Sterol-based ionized lipids and lipid nanoparticles containing the same

Abstract

Compounds, compositions and methods for delivering therapeutic, diagnostic or prophylactic agents (eg, nucleic acids) are described.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 63 / 396,125, filed August 8, 2022, the entire contents of which are incorporated herein. [Background technology]

[0002] Effective targeted delivery of active agents, such as diagnostic agents, small molecule drugs, proteins, and nucleic acids, is an ongoing medical challenge. In particular, delivery of nucleic acids into cells is made difficult by the relative instability and poor cell permeability of such species.

[0003] Lipid-containing nanoparticle compositions, liposomes, and lipoplexes have proven effective vehicles for delivering active agents to cells and / or intracellular compartments. Although various such lipid-containing nanoparticle compositions have been demonstrated, improvements in safety, efficacy, and specificity remain lacking.

[0004] Thus, there is a need to develop methods and compositions that facilitate the delivery of therapeutic, diagnostic and / or prophylactic agents, such as nucleic acids, to cells.

[0005] The compounds, compositions and methods disclosed herein address these and other needs. Summary of the Invention

[0006] The present disclosure provides compounds and uses thereof. Compositions comprising the compounds and agents of the present invention are also provided. The present disclosure also provides methods of using the compositions to deliver active agents to a subject.

[0007] In one aspect, described herein is a compound of formula Y: [ka] or a pharmaceutically acceptable salt thereof, wherein:

[0008] R 1 is H or -CH3,

[0009] [ka] represents a carbon-carbon bond or a carbon-carbon double bond,

[0010] R 7 is H or OR 16 and

[0011] R 8a and R 8b are each independently H or OR 16 or R 8a and R 8b each taken together with the atom to which it is attached forms a cycloalkyl, aryl, heterocycloalkyl, or heteroaryl;

[0012] R 9 is H, C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl;

[0013] W is CR 4a or CR 4a R 4b and if there is a double bond between W and the adjacent carbon, W is CR 4a and if there is a single bond between W and the adjacent carbon, W is CR 4a R 4b and R 4a and R 4b are each independently H, halogen, or C1-C6 alkyl;

[0014] A is [ka] and

[0015] X is O or S,

[0016] L 1-O(C=O)-, -C(=O)-, -S(O) x -, -C(=O)R a C(=O)-, -NR a C(=O)-, -NC(=O)R a C(=O)- or -C(=O)NR a C(=O)-,

[0017] L 2 is -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -S(O) x -, -SS-, -NR a C(=O)- or -C(=O)R a N-,

[0018] L 3 does not exist, -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -NR a C(=O)- or -C(=O)R a N-,

[0019] R a but [ka] wherein n is an integer from 0 to 12;

[0020] x is 0, 1, or 2,

[0021] M is CH, [ka] and

[0022] G 1 , G 2 and G 3 are independently C1 to C 12 Alkyl or C1-C 12 alkenyl,

[0023] G 4 does not exist or C1~C12 Alkyl or C1-C 12 alkenyl,

[0024] R 2 Ga-CR b R c , C6~C 24 Alkyl or C6-C 24 is alkenyl,

[0025] R 3 but [ka] and

[0026] R 6 H, OR b , CN, -C(=O)OR b , -NC(=O)R b , -C(=O)NR b , [ka] and

[0027] R b and R c are independently H, C1~C 12 Alkyl or C1-C 12 is alkenyl,

[0028] p and q are independently 0, 1, 2, 3, or 4;

[0029] R 35 but [ka] and

[0030] L 4 does not exist or [ka] and

[0031] R 10 is absent or is C1-C6 alkyl,

[0032] L 5 does not exist, or [ka] and

[0033] m is an integer of 1, 2, or 3;

[0034] L 6 does not exist or [ka] and

[0035] R 4 C3~C 10 Alkyl, C3-C 10 Cycloalkyl, C3-C 10 Alkenyl, C3-C 10 Cycloalkenyl, C3-C 10 Alkynyl, C3-C 10 aryl, C2-C9 heterocyclyl, or C2-C9 heteroaryl;

[0036] L 7 is C1-C6 alkylene,

[0037] R 13a , R 13b and R 13c are each independently C1 to C6 alkyl or C6 to C 10 is aryl,

[0038] R 14 , R 16 , R 19 , R 21 , R 22 , R 24 , R 28 , R 29 and R 31 are each independently H or C1-C6 alkyl,

[0039] R 15 -OR 16 , -NR 17 R 17 or [ka] and

[0040] R 17 are independently H, -OR 16 , C6~C 10 aryl, or C1-C6 alkyl;

[0041] R 18 are each independently halogen or C1-C6 alkyl,

[0042] o1 is an integer from 0 to 8,

[0043] p1 and p2 are each independently an integer of 0 to 2,

[0044] Z is CH2, O, S or NR 16 and

[0045] s and t are each independently 0 or 1;

[0046] R 23 is halo, hydroxyl, C1-C6 alkyl, or C1-C6 heteroalkyl;

[0047] R 20 , R 25a and R 25b , R 30a , R 30b , R 30c , R 32a , R 32b and R 34 are each independently C1 to C6 alkyl,

[0048] R 26a and R 26bare each independently H, C1-C6 alkyl, or R 26a and R 26b are combined with the atoms to which they are attached. [ka] wherein R 26c and R 26d are each independently H or substituted C1-C6 alkyl,

[0049] R 27a and R 26b are each independently H, hydroxyl, or C1-C6 alkyl;

[0050] u is 1, 2 or 3;

[0051] R 33a and R 33b are each independently C1-C6 alkyl, C1-C6 heteroalkyl, halogen, or hydroxyl;

[0052] Q is O, S or NR 16 is.

[0053] In some embodiments, provided herein are methods for delivering agents (e.g., polynucleotides). In some embodiments, provided herein are methods for diagnosing, treating, or preventing disease. In some embodiments, provided herein are methods for inducing an immune response to an infectious agent. In some embodiments, provided herein are methods for treating cancer. In some embodiments, provided herein are methods for modulating the immune system to treat cancer and other immune disorders.

[0054] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate several aspects described below. [Brief explanation of the drawings]

