nm to 310 nm, 220 nm to 300 nm, 220 nm to 290 nm, 220 nm to 280 nm, 220 nm to 270 nm, 220 nm to 260 nm, 220 nm to 250 nm, 220 nm to 240 nm, 220 nm to 230 nm, 220 nm to 230 nm, 230 nm to 320 nm, 230 nm to 310 nm, 230 nm to 300 nm, 230 nm to 290 nm, 230 nm to 280 nm, 230 nm to 270 nm, 230 nm to 260 nm, 230 nm to 250 nm, 230 nm to 240 nm, 240 nm to 320 nm, 240 nm to 310 nm, 240 nm to 300 nm, 240 nm to 290 nm, 240 nm to 280 nm, 240 nm to 270 nm, 240 nm to 260 nm, 240 nm to 250 nm, 250 nm to 320 nm, 250 nm to 310 nm, 250 nm to 300 nm, 250 nm to 290 nm, 250 nm to 280 nm, 250 nm to 270 nm, 250 nm to 260 nm, 260 nm to 320 nm, 260 nm to 310 nm, 260 nm to 300 nm, 260 nm to 290 nm, 260 nm to 280 nm, 260 nm to 270 nm, 270 nm to 320 nm, 270 nm to 310 nm, 270 nm to 300 nm, 270 nm to 290 nm, 270 nm to 280 nm, 280 nm to 320 nm, 280 nm to 310 nm, 280 nm to 300 nm, 280 nm to 290 nm, 290 nm to 320 nm, 290 nm to 310 nm, 290 nm to 300 nm, 300 nm to 320 nm, 300 nm to 310 nm, 310 nm to 320 nm, or 320 nm for the preparation of a medicament for targeting the spleen or immune cells.190 nm to 230 nm, 190 nm to 210 nm, 190 nm to 200 nm, 200 nm to 320 nm, 200 nm to 310 nm, 200 nm to 300 nm, 200 nm to 290 nm, 200 nm to 280 nm, 200 nm to 270 nm, 200 nm to 260 nm, 200 nm to 250 nm, 200 nm to 240 nm, 200 nm to 230 nm, or 200 nm to 210 nm; Preferably, the LNPs have a particle size of 140 nm to 400 nm; Preferably, the LNPs have a particle size of 150 nm to 300 nm; Preferably, the LNPs have a particle size of 150 nm to 280 nm; Preferably, the LNPs have a particle size of 150 nm to 200 nm; Preferably, the LNPs have a particle size of 200 nm to 400 nm; Preferably, the LNPs have a particle size of 200 nm to 340 nm; Preferably, the LNPs have a particle size of 250 nm to 340 nm.

8. Use of the LNPs according to any one of claims 1 to 7 for the manufacture of a medicament for targeting the spleen or immune cells, the ionizable lipid comprising 30 mol% to 60 mol% of the total lipid present in the LNPs, the helper lipid comprising 0 mol% to 30 mol% of the total lipid present in the LNPs, the structural lipid comprising 10 mol% to 60 mol% of the total lipid present in the LNPs; Preferably, the ionizable lipid comprises 45 mol% to 55 mol% of the total lipid present in the LNPs, the helper lipid comprises 8 mol% to 25 mol% of the total lipid present in the LNPs, the structural lipid comprises 20 mol% to 45 mol% of the total lipid present in the LNPs; Preferably, the ionizable lipid comprises 47 mol% to 55 mol% of the total lipid present in the LNPs, the helper lipid comprises 10 mol% to 28 mol% of the total lipid present in the LNPs, the structural lipid comprises 27 mol% to 42 mol% of the total lipid present in the LNPs.

9. Use of the LNPs according to any one of claims 1 to 8 for the manufacture of a medicament for targeting the spleen or immune cells, the polymeric lipid being a PEG lipid; Preferably, the PEG lipid is selected from one or more of DMG-PEG, DSG-PEG and DPG-PEG.

10. Use of the LNPs according to any one of claims 1 to 9 for the manufacture of a medicament for targeting the spleen or immune cells, the helper lipid being a phospholipid; Preferably, the helper lipid is selected from one or more of DSPC, DOPE, DOPC and DOPS; Preferably, the helper lipid is DSPC.

11. Use of the LNPs according to any one of claims 1 to 10 for the manufacture of a medicament for targeting the spleen or immune cells, the structural lipid being cholesterol.