[0055] [Figure 1]1 shows a synthetic route for compound target 1. [Figure 2A] 1 shows the 1H NMR spectrum of compound target 1. [Figure 2B] 1 shows the mass spectrum of compound target 1. [Figure 2C] 1 shows the HPLC spectrum of compound Target 1. [Figure 3A] Characterization of SARS-CoV-2 mRNA lipid nanoparticles (LNPs). SM-102 / DVS was formulated using commercially available SM-102 lipid, and ARV-T1 was formulated using Target 1 lipid (DVS: delta mutant spike protein). Particle size and polydispersity index (PDI) of LNPs are shown. [Figure 3B] Characterization of SARS-CoV-2 mRNA lipid nanoparticles (LNPs) is shown. SM-102 / DVS is formulated using commercially available SM-102 lipids, and ARV-T1 is formulated using Target 1 lipids (DVS: delta mutant spike protein). Surface charge (zeta potential) and mRNA encapsulation efficiency of SM-102 and ARV-T1 LNPs are shown. [Figure 4] Shown is the in vitro expression of spike glycoprotein after transfection of 293T cells with 1.0 μg of DVS mRNA at 1.0 μg / mL over time. [Figure 5A] Figure 1 shows the in vitro transfection efficiency after 24 hours of 1 μg of GFP mRNA (1 μg / mL) delivered into BHK cells using LNPs. Representative fluorescence images of BHK cells after transfection are shown. [Figure 5B] Figure 1 shows the in vitro transfection efficiency of 1 μg of GFP mRNA (1 μg / mL) delivered into BHK cells using LNPs after 24 hours. Figure 2 shows the transfection efficiency analysis of GFP expression using a flow cytometer. MFI indicates mean fluorescence intensity. [Figure 6A]In vivo transfection efficiency of luciferase-expressing mRNA is shown. LNPs were formulated with the indicated ionizable lipids, and 1 μg of the formulated luciferase-expressing mRNA was injected intramuscularly. Representative images are shown 6, 24, 48, and 72 hours after administration, as determined by whole-body bioluminescence imaging using the IVIS Spectrum in vivo imaging system. [Figure 6B] Figure 1 shows the in vivo transfection efficiency of luciferase-expressing mRNA. LNPs were formulated with the indicated ionizable lipids, and 1 μg of the formulated luciferase-expressing mRNA was injected intramuscularly. Quantitative assessment of luciferase expression was determined by whole-body bioluminescence imaging using the IVIS Spectrum in vivo imaging system at 6, 24, 48, and 72 hours post-injection. [Figure 7A] Demonstrating efficient immune induction in vivo. Experimental setup: mRNA encoding the full-length spike glycoprotein of SARS-CoV-2 (δ mutant) was formulated with Target 1 (ARV-T1) lipid. Commercially available lipid SM-102 was used as a comparison. 1 μg of vaccine was injected intramuscularly as scheduled. *p<0.05, **p<0.01, ***p<0.001. [Figure 7B] Demonstrating efficient immune induction in vivo, spike-specific total IgG was assessed 14 days after the first vaccination. *p<0.05, **p<0.01, ***p<0.001. [Figure 7C] Demonstrating efficient immune induction in vivo. Spike-specific total IgG was assessed 35 days after the first vaccination. (7D) Neutralizing antibodies in serum were assessed by pseudotyped virus. (7E) Antigen-specific T cell responses were assessed by Elispot. Data are presented as mean ± SD. Statistical comparisons were performed by one-way ANOVA with Tukey's multiple comparison test. *p<0.05, **p<0.01, ***p<0.001. [Figure 7D] Demonstrating efficient immune induction in vivo. Neutralizing antibodies in serum assessed by pseudotyped virus. *p<0.05, **p<0.01, ***p<0.001. [Figure 7E] This demonstrates efficient immune induction in vivo. Antigen-specific T cell responses were assessed by Elispot. Data are presented as mean ± SD. Statistical comparisons were performed by one-way ANOVA with Tukey's multiple comparison test. *p<0.05, **p<0.01, ***p<0.001. [Figure 8A] Single-cell analysis of splenocytes from Ai14 / Cre tdTomato reporter mice demonstrates that ARV-T1-based LNPs can deliver mRNA to various cells in vivo. 10 μg of Cre mRNA formulated with ARV-T1 lipid was administered intravenously to Ai14 mice. 48 hours after injection, the percentage of CD45+ splenocytes that are tdTomato+ was determined by flow cytometry. [Figure 8B] Single-cell analysis of splenocytes from Ai14 / Cre tdTomato reporter mice demonstrates that ARV-T1-based LNPs can deliver mRNA to various cells in vivo. 10 μg of Cre mRNA formulated with ARV-T1 lipid was administered intravenously to Ai14 mice. 48 hours after injection, the distribution of CD45+tdTomato+ splenocytes was determined by flow cytometry. [Figure 9A] Figure 9 shows in vitro delivery of GFP plasmid DNA into 293T cells using ARV-T1 LNPs. Graphs depicting the size and PDI characteristics of GFP pDNA lipid nanoparticles (LNPs). (9B-9E) are images of in vivo transfection efficiency of GFP-expressing pDNA (500 ng of DNA, 48-hour incubation). (9B) 293T only, (9C) Lipo-pDNA, (9D) LNP-pDNA, and (9E) ARV-LNP-pDNA. LNP-pDNA was formulated using a conventional formulation ratio of lipids, while ARV-LNP-pDNA was formulated using a Target 1 lipid supplement containing DOTAP. [Figure 9B-9E]Figure 9 shows in vitro delivery of GFP plasmid DNA into 293T cells using ARV-T1 LNPs. Graphs depicting the size and PDI characteristics of GFP pDNA lipid nanoparticles (LNPs). (9B-9E) are images of in vivo transfection efficiency of GFP-expressing pDNA (500 ng of DNA, 48-hour incubation). (9B) 293T only, (9C) Lipo-pDNA, (9D) LNP-pDNA, and (9E) ARV-LNP-pDNA. LNP-pDNA was formulated using a conventional formulation ratio of lipids, while ARV-LNP-pDNA was formulated using a Target 1 lipid supplement containing DOTAP. [Figure 10A] Figures A–F show the in vitro delivery of small RNA (siRNA) and large RNA (saRNA) to 293T cells using ARV-T1 LNPs, respectively. (10A) is a graph showing the size and PDI characteristics of GFP siRNA lipid nanoparticles (LNPs). T1-siRNA was formulated using the same formulation as mRNA. (10B–10D) are images of in vivo delivery of GFP siRNA (siGFP) using ARV-T1 LNPs to inhibit GFP expression (10B) control, (10C) Lipo-siGFP, and (10D) ARV-T1-siGFP. (10E–10F) are images of in vitro delivery of GFP self-amplifying RNA (saRNA) using formulations of ARV-T1 LNPs, control (10E) C12-200-saRNA, and (10F) ARV-T1-saRNA. [Figures 10B-10F]Figures A–F show the in vitro delivery of small RNA (siRNA) and large RNA (saRNA) to 293T cells using ARV-T1 LNPs, respectively. (10A) is a graph showing the size and PDI characteristics of GFP siRNA lipid nanoparticles (LNPs). T1-siRNA was formulated using the same formulation as mRNA. (10B–10D) are images of in vivo delivery of GFP siRNA (siGFP) using ARV-T1 LNPs to inhibit GFP expression (10B) control, (10C) Lipo-siGFP, and (10D) ARV-T1-siGFP. (10E–10F) are images of in vitro delivery of GFP self-amplifying RNA (saRNA) using formulations of ARV-T1 LNPs, control (10E) C12-200-saRNA, and (10F) ARV-T1-saRNA. [Figure 11] 1 is a graph showing the size and PDI characteristics of formulations of the small molecule drug doxorubicin (DOX) with ARV-T1 LNPs. [Figure 12-1] 1 shows a synthetic route for compound target 11. [Figure 12-2] (As mentioned above.) [Figure 13A] 1 shows the 1H NMR spectrum of compound target 11. [Figure 13B] 1 shows the mass spectrum of compound target 11. [Figure 14A] Figure 1 shows the formulation of mRNA-LNPs using ARV-T11 lipids and the in vitro transfection efficiency of 1 μg of GFP mRNA (1 μg / mL) delivered to 293T cells using ARV-T11 LNPs after 24 hours. Figure 2 shows a graph of the size and PDI characteristics of GFP mRNA ARV-T11 LNPs. [Figures 14B-14D]Figure 14 shows the formulation of mRNA-LNPs using ARV-T11 lipids and the in vitro transfection efficiency of 1 μg of GFP mRNA (1 μg / mL) delivered to 293T cells using ARV-T11 LNPs after 24 h. (14B–14D) are representative fluorescence images of 293T cells after transfection: (14B) control, (14C) SM-102, and (14D) ARV-T11. [Figure 14E] Figure 1 shows the formulation of mRNA-LNPs using ARV-T11 lipids and the in vitro transfection efficiency of 1 μg of GFP mRNA (1 μg / mL) delivered to 293T cells using ARV-T11 LNPs after 24 hours. Figure 2 shows a graph of transfection efficiency analysis of GFP expression using a flow cytometer. [Figure 15] 1 shows a synthetic route for compound target 12. [Figure 16A] 1 shows the 1H NMR spectrum of compound target 1. [Figure 16B] 1 shows the mass spectrum of compound target 1. [Figure 16C] 1 shows the HPLC spectrum of compound Target 1. [Figure 17A] Figure 1 shows the formulation of mRNA-LNPs using ARV-T12 lipids and the in vitro transfection efficiency of 1 μg of GFP mRNA (1 μg / mL) delivered to 293T cells using ARV-T12 LNPs after 24 hours. Figure 2 shows a graph of the size and PDI characteristics of GFP mRNA ARV-T12 LNPs. [Figures 17B-17D] Figure 17 shows the formulation of mRNA-LNPs using ARV-T12 lipids and the in vitro transfection efficiency of 1 μg of GFP mRNA (1 μg / mL) delivered to 293T cells using ARV-T12 LNPs after 24 h. (17B–17D) are representative fluorescence images of 293T cells after transfection with (17B) control, (17C) SM-102, and (17D) ARV-T12. [Figure 17E]Figure 1 shows the formulation of mRNA-LNPs using ARV-T12 lipids and the in vitro transfection efficiency of 1 μg of GFP mRNA (1 μg / mL) delivered to 293T cells using ARV-T12 LNPs after 24 hours. Figure 2 shows a graph of transfection efficiency analysis of GFP expression using a flow cytometer. [Figure 18-1] 1 shows a synthetic route for compound target 13. [Figure 18-2] (As mentioned above.) [Figure 19A] 1 shows the 1H NMR spectrum of compound target 1. [Figure 19B] 1 shows the mass spectrum of compound target 1. [Figure 19C] 1 shows the HPLC spectrum of compound Target 1. [Figure 20A] Figure 1 shows the formulation of mRNA-LNPs using ARV-T13 lipids and the in vitro transfection efficiency of 1 μg of GFP mRNA (1 μg / mL) delivered to 293T cells using ARV-T13 LNPs after 24 hours. Figure 2 shows a graph of the size and PDI characteristics of GFP mRNA ARV-T13 LNPs. [Figures 20B-20D] Figure 20 shows the formulation of mRNA-LNPs using ARV-T13 lipids and the in vitro transfection efficiency of 1 μg of GFP mRNA (1 μg / mL) delivered to 293T cells using ARV-T13 LNPs after 24 h. (20B–20D) are representative fluorescence images of 293T cells after transfection with (20B) control, (20C) SM-102, and (20D) ARV-T12. [Figure 20E] Figure 1 shows the formulation of mRNA-LNPs using ARV-T13 lipids and the in vitro transfection efficiency of 1 μg of GFP mRNA (1 μg / mL) delivered to 293T cells using ARV-T13 LNPs after 24 hours. Figure 2 shows a graph of transfection efficiency analysis of GFP expression using a flow cytometer. DETAILED DESCRIPTION OF THE INVENTION