12. Use of the LNPs according to any one of claims 1 to 11 for the manufacture of a medicament for targeting the spleen or immune cells, the immune cells comprising one or more of the following: B cells, T cells, NK cells, DC cells, neutrophils, eosinophils, basophils, mast cells, monocytes and macrophages; Preferably, the immune cells are B cells, T cells or NK cells.

13. Use of the LNPs according to any one of claims 1 to 12 for the manufacture of a medicament for targeting the spleen or immune cells, the active ingredient being a nucleic acid, a protein, a polypeptide or a small molecule.

14. Use of the LNPs according to any one of claims 1 to 13 for the manufacture of a medicament for targeting the spleen or immune cells, the medicament being for the prevention or treatment of a disease associated with immune cells.

15. Use of the LNPs according to claim 14 for the manufacture of a medicament for targeting the spleen or immune cells, the medicament being for the prevention or treatment of an autoimmune disease.

16. Use of the LNPs according to claim 15 for the manufacture of a medicament for targeting the spleen or immune cells, the active ingredient being a nucleic acid that suppresses an autoimmune response.

17. Use of the LNPs according to any one of claims 1 to 14 for the manufacture of a medicament for targeting the spleen or immune cells, the medicament being for the prevention or treatment of a T cell tumor or a B cell tumor; Preferably, the active ingredient is a nucleic acid that suppresses the growth of a T cell tumor or a B cell tumor or that kills T cell tumor cells or B cell tumor cells.

18. Use of the LNPs according to any one of claims 1 to 13 for the manufacture of a medicament for targeting the spleen or immune cells, the medicament being for the prevention or treatment of a tumor or a cancer, the active ingredient being a tumor-associated antigen or a nucleic acid encoding a tumor-associated antigen.

19. Use of the LNPs according to any one of claims 1 to 13 for the manufacture of a medicament for targeting the spleen or immune cells, the medicament being for the prevention or treatment of an infectious disease, the active ingredient being an antigen associated with an infectious disease or a nucleic acid encoding an antigen associated with an infectious disease.

20. Use of the LNPs according to any one of claims 13 to 19 for the manufacture of a medicament for targeting the spleen or immune cells, the nucleic acid being an mRNA.

21. Use of a lipid nanoparticle (LNPs) as defined in any one of claims 1 to 20 for the manufacture of a genetically edited immune cell.

22. Use of the LNPs according to claim 21 for the manufacture of a genetically edited immune cell, the genetically edited immune cell being a B cell, a T cell or an NK cell.

23. Use of the LNPs according to claim 21 or 22 for the manufacture of a genetically edited immune cell, the active ingredient being a nucleic acid required for the genetic editing.

24. Use of the LNPs of claim 23 for the preparation of a genetically edited immune cell, the active ingredient being a gRNA and a nucleic acid encoding a Cas protein.

25. Use of the LNPs of claim 24 for the preparation of a genetically edited immune cell, the active ingredient being a gRNA, a nucleic acid encoding a Cas protein and a donor DNA.

26. Use of a lipid nanoparticle (LNP) for the preparation of a modified immune cell, the LNP being as defined in any one of claims 1 to 20.

27. Use of the LNPs of claim 26 for the preparation of a modified immune cell, the modified immune cell being a CAR-T cell, a TCR-T cell or a CAR-NK cell.

28. Use of the LNPs of claim 26 or 27 for the preparation of a modified immune cell, the active ingredient being an mRNA encoding a CAR or a TCR.

29. A method of increasing the content of an active ingredient in the spleen or immune cells, the method comprising the step of contacting the LNPs or a pharmaceutical composition comprising the LNPs as defined in any one of claims 1 to 20 with the spleen or immune cells.

30. The method of increasing the content of an active ingredient in the spleen or immune cells of claim 29, the active ingredient being an mRNA.

31. The method of increasing the content of an active ingredient in the spleen or immune cells of claim 29 or 30, the method being for increasing the amount of active ingredient delivered into the spleen or immune cells.

32. A method of increasing the ratio of an active ingredient in the spleen and the liver, the method comprising the step of contacting the LNPs or a pharmaceutical composition comprising the LNPs as defined in any one of claims 1 to 20 with the spleen or immune cells.

33. The method of increasing the ratio of an active ingredient in the spleen and the liver of claim 32, the LNPs having a spleen or immune cell delivery bias, the method being for increasing the spleen / liver delivery ratio of the active ingredient.