[0056] The present disclosure provides novel compounds, nanomaterials, and uses thereof. Compositions containing the compounds and drugs of the present invention are also provided. The present disclosure also provides methods of using the compositions to deliver drugs to a subject. These nanomaterials are used in applications such as gene therapy and drug delivery.

[0057] DETAILED DESCRIPTION OF THE INVENTION In the following, embodiments of the present invention will be described in detail, examples of which are illustrated in the drawings and examples. However, this invention may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

[0058] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The following definitions are provided to provide a full understanding of terms used herein.

[0059] definition General definition As used in this specification and the following claims, terms such as "comprise" (including forms such as "comprising" and "comprises," derivatives thereof, or variations thereof) and "include" (including forms such as "including" and "includes," derivatives thereof, or variations thereof) are inclusive (i.e., open-ended) and do not exclude additional elements or steps. For example, as used herein, the terms "comprises" and / or "comprising" specify the presence of stated features, integers, steps, operations, elements, and / or components, but do not exclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and / or groups thereof. Unless otherwise noted, all numbers expressing quantities of ingredients, reaction conditions, shapes, dimensions, and the like used in the specification and claims should be understood to be construed in light of, at the very least, the number of significant digits and ordinary rounding approaches, and not as an attempt to limit the application of the doctrine of equivalents to the scope of the claims.

[0060] Thus, these terms not only cover the recited element(s) or step(s), but may also include other elements or steps not expressly recited. Furthermore, as used herein, when used in conjunction with elements, the use of the terms "a," "an," and "the" can mean "one," but are also consistent with the meanings of "one or more," "at least one," and "one or more." Thus, an element preceded by "a" or "an" does not, absent further constraints, preclude the presence of additional identical elements.

[0061] Ranges may be expressed herein as from "about" one particular value and / or to "about" another particular value. "About" means within 5% of a value, e.g., within 4%, 3%, 2%, or 1% of a value. When such ranges are expressed, another aspect includes from the one particular value and / or to the other particular value. Similarly, when values are expressed as approximations by use of the antecedent "about," it is understood that the particular value forms another aspect. It is further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint. It is also understood that there are numerous values disclosed herein, and that each value is also disclosed herein as "about" that particular value in addition to the value itself. For example, if the value "10" is disclosed, then "about 10" is also disclosed. A range may be interpreted to include both the start and end of the range. For example, a range of 10% to 20% (i.e., a range of 10%-20%) may include 10%, may include 20%, and may include percentages between 10% and 20%, unless expressly stated otherwise herein.

[0062] As used herein, the terms "may," "optionally," and "optionally may" are used interchangeably and are meant to encompass not only cases where a condition occurs, but also cases where a condition does not occur. Thus, for example, a statement that a formulation "may include an excipient" is meant to encompass not only cases where the formulation includes an excipient, but also cases where the formulation does not include an excipient.

[0063] Where combinations, subsets, groups, etc. of elements are disclosed (e.g., combinations of components in a composition or combinations of steps in a method), it is understood that although specific reference may not be expressly disclosed to each of the various individual and collective combinations and permutations of those elements, each is specifically contemplated and described herein.

[0064] "Administration" to a subject includes any route of introducing or delivering an agent to a subject. Administration can be by any suitable route, such as oral, topical, transdermal, intraarticular, intraarteriolar, intradermal, intraventricular, intralesional, intranasal, rectal, vaginal, inhalation, via an implanted reservoir, or parenteral (e.g., subcutaneous, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intraspinal, intraperitoneal, intrahepatic, intralesional, and intracranial injection or infusion techniques). As used herein, "co-administration," "administration in combination," "co-administration," or "administered simultaneously" means that compounds are administered at the same time or substantially shortly after each other. In the latter case, the two compounds are administered sufficiently close in time that the results observed are indistinguishable from those achieved when the compounds are administered at the same time. "Systemic administration" refers to introducing or delivering an agent to a subject via a route that introduces or delivers the agent to a wide area of the subject's body (e.g., 50% or more of the body), for example, via entry into the circulatory or lymphatic system. In contrast, "local administration" refers to the introduction or delivery of a drug to a subject through a route that introduces or delivers the drug to the area immediately adjacent to the administration point, and does not introduce a therapeutically significant amount of the drug into the whole body.For example, a locally administered drug is easily detected in the local vicinity of the administration point, but is not detected or only detects a negligible amount in the distant part of the subject's body.Administration includes self-administration and administration by others.

[0065] As used herein, the terms "controlled release" or "controlled release drug delivery" or "sustained release" refer to the release or administration of a drug from a given dosage form in a controlled manner to achieve a desired pharmacokinetic profile in vivo. An aspect of "controlled" drug delivery is the ability to manipulate the formulation and / or dosage form to establish desired kinetics of drug release.

[0066] As used herein, the terms "beneficial agent" and "active agent" are used interchangeably to refer to a compound or composition that has a beneficial biological effect. A beneficial biological effect includes both a therapeutic effect (i.e., treatment of a disease or other undesirable physiological condition) and a prophylactic effect (i.e., prevention of a disease or other undesirable physiological condition). These terms also encompass pharmaceutically acceptable, pharmacologically active derivatives of the beneficial agents specifically referenced herein, including, but not limited to, salts, esters, amides, prodrugs, active metabolites, isomers, fragments, analogs, and the like. When the term "beneficial agent" or "active agent" is used, or when a particular agent is specifically identified, it is understood that the term includes not only the agent itself, but also pharmaceutically acceptable, pharmacologically active salts, esters, amides, prodrugs, complexes, active metabolites, isomers, fragments, analogs, and the like.

[0067] A "therapeutic agent" refers to any composition that has a beneficial biological effect. Beneficial biological effects include both therapeutic effects (e.g., treatment of a disease or other undesirable physiological condition) and prophylactic effects (e.g., prevention of a disease or other undesirable physiological condition). These terms also encompass pharmaceutically acceptable, pharmacologically active derivatives of the beneficial agents specifically referenced herein, including, but not limited to, salts, esters, amides, proagents, active metabolites, isomers, fragments, analogs, and the like. When the term "therapeutic agent" is used, or when a particular agent is specifically identified, it is understood that the term includes not only the agent itself, but also pharmaceutically acceptable, pharmacologically active salts, esters, amides, proagents, complexes, active metabolites, isomers, fragments, analogs, and the like.

[0068] "Reduction" refers to any change that results in a decrease in the amount of a symptom, disease, composition, pathology, or activity. A substance is also understood to decrease the gene output of a gene when the gene output of a gene product containing the substance is less than the output of the gene product without the substance. A reduction can also be a change in symptoms, such as fewer symptoms of a disorder than previously observed, for example. A reduction can be either an individual, median, or average decrease in a statistically significant amount of a pathology, symptom, activity, or composition. Thus, a reduction can be a 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100% reduction, as long as it is statistically significant.

[0069] "Inhibit," "inhibiting," and "inhibition" mean to decrease an activity, response, condition, disease, or other biological parameter. This includes, but is not limited to, the complete elimination of the activity, response, condition, or disease. This can further include, for example, a 10% reduction in the activity, response, condition, or disease compared to native or control levels. Thus, reduction can be a 10, 20, 30, 40, 50, 60, 70, 80, 90, 100%, or any amount of reduction in between, compared to native or control levels.

[0070] "Inactivate," "inactivating," and "inactivation" mean reducing or eliminating an activity, response, condition, disease, or other biological parameter by a chemical reaction (formation of a covalent bond) between a ligand and its biological target.

[0071] Other forms of the word "reduce" or "reducing" or "reduction" mean to lessen an event or characteristic (e.g., tumor growth). This is understood to typically be in relation to some standard or expected value, i.e., relative, but does not necessarily refer to a standard or relative value. For example, "reducing tumor growth" means reducing the rate of tumor growth compared to a standard or control.

[0072] As used herein, the term "treating" or "treatment" of a subject includes administering an agent to a subject with the intent to prevent, cure, ameliorate, alleviate, relieve, alter, cure, relieve, improve, stabilize, or affect a disease or disorder, or the symptoms of a disease or disorder. The terms "treating" and "treatment" can also refer to reducing the severity and / or frequency of symptoms, eliminating symptoms and / or underlying causes, preventing symptoms and / or underlying causes from occurring, and ameliorating or repairing damage.

[0073] Other forms of the word "prevent," such as "preventing," or "prevention," mean stopping a particular event or characteristic, stabilizing or slowing the development or progression of a particular event or characteristic, or minimizing the likelihood of a particular event or characteristic occurring. Prevention is typically more absolute than, for example, attenuation, and therefore does not require a comparison to a control. As used herein, something can be attenuated but not prevented, but something that is attenuated can also be prevented. Similarly, something can be prevented but not attenuated, but something that can be prevented can also be attenuated. When attenuation or prevention is used, it is understood that the use of other words is also expressly disclosed unless otherwise specified. For example, the terms "prevention" or "suppression" can refer to treatment that prevents or delays the onset of a disease or condition or reduces the severity of a disease or condition. Thus, while treatment can treat a disease in a subject who has symptoms of the disease, it can also prevent or suppress a disease in a subject who does not yet have some or all symptoms of the disease. As used herein, the term "preventing" a disorder or unwanted physiological event in a subject specifically refers to preventing the onset of symptoms and / or their underlying causes, and the subject may or may not have an increased susceptibility to the disorder or event.

[0074] The term "effective amount" of a therapeutic agent refers to a non-toxic, yet sufficient amount of beneficial agent to produce the desired effect. The amount of an "effective" beneficial agent will vary from subject to subject, depending on the subject's age and general condition, the particular beneficial agent, and the like. Thus, it is not always possible to specify an exact "effective amount." However, an appropriate "effective" amount for any given subject can be determined by one of ordinary skill in the art using routine experimentation. Also, as used herein, unless otherwise specified, an "effective amount" of a beneficial agent can refer to an amount that includes both a therapeutically effective amount and a prophylactically effective amount.

[0075] The "effective amount" of a drug required to achieve a therapeutic effect may vary depending on factors such as the subject's age, sex, and weight. Dosage regimens may be adjusted to provide the optimal therapeutic response. For example, the drug may be administered in divided doses daily or the dose may be proportionally reduced depending on the exigencies of the therapeutic situation.

[0076] As used herein, a "therapeutically effective amount" of a therapeutic agent refers to an amount effective to achieve a desired therapeutic result, and a "prophylactically effective amount" of a therapeutic agent refers to an amount effective to prevent an undesirable physiological condition. The therapeutically and prophylactically effective amounts of a particular therapeutic agent typically vary depending on factors such as the type and severity of the disorder or disease being treated, as well as the age, sex, and weight of the subject. The term "therapeutically effective amount" can also refer to an amount of a therapeutic agent effective to promote a desired therapeutic effect, or the rate of delivery of a therapeutic agent (e.g., amount over time). The precise desired therapeutic effect will vary depending on the condition being treated, the subject's tolerance, the agent and / or formulation being administered (e.g., potency of the therapeutic agent (drug), concentration of the agent in the formulation, etc.), and various other factors recognized by those skilled in the art.

[0077] As used herein, the term "pharmaceutically acceptable" can refer to an ingredient that is not biologically or otherwise undesirable, i.e., the ingredient, when incorporated into a pharmaceutical formulation of the present invention and administered to a subject as described herein, does not cause significant undesirable biological effects or interact in a deleterious manner with any of the other ingredients of the formulation in which it is included. When the term "pharmaceutically acceptable" is used to refer to an excipient, it generally means that the ingredient has met the necessary standards of toxicology and manufacturing testing or is included in the Inactive Ingredients Guide prepared by the U.S. Food and Drug Administration.

[0078] A "pharmaceutically acceptable carrier" (sometimes referred to as a "carrier") generally refers to a carrier or excipient useful in preparing a safe, non-toxic pharmaceutical or therapeutic composition, and includes carriers acceptable for veterinary and / or human pharmaceutical or therapeutic use. The term "carrier" or "pharmaceutically acceptable carrier" includes, but is not limited to, phosphate buffered saline, water, emulsions (such as oil / water or water / oil emulsions), and / or various types of wetting agents. As used herein, the term "carrier" encompasses, but is not limited to, any excipient, diluent, filler, salt, buffer, stabilizer, solubilizer, lipid, stabilizer, or other substance well known in the art for use in pharmaceutical formulations and further described herein.

[0079] As used herein, "pharmaceutically acceptable salts" refer to derivatives of the disclosed compounds, in which the parent compound is modified by making inorganic and organic, non-toxic acid or base addition salts. Salts of the compounds of the present invention can be synthesized from parent compounds containing a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid forms of these compounds with a stoichiometric amount of an appropriate base (such as hydroxide, carbonate, or bicarbonate of Na, Ca, Mg, or K), or by reacting the free base forms of these compounds with a stoichiometric amount of an appropriate acid. Such reactions are typically carried out in water, an organic solvent, or a mixture of the two. Generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, and acetonitrile are typical, where feasible. Salts of the compounds of the present invention further include solvates of the compounds and solvates of salts of the compounds.

[0080] Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues such as amines, alkali or organic salts of acidic residues such as carboxylic acids, etc. Pharmaceutically acceptable salts include the conventional non-toxic salts and the quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. For example, conventional non-toxic acid salts include salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, and the like; and salts prepared from organic acids such as acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, mesylic acid, esylic acid, besylic acid, sulfanilic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, isethionic acid, HOOC-(CH)-COOH (where n is 0-4), and the like, or salts prepared using a different acid that produces the same counterion. Lists of additional suitable salts are found, for example, in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., p. 1418 (1985).

[0081] Also, as used herein, the term "pharmacologically active" (or simply "active") as in "pharmacologically active" derivative or analog can refer to a derivative or analog (e.g., a salt, ester, amide, complex, metabolite, isomer, fragment, etc.) that has the same type of pharmacological activity as the parent compound, and to approximately the same extent.

[0082] A "control" is a substitute subject or sample used in an experiment for comparison purposes. Controls can be "positive" or "negative."

[0083] As used herein, "subject" means an individual. Thus, "subject" includes domestic animals (e.g., cats, dogs, etc.), livestock (e.g., cows, horses, pigs, sheep, goats, etc.), laboratory animals (e.g., mice, rabbits, rats, guinea pigs, goats, sheep, pigs, dogs, cats, etc.), and birds (e.g., chickens, turkeys, songbirds, etc.). "Subject" may further include mammals, such as primates or humans. Thus, a subject may be a human or a veterinary patient. The term "patient" refers to a subject receiving treatment by a clinician, such as a physician. The administration of a therapeutic agent may be at a dosage and for a duration effective to treat the subject. In some embodiments, the subject is a human.

[0084] As used herein, the term "nucleic acid" means a polymer composed of nucleotides, such as deoxyribonucleotides or ribonucleotides.

[0085] As used herein, the terms "ribonucleic acid" and "RNA" refer to a polymer composed of ribonucleotides.

[0086] As used herein, the terms "deoxyribonucleic acid" and "DNA" refer to a polymer composed of deoxyribonucleotides.

[0087] The term "oligonucleotide" refers to a single- or double-stranded nucleotide multimer having a length of about 2 to about 100 nucleotides. Suitable oligonucleotides can be prepared by the phosphoramidite method described in Beaucage and Carruthers, Tetrahedron Lett., 22:1859-1862 (1981), or the triester method described in Matteucci, et al., J. Am. Chem. Soc., 103:3185 (1981), both of which are incorporated herein by reference, or by other chemical methods using commercially available automated oligonucleotide synthesizers or VLSIPS™ technology. When an oligonucleotide is referred to as "double-stranded," those skilled in the art will understand that the pair of oligonucleotides typically exists in a hydrogen-bonded helical arrangement, such as that associated with DNA. In addition to 100% complementary forms of double-stranded oligonucleotides, the term "double-stranded" as used herein is also meant to refer to forms that include structural features such as bulges and loops, more detailed descriptions of which can be found in biochemistry texts such as Stryer, Biochemistry, Third Ed., (1988), which is incorporated herein by reference for all purposes.