34. The method of increasing the ratio of an active ingredient in the spleen and the liver of claim 32 or 33, the active ingredient being an mRNA.

35. The method of increasing the ratio of an active ingredient in the spleen and the liver of claim 34, the method being for increasing the expression of the mRNA in the spleen or immune cells, or the method being for increasing the spleen / liver delivery ratio of the mRNA.

36. A method of increasing expression of a nucleic acid encoding a polypeptide or protein of interest in an immune cell, the steps of the method comprising: contacting the LNPs or a pharmaceutical composition comprising the LNPs as defined in any one of claims 1 to 20 with immune cells, wherein the LNPs comprise a nucleic acid encoding a polypeptide or protein of interest.

37. The method of increasing the expression of a nucleic acid encoding a polypeptide or protein of interest in immune cells of claim 36, the immune cells being selected from one or more of the following: B cells, T cells, dendritic cells, macrophages, monocytes, NK cells and NKT cells.

38. A method of preventing or treating a disease, the steps of the method comprising: administering to a subject in need thereof LNPs as defined in any one of claims 1-20, a pharmaceutical composition comprising said LNPs, a genetically edited immune cell as defined in any one of claims 21-25, or a modified immune cell as defined in any one of claims 26-28.

39. A method for treating and / or preventing a disease associated with immune cells, the method comprising the steps of: administering to a subject in need thereof LNPs as defined in any one of claims 1-20, a pharmaceutical composition comprising said LNPs, a genetically edited immune cell as defined in any one of claims 21-25, or a modified immune cell as defined in any one of claims 26-28.

40. The method for treating and / or preventing a disease associated with immune cells according to claim 39, said disease associated with immune cells being a B cell tumor, a T cell tumor or a NK cell tumor.

41. The method for the treatment and / or prevention of a disease related to immune cells according to claim 39 or 40, which is a B-cell tumor, comprising the following steps: administering to a subject in need thereof LNPs as defined in any one of claims 1-20, a pharmaceutical composition comprising said LNPs, a genetically edited B cell as defined in any one of claims 21-25, or a modified B cell as defined in any one of claims 26-28.

42. The method for the treatment and / or prevention of a disease related to immune cells according to claim 39 or 40, which is a T cell tumor, comprising the following steps: administering to a subject in need thereof LNPs as defined in any one of claims 1-20, a pharmaceutical composition comprising said LNPs, a genetically edited T cell as defined in any one of claims 21-25, or a modified T cell as defined in any one of claims 26-28.

43. The method for the treatment and / or prevention of a disease related to immune cells, which is an NK cell tumor, according to claim 39 or 40, comprising the steps of: administering to a subject in need thereof LNPs as defined in any one of claims 1-20, a pharmaceutical composition comprising said LNPs, a genetically edited NK cell as defined in any one of claims 21-25, or a modified NK cell as defined in any one of claims 26-28.

44. A method for the treatment and / or prevention of an autoimmune disease, the method comprising: administering to a subject in need thereof LNPs as defined in any one of claims 1-20, a pharmaceutical composition comprising said LNPs, a genetically edited immune cell as defined in any one of claims 21-25, or a modified immune cell as defined in any one of claims 26-28.

45. A method for the treatment and / or prevention of a tumor (cancer), the method comprising: administering to a subject in need thereof LNPs as defined in any one of claims 1-20, a pharmaceutical composition comprising said LNPs, a genetically edited immune cell as defined in any one of claims 21-25, or a modified immune cell as defined in any one of claims 26-28.

46. A method for the treatment and / or prevention of an infectious disease, the method comprising: administering to a subject in need thereof LNPs as defined in any one of claims 1-20, a pharmaceutical composition comprising said LNPs, a genetically edited immune cell as defined in any one of claims 21-25, or a modified T cell as defined in any one of claims 26-28.

47. The method according to any one of claims 29-46, said method being performed in vivo. Preferably, said method comprises the step of administering said LNPs to a subject.

48. Lipid nanoparticles (LNPs) as defined in any one of claims 1-20.

49. The LNPs according to claim 36, said active ingredient being an mRNA.

50. A composition for making the LNPs for use of any one of claims 1-20.

51. The composition of claim 38, wherein the active ingredient is mRNA.

52. A pharmaceutical composition comprising the LNPs of claim 48.

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