[0088] The term "polynucleotide" refers to a single- or double-stranded polymer composed of nucleotide monomers. In some embodiments, a polynucleotide is composed of nucleotide monomers that are generally greater than 100 nucleotides in length, up to about 8,000 nucleotides or more in length.

[0089] The term "polypeptide" refers to a compound composed of a single chain of D- or L-amino acids, or a mixture of D- and L-amino acids joined by peptide bonds.

[0090] The term "complementary" refers to the topological compatibility or match of the interacting surfaces of a probe molecule and its target. Thus, the target and its probe can be described as complementary, and further, the properties of the contact surfaces are complementary to each other.

[0091] The term "hybridization" refers to the process of establishing non-covalent, sequence-specific interactions between two or more complementary nucleic acid strands to form a single hybrid, in the case of two strands, referred to as a duplex.

[0092] The term "annealing" refers to the process by which a single-stranded nucleic acid sequence pairs with a complementary sequence through hydrogen bonding to form a double-stranded nucleic acid sequence, including the reformation (renaturation) of complementary strands that have been separated by heat (thermally denatured).

[0093] The term "melting" refers to the denaturation of double-stranded nucleic acid sequences by elevated temperature, resulting in the separation of the double strand into two single strands by breaking interstrand hydrogen bonds.

[0094] The term "target" refers to a molecule that has affinity for a particular probe. Targets can be naturally occurring or artificial molecules, and can be used alone or in mixtures with other species.

[0095] The term "promoter" or "regulatory element" refers to a region or sequence determinant located upstream or downstream of the transcription start site and involved in the recognition and binding of RNA polymerase and other proteins to initiate transcription. Promoters need not be of bacterial origin; for example, promoters from viruses or other organisms can be used in the compositions, systems, or methods described herein. The term "regulatory element" is intended to include promoters, enhancers, internal ribosome entry sites (IRES), and other expression control elements (e.g., transcription termination signals such as polyadenylation signals and polyU sequences). Such regulatory elements are described, for example, in Goeddel, Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, Calif. (1990). Regulatory elements include those that direct constitutive expression of a nucleotide sequence in many types of host cells and those that direct expression of a nucleotide sequence only in specific host cells (e.g., tissue-specific regulatory sequences). Tissue-specific promoters can direct expression primarily in tissues of interest, such as muscle, neurons, bone, skin, blood, specific organs (e.g., liver, pancreas), or specific cell types (e.g., lymphocytes). Regulatory elements can direct expression in a time-dependent manner, such as cell cycle-dependent or developmental stage-dependent, which may or may not be tissue- or cell-type-specific. In some embodiments, the vector includes one or more Pol III promoters (e.g., 1, 2, 3, 4, 5, or more Pol I promoters), one or more Pol II promoters (e.g., 1, 2, 3, 4, 5, or more Pol II promoters), one or more Pol I promoters (e.g., 1, 2, 3, 4, 5, or more Pol I promoters), or a combination thereof. Examples of Pol III promoters include, but are not limited to, U6, H1, and T7 promoters.Examples of Pol II promoters include, but are not limited to, the retroviral Rous sarcoma virus (RSV) LTR promoter (optionally with the RSV enhancer), the cytomegalovirus (CMV) promoter (optionally with the CMV enhancer) [see, e.g., Boshart et al., Cell, 41:521-530 (1985)], the SV40 promoter, the dihydrofolate reductase promoter, the β-actin promoter, the phosphoglycerol kinase (PGK) promoter, the β-globin promoter, and the EF1α promoter. The term "regulatory element" encompasses enhancer elements such as WPRE, the CMV enhancer, the R-U5' segment of the HTLV-I LTR (Mol. Cell. Biol., Vol. 8(1), pp. 466-472, 1988), the SV40 enhancer, and the intron sequence between exons 2 and 3 of rabbit β-globin (Proc. Natl. Acad. Sci. USA, Vol. 78(3), pp. 1527-31, 1981). Those skilled in the art will appreciate that the design of the expression vector can depend on factors such as the choice of the host cell to be transformed and the desired expression level.

[0096] The term "recombinant" refers to a nucleic acid (e.g., polynucleotide) that has been engineered by human beings, or a copy or complement of a nucleic acid (e.g., polynucleotide) that has been engineered by human beings, or, when referring to a protein (i.e., a "recombinant protein"), the protein encoded by the recombinant nucleic acid (e.g., polynucleotide). In embodiments, a recombinant expression cassette comprising a promoter operably linked to a second nucleic acid (e.g., polynucleotide) can include a promoter that is heterologous to the second nucleic acid (e.g., polynucleotide) as a result of human engineering (e.g., by methods described in Sambrook et al., Molecular Cloning—A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, (1989) or Current Protocols in Molecular Biology Volumes 1-3, John Wiley & Sons, Inc. (1994-1998)). In another example, a recombinant expression cassette can include nucleic acids (e.g., polynucleotides) that have been combined in such a way that the nucleic acids (e.g., polynucleotides) are highly unlikely to be found in nature. For example, human-engineered restriction sites or plasmid vector sequences can flank or separate a promoter from a second nucleic acid (e.g., polynucleotide). One of skill in the art will recognize that nucleic acids (e.g., polynucleotides) can be engineered in a variety of ways, including but not limited to the above examples.

[0097] The term "expression cassette" refers to a nucleic acid construct that, upon introduction into a host cell, results in transcription and / or translation of an RNA or polypeptide, respectively. In embodiments, an expression cassette comprising a promoter operably linked to a second nucleic acid (e.g., a polynucleotide) can comprise a promoter that is heterologous to the second nucleic acid (e.g., a polynucleotide) as a result of human manipulation (e.g., by the methods described in Sambrook et al., Molecular Cloning—A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, (1989) or Current Protocols in Molecular Biology Volumes 1-3, John Wiley & Sons, Inc. (1994-1998)). In some embodiments, an expression cassette comprising a terminator (or termination sequence) operably linked to a second nucleic acid (e.g., a polynucleotide) can comprise a terminator that is heterologous to the second nucleic acid (e.g., a polynucleotide) as a result of human manipulation. In some embodiments, the expression cassette comprises a promoter operably linked to a second nucleic acid (e.g., polynucleotide) and a terminator operably linked to the second nucleic acid (e.g., polynucleotide) as a result of human engineering. In some embodiments, the expression cassette comprises an endogenous promoter. In some embodiments, the expression cassette comprises an endogenous terminator. In some embodiments, the expression cassette comprises a synthetic (or non-naturally occurring) promoter. In some embodiments, the expression cassette comprises a synthetic (or non-naturally occurring) terminator.

[0098] The terms "identical" or "percent identity" in the context of two or more nucleic acid or polypeptide sequences refer to two or more sequences or subsequences that are the same, or a specified percentage of amino acid residues or nucleotides that are the same as measured using the BLAST or BLAST 2.0 sequence comparison algorithms using the default parameters described below, or as measured by manual alignment and visual inspection (see, e.g., the NCBI website, etc.) (i.e., about 60% identity over a specified region when compared and aligned for maximum correspondence in a comparison window or specified region, preferably about 60% identity over a specified region). "Identity" refers to two or more sequences or subsequences that share 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identity. Such sequences are said to be "substantially identical." This definition also relates to and can apply to the complement of a test sequence. This definition includes sequences that have substitutions as well as deletions and / or additions. As explained below, preferred algorithms can account for gaps, etc. Preferably, identity exists over a region at least about 10 amino acids or 20 nucleotides in length, more preferably over a region 10-50 amino acids or 20-50 nucleotides in length. As used herein, percent (%) amino acid sequence identity is defined as the percentage of amino acids in a candidate sequence that are identical to those in a reference sequence after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity. Alignment for determining percent sequence identity can be accomplished in a variety of ways within the skill of the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, ALIGN-2, or Megalign (DNASTAR) software.Appropriate parameters for measuring alignment, including algorithms needed to achieve maximal alignment over the full-length of the sequences being compared, can be determined by known methods.

[0099] In sequence comparison, typically, one sequence serves as reference sequence, and test sequence is compared to it.When using sequence comparison algorithm, test sequence and reference sequence are input into computer, and subsequence coordinates are designated as needed, and sequence algorithm program parameters are designated.Preferably, default program parameters can be used, or alternative parameters can be designated.Then, sequence comparison algorithm calculates the sequence identity percentage of test sequence and reference sequence based on program parameters.

[0100] Examples of suitable algorithms for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms described in Altschul et al. (1977) Nuc. Acids Res. 25:3389-3402 and Altschul et al. (1990) J. Mol. Biol. 215:403-410, respectively. Software for performing BLAST analyses is publicly available from the National Center for Biotechnology Information (http: / / www.ncbi.nlm.nih.gov / ). This algorithm first identifies short words of length W in the query sequence and aligns them with words of the same length in the database sequences to identify high-scoring sequence pairs (HSPs) that either match or meet a certain positive score threshold T. T is referred to as the neighborhood word score threshold (Altschul et al. (1990) J. Mol. Biol. 215:403-410). These initial neighborhood word hits are used to initiate searches to find longer HSPs containing them. The word hits are extended in both directions along each sequence for as far as the cumulative alignment score can be increased. Cumulative scores for nucleotide sequences are calculated using the parameters M (reward score for a matched residue pair, always >0) and N (penalty score for mismatched residues, always <0). For amino acid sequences, a scoring matrix is used to calculate the cumulative score. Extending the word hits in each direction is stopped when the cumulative alignment score falls by an amount X from its maximum achieved value, when the accumulation of one or more negative-scoring residue alignments causes the cumulative score to fall below 0, or when the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a word length (W) of 11, an expectation (E) of 10, M=5, N=-4, and a comparison of both strands.For amino acid sequences, the BLASTP program uses as defaults a word length of 3, an expectation (E) of 10, the BLOSUM62 scoring matrix (see Henikoff and Henikoff (1989) Proc. Natl. Acad. Sci. USA 89:10915), an alignment (B) of 50, an expectation (E) of 10, M=5, N=−4, and a comparison of both strands. The BLAST algorithm also performs statistical analysis of the similarity between two sequences (see, for example, Karlin and Altschul (1993) Proc. Natl. Acad. Sci. USA 90:5873-5787). One measure of similarity provided by the BLAST algorithm is the minimum total probability (P(N)), which provides an indication of the probability that a match between two nucleotide or amino acid sequences occurs by chance. For example, if the minimum total probability in the comparison between the test nucleic acid and the reference nucleic acid is less than about 0.2, more preferably less than about 0.01, the nucleic acid is considered to be similar to the reference sequence.

[0101] The phrase "codon optimization," when referring to the coding region of a gene or nucleic acid molecule for transformation into various hosts, refers to changing the codons within the coding region of the gene or polynucleic acid molecule to reflect the general codon usage of the selected organism, without altering the polypeptide encoded by the DNA. Such optimization involves replacing at least one, or more than one, or a substantial number of codons with one or more codons that are more frequently used in the genes of the selected organism.

[0102] A nucleic acid is "operably linked" when it is placed into a functional relationship with another nucleic acid sequence. For example, DNA for a presequence or secretory leader is operably linked to DNA for a polypeptide if it is expressed as a preprotein that participates in the secretion of the polypeptide; a promoter or enhancer is operably linked to a coding sequence if it affects the transcription of the sequence; or a ribosome binding site is operably linked to a coding sequence if it is positioned so as to facilitate translation. Generally, "operably linked" means that the DNA sequences being linked are near each other, and, in the case of a secretory leader, contiguous and in reading phase. However, operably linked nucleic acids (e.g., enhancer and coding sequence) need not be contiguous. Linking is accomplished by ligation at convenient restriction sites. If such sites do not exist, synthetic oligonucleotide adapters or linkers are used in accordance with conventional practice. In embodiments, a promoter is operably linked with a coding sequence (i.e., the coding sequence is under the transcriptional control of the promoter) if it is capable of affecting expression of a protein from the coding sequence (e.g., regulating relative to the absence of the promoter).

[0103] The term "nucleobase" refers to the portion of a nucleotide that has Watson / Crick base pairing functionality. The most common natural nucleobases, adenine (A), guanine (G), uracil (U), cytosine (C), and thymine (T), have hydrogen-bonding functionality that connects one nucleic acid strand to another in a sequence-specific manner.

[0104] A nucleic acid sequence is "heterologous" to a second nucleic acid sequence if it is derived from a foreign species, or if derived from the same species but altered from its original form by human action. For example, a heterologous promoter (or heterologous 5' untranslated region (5'UTR)) operably linked to a coding sequence refers to a coding sequence in a species different from that from which the promoter was derived, or, in the same species, a coding sequence that differs from a naturally occurring allelic variant (e.g., the 5'UTR or 3'UTR of a different gene is operably linked to a nucleic acid encoding a costimulatory molecule).

[0105] The term "monoclonal antibody," as used herein, refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies within the population are identical except for natural mutations that may be present in a small subset of antibody molecules. Monoclonal antibodies herein specifically include "chimeric" antibodies in which a portion of the heavy and / or light chain is identical to or homologous to corresponding sequences in antibodies from a particular species or belonging to a particular antibody class or subclass, and the remainder of the chain(s) is identical to or homologous to corresponding sequences in antibodies from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired antagonistic activity.

[0106] The disclosed monoclonal antibodies can be made using any procedure that generates monoclonal antibodies. For example, the disclosed monoclonal antibodies can be prepared using the hybridoma method described by Kohler and Milstein, Nature, 256:495 (1975). In the hybridoma method, typically, a mouse or other suitable host animal is immunized with an immunizing agent to induce lymphocytes that produce, or are capable of producing, antibodies that specifically bind to the immunizing agent. Alternatively, lymphocytes can be immunized in vitro.

[0107] Monoclonal antibodies can also be produced by recombinant DNA methods. DNA encoding the disclosed monoclonal antibodies can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes capable of specifically binding to genes encoding the heavy and light chains of murine antibodies). Libraries of antibodies or active antibody fragments can also be generated and screened using phage display technology, as described, for example, in U.S. Patent No. 5,804,440 to Burton et al. and U.S. Patent No. 6,096,441 to Barbas et al.

[0108] In vitro methods are also suitable for preparing monovalent antibodies. Digestion of antibodies to generate fragments thereof, particularly Fab fragments, can be achieved using conventional techniques known in the art. For example, digestion can be performed using papain. Examples of papain digestion are described in WO 94 / 29348 published December 22, 1994, and U.S. Pat. No. 4,342,566. Papain digestion of antibodies typically generates two identical antigen-binding fragments, called Fab fragments, each with a single antigen-binding site, and a residual Fc fragment. Pepsin treatment generates fragments with two antigen-binding sites that are capable of cross-linking antigen.

[0109] As used herein, the term "antibody or antigen-binding fragment thereof" or "antibody or fragment thereof" includes fragments such as F(ab')2, Fab', Fab, Fv, sFv, and scFv, including chimeric and hybrid antibodies with dual or multiple antigen or epitope specificities, and hybrid fragments. Thus, fragments of antibodies that retain the ability to bind to a specific antigen are provided. For example, antibody fragments that retain binding activity are included within the meaning of the term "antibody or antigen-binding fragment thereof." Such antibodies and fragments can be produced by techniques known in the art and screened for specificity and activity according to the methods described in the Examples and general methods for producing antibodies and screening for antibody specificity and activity (see Harlow and Lane, Antibodies, A Laboratory Manual, Cold Spring Harbor Publications, New York, (1988)).

[0110] The meaning of "antibody or antigen-binding fragment thereof" also includes complexes of antibody fragments and antigen-binding proteins (single-chain antibodies). The meaning of "antibody or antigen-binding fragment thereof" also includes immunoglobulin single variable domains, such as, for example, nanobodies.

[0111] Fragments, whether attached to other sequences or not, can also contain insertions, deletions, substitutions, or other selected modifications of specific regions or specific amino acid residues, so long as the activity of the antibody or antibody fragment is not significantly altered or impaired compared to the unmodified antibody or antibody fragment. These modifications can confer additional properties, such as removing / adding amino acids capable of disulfide bonding, extending biological lifespan, or altering secretion characteristics. In any case, the antibody or antibody fragment must retain biologically active properties, such as specific binding to its cognate antigen. Functional or active regions of an antibody or antibody fragment can be identified by mutagenesis of specific regions of the protein, followed by expression and testing of the expressed polypeptide. Such methods are readily apparent to those skilled in the art and may include site-directed mutagenesis of nucleic acids encoding the antibody or antibody fragment. (Zoller, M.J. Curr. Opin. Biotechnol. 3:348-354, 1992).

[0112] As used herein, the terms "antibody" or "antibodies" can also refer to human antibodies and / or humanized antibodies. Many non-human antibodies (e.g., those derived from mice, rats, or rabbits) are naturally antigenic to humans and may provoke an undesired immune response when administered to humans. Therefore, the use of human or humanized antibodies in these methods can reduce the likelihood that antibodies administered to humans will provoke an undesired immune response.

[0113] chemical definition As used herein, the term "substituted" is contemplated to include all permissible substituents of organic compounds. In one broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. Illustrative substituents include, for example, those described below. The permissible substituents can be one or more and can be the same or different for appropriate organic compounds. For purposes of this disclosure, heteroatoms such as nitrogen can have hydrogen substituents and / or permissible substituents of organic compounds described herein that satisfy the valences of the heteroatoms. This disclosure is not intended to be limited in any manner by the permissible substituents of organic compounds. The terms "substituted" or "substituted with" also include the implicit proviso that such substitution is in accordance with the permissible valences of the substituted atom and substituent, and that the substitution results in a stable compound, e.g., a compound that does not undergo spontaneous transformation by rearrangement, cyclization, elimination, and the like.

[0114] "Z 1 "," "Z 2 "," "Z 3 " and "Z 4 " is used herein as a generic symbol to represent various specific substituents. These symbols are not limited to those disclosed herein but may be any substituent, and even if defined as one specific substituent in one instance, it may be defined as another substituent in another instance.

[0115] As used herein, the term "aliphatic" refers to a non-aromatic hydrocarbon group and includes branched and unbranched alkyl, alkenyl, or alkynyl groups.

[0116] The term "alkyl," as used herein, refers to a branched or unbranched saturated hydrocarbon group having 1 to 24 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, dodecyl, tetradecyl, hexadecyl, eicosyl, or tetracosyl. Alkyl groups can be substituted or unsubstituted. Alkyl groups can be substituted with one or more groups, including, but not limited to, alkyl, alkyl halide, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide, hydroxy, ketone, nitro, silyl, sulfoxo, sulfonyl, sulfone, sulfoxide, or thiol, as described below.

[0117] Throughout this specification, "alkyl" is used generally to refer to both unsubstituted and substituted alkyl groups, although substituted alkyl groups are also specifically referred to herein by identifying the particular substituent(s) on the alkyl group. For example, the term "halogenated alkyl" specifically refers to an alkyl group substituted with one or more halides, such as fluorine, chlorine, bromine, or iodine. The term "alkoxyalkyl" specifically refers to an alkyl group substituted with one or more alkoxy groups, as described below. The term "alkylamino" specifically refers to an alkyl group substituted with one or more amino groups, as described below, and the like. When "alkyl" is used in one instance and a specific term, such as "alkylalcohol," is used in another instance, this is not intended to imply that the term "alkyl" is not associated with the specific term, such as "alkylalcohol."

[0118] This approach is also used for other groups described herein. That is, a term such as "cycloalkyl" refers to both unsubstituted and substituted cycloalkyl moieties, but substituted moieties are also specifically identified herein; for example, a particular substituted cycloalkyl may be referred to specifically as an "alkylcycloalkyl," etc. Similarly, a substituted alkoxy may be specifically referred to as, for example, a "halogenated alkoxy," a particular substituted alkenyl may be specifically referred to as, for example, an "alkenylalcohol," etc. Again, the practice of using a general term such as "cycloalkyl" and a specific term such as "alkylcycloalkyl" does not mean that the general term does not include the specific term.

[0119] The term "alkoxy" as used herein refers to an alkyl group attached through a single terminal ether linkage, i.e., an "alkoxy" group is Z 1 -OZ is alkyl as defined above 1 It can be defined as:

[0120] The term "alkenyl," as used herein, refers to a hydrocarbon group of 2 to 24 carbon atoms with a structural formula containing at least one carbon-carbon double bond. 1 Z 2 Asymmetric structures such as C=C(ZZ) are intended to include both the E and Z isomers. This can be inferred in structural formulas herein where an asymmetric alkene is present, or can be explicitly shown by the bond symbol C=C. Alkyl groups can be substituted with one or more groups, including, but not limited to, alkyl, alkyl halide, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide, hydroxy, ketone, nitro, silyl, sulfoxo, sulfonyl, sulfone, sulfoxide, or thiol, as described below.

[0121] The term "alkynyl," as used herein, refers to a hydrocarbon group of 2 to 24 carbon atoms having a structural formula containing at least one carbon-carbon triple bond. The alkynyl group may be substituted with one or more groups, including, but not limited to, alkyl, alkyl halide, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide, hydroxy, ketone, nitro, silyl, sulfoxo, sulfonyl, sulfone, sulfoxide, or thiol, as described below.

[0122] The term "aryl," as used herein, refers to a group containing any carbon-based aromatic group, including, but not limited to, benzene, naphthalene, phenyl, biphenyl, phenoxybenzene, and the like. The term "heteroaryl" is defined as a group containing an aromatic group with at least one heteroatom incorporated within the ring of the aromatic group. Examples of heteroatoms include, but are not limited to, nitrogen, oxygen, sulfur, phosphorus, and the like. The term "non-heteroaryl," included within the term "aryl," defines a group containing an aromatic group without a heteroatom. An aryl or heteroaryl group may be substituted or unsubstituted. An aryl or heteroaryl group may be substituted with one or more groups, including, but not limited to, alkyl, alkyl halide, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide, hydroxy, ketone, nitro, silyl, sulfoxo, sulfonyl, sulfone, sulfoxide, or thiol, as described herein. The term "biaryl" is a specific type of aryl group and is included within the definition of aryl. Biaryl refers to two aryl groups joined through a fused ring structure, as in naphthalene, or two aryl groups joined through one or more carbon-carbon bonds, as in biphenyl.

[0123] The term "cycloalkyl," as used herein, refers to a non-aromatic carbon-based ring composed of at least three carbon atoms. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like. The term "heterocycloalkyl" refers to a cycloalkyl group, as defined above, in which at least one of the ring carbon atoms is replaced with a heteroatom, such as nitrogen, oxygen, sulfur, or phosphorus. Cycloalkyl and heterocycloalkyl groups can be substituted or unsubstituted. Cycloalkyl and heterocycloalkyl groups can be substituted with one or more groups, including, but not limited to, alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide, hydroxy, ketone, nitro, silyl, sulfoxo, sulfonyl, sulfone, sulfoxide, or thiol, as described herein.

[0124] The term "cycloalkenyl," as used herein, refers to a non-aromatic carbon-based ring composed of at least three carbon atoms and containing at least one double bond, i.e., C=C. Examples of cycloalkenyl groups include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, and the like. The term "heterocycloalkenyl" is a type of cycloalkenyl group, as defined above, and is included in the meaning of the term "cycloalkenyl," in which at least one of the ring carbon atoms is replaced with a heteroatom, such as nitrogen, oxygen, sulfur, or phosphorus. Cycloalkenyl and heterocycloalkenyl groups can be substituted or unsubstituted. The cycloalkenyl and heterocycloalkenyl groups may be substituted with one or more groups described herein, including, but not limited to, alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide, hydroxy, ketone, nitro, silyl, sulfoxo, sulfonyl, sulfone, sulfoxide, or thiol.

[0125] As used herein, the term "cyclic group" refers to either aryl groups, non-aryl groups (i.e., cycloalkyl, heterocycloalkyl, cycloalkenyl, and heterocycloalkenyl groups), or both. A cyclic group has one or more ring systems, which may be substituted or unsubstituted. A cyclic group may contain one or more aryl groups, one or more non-aryl groups, or one or more aryl groups and one or more non-aryl groups.

[0126] The term "aldehyde" as used herein is represented by the formula -C(O)H. Throughout this specification, "C(O)" or "CO" are shorthand notations for C=O.

[0127] As used herein, the term "amine" or "amino" refers to a group of the formula -NZ 1 Z 2 wherein Z and Z can each be a substituent described herein, for example, a hydrogen, alkyl, halogenated alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl, or heterocycloalkenyl group as described above.

[0128] The term "carboxylic acid" as used herein is represented by the formula -C(O)OH. A "carboxylate" or "carboxyl" group as used herein is represented by the formula -C(O)O-.

[0129] As used herein, the term "ester" refers to an ester of the formula -OC(O)Z 1 or -C(O)OZ 1 wherein Z1 can be an alkyl, halogenated alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl, or heterocycloalkenyl group as described above.

[0130] As used herein, the term "ether" refers to a group of the formula Z 1 OZ 2 In the formula, Z 1 and Z2 can independently be an alkyl, halogenated alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl, or heterocycloalkenyl group as described above.

[0131] As used herein, the term "ketone" refers to a compound of formula Z 1 C(O)Z 2 In the formula, Z 1 and Z 2 can independently be an alkyl, halogenated alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl, or heterocycloalkenyl group as described above.

[0132] As used herein, the term "halide" or "halogen" refers to fluorine, chlorine, bromine and iodine.

[0133] The term "hydroxyl" as used herein is represented by the formula --OH.

[0134] The term "nitro" as used herein is represented by the formula -NO2.

[0135] As used herein, the term "silyl" refers to a group of the formula -SiZ 1 Z 2 Z 3 In the formula, Z 1 , Z 2 and Z 3 can independently be hydrogen, alkyl, halogenated alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl, or heterocycloalkenyl groups as described above.

[0136] As used herein, the term "sulfonyl" refers to a group of the formula -S(O)Z 1wherein Z1 can be hydrogen, alkyl, halogenated alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl, or heterocycloalkenyl group as described above.

[0137] The term "sulfonylamino" or "sulfonamide" as used herein is represented by the formula -S(O)2NH-.

[0138] As used herein, the term "phosphonyl" refers to a group of the formula -P(O)(OZ 1 )2, wherein Z1 can be hydrogen, alkyl, halogenated alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl, or heterocycloalkenyl group as described above.

[0139] The term "thiol" as used herein is represented by the formula --SH.

[0140] The term "thio" as used herein is represented by the formula -S-.

[0141] As used herein, "R 1 "," "R 2 "," "R 3 "," "R n ", etc. (n is an integer) can independently have one or more of the above groups. For example, if R1 is a straight chain alkyl group, one of the hydrogen atoms of the alkyl group can be optionally replaced with a hydroxyl group, an alkoxyl group, an amine group, an alkyl group, a halide, etc. Depending on the group selected, the first group can be incorporated within the second group, or the first group can be pendant (i.e., attached) to the second group. For example, in the phrase "an alkyl group comprising an amino group," the amino group can be incorporated within the backbone of the alkyl group. Alternatively, the amino group can be attached to the backbone of the alkyl group. The nature of the group(s) selected will determine whether the first group is embedded in or attached to the second group.

[0142] As used herein, "ionizable lipid" refers to a lipid that has a group or atom that can be charged (ionized) depending on the pH of the environment. Typically, the charge of primary, secondary, and tertiary amines depends on the pH of the system. At low pH levels, these amines tend to be strongly cationic. At high pH levels, these amines do not ionize.

[0143] Unless otherwise stated, formulas with chemical bonds shown only with solid lines, rather than wedges or dashed lines, contemplate all possible isomers, such as individual enantiomers, diastereomers, meso compounds, and mixtures of isomers, such as racemic and scalemic mixtures.

[0144] Reference will now be made in detail to certain embodiments of the disclosed materials, compounds, compositions, articles and methods, examples of which are illustrated in the accompanying examples and drawings.

[0145] compound Described herein are compounds of formula Y: [ka] or a pharmaceutically acceptable salt thereof, wherein:

[0146] R 1 is H or -CH3,

[0147] [ka] represents a carbon-carbon bond or a carbon-carbon double bond,

[0148] R 7 is H or OR 16 and

[0149] R 8a and R 8b are each independently H or OR 16 or R8a and R 8b each taken together with the atom to which it is attached forms a cycloalkyl, aryl, heterocycloalkyl, or heteroaryl;

[0150] R 9 is H, C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl;

[0151] W is CR 4a or CR 4a R 4b and if there is a double bond between W and the adjacent carbon, W is CR 4a and if there is a single bond between W and the adjacent carbon, W is CR 4a R 4b and R 4a and R 4b are each independently H, halogen, or C1-C6 alkyl;

[0152] A is [ka] and

[0153] X is O or S,

[0154] L 1 -O(C=O)-, -C(=O)-, -S(O) x -, -C(=O)R a C(=O)-, -NR a C(=O)-, -NC(=O)R a C(=O)- or -C(=O)NR a C(=O)-,

[0155] L 2 is -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -S(O) x -, -SS-, -NR a C(=O)- or -C(=O)R a N-,

[0156] L 3 does not exist, -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -NR a C(=O)- or -C(=O)R a N-,

[0157] R a but [ka] wherein n is an integer from 0 to 12;

[0158] x is 0, 1, or 2,

[0159] M is CH, [ka] and

[0160] G 1 , G 2 and G 3 are independently C1 to C 12 Alkyl or C1-C 12 alkenyl,

[0161] G 4 does not exist or C1~C 12 Alkyl or C1-C 12 alkenyl,

[0162] R 2 Ga-CR b R c , C6~C 24 Alkyl or C6-C 24 is alkenyl,

[0163] R 3 but [ka] and

[0164] R6 H, OR b , CN, -C(=O)OR b , -NC(=O)R b , -C(=O)NR b , [ka] and

[0165] R b and R c are independently H, C1~C 12 Alkyl or C1-C 12 is alkenyl,

[0166] p and q are independently 0, 1, 2, 3, or 4;

[0167] R 35 but [ka] and

[0168] L 4 does not exist or [ka] and

[0169] R 10 is absent or is C1-C6 alkyl,

[0170] L 5 does not exist, or [ka] and

[0171] m is an integer of 1, 2, or 3;

[0172] L 6 does not exist or [ka] and

[0173] R 4 C3~C 10 Alkyl, C3-C 10 Cycloalkyl, C3-C 10 Alkenyl, C3-C 10 Cycloalkenyl, C3-C 10 Alkynyl, C3-C 10 aryl, C2-C9 heterocyclyl, or C2-C9 heteroaryl;

[0174] L 7 is C1-C6 alkylene,

[0175] R 13a , R 13b and R 13c are each independently C1 to C6 alkyl or C6 to C 10 is aryl,

[0176] R 14 , R 16 , R 19 , R 21 , R 22 , R 24 , R 28 , R 29 and R 31 are each independently H or C1-C6 alkyl,

[0177] R 15 -OR 16 , -NR 17 R 17 or [ka] and

[0178] R 17 are independently H, -OR 16 , C6~C 10 aryl, or C1-C6 alkyl;

[0179] R 18are each independently halogen or C1-C6 alkyl,

[0180] o1 is an integer from 0 to 8,

[0181] p1 and p2 are each independently an integer of 0 to 2,

[0182] Z is CH2, O, S or NR 16 and

[0183] s and t are each independently 0 or 1;

[0184] R 23 is halo, hydroxyl, C1-C6 alkyl, or C1-C6 heteroalkyl;

[0185] R 20 , R 25a and R 25b , R 30a , R 30b , R 30c , R 32a , R 32b and R 34 are each independently C1 to C6 alkyl,

[0186] R 26a and R 26b are each independently H, C1-C6 alkyl, or R 26a and R 26b are combined with the atoms to which they are attached. [ka] wherein R 26c and R 26d are each independently H or substituted C1-C6 alkyl,

[0187] R 27a and R 26b are each independently H, hydroxyl, or C1-C6 alkyl;

[0188] u is 1, 2 or 3;

[0189] R 33a and R 33b are each independently C1-C6 alkyl, C1-C6 heteroalkyl, halogen, or hydroxyl;

[0190] Q is O, S or NR 16 is.

[0191] In some embodiments, R 35 but [ka] wherein:

[0192] L 4 does not exist or [ka] and

[0193] R 10 is absent or is C1-C6 alkyl,

[0194] L 5 does not exist, or [ka] and

[0195] m is an integer of 1, 2, or 3;

[0196] L 6 does not exist or [ka] and

[0197] R 4 C3~C 10 Alkyl, C3-C 10 Cycloalkyl, C3-C 10Alkenyl, C3-C 10 Cycloalkenyl, C3-C 10 Alkynyl, C3-C 10 It is aryl, C2-C9 heterocyclyl, or C2-C9 heteroaryl.

[0198] In one aspect, described herein is a compound of formula I: [ka] or a pharmaceutically acceptable salt thereof, wherein:

[0199] R 1 is H or -CH3,

[0200] [ka] represents a carbon-carbon bond or a carbon-carbon double bond,

[0201] L 4 does not exist or [ka] and

[0202] R 10 is absent or is C1-C6 alkyl,

[0203] L 5 does not exist, or [ka] and

[0204] m is an integer of 1, 2, or 3;

[0205] L 6 does not exist or [ka] and

[0206] R 4 C3~C 10 Alkyl, C3-C 10 Cycloalkyl, C3-C 10 Alkenyl, C3-C 10 Cycloalkenyl, C3-C 10 Alkynyl, C3-C 10 aryl, C2-C9 heterocyclyl, C2-C9 heteroaryl;

[0207] R 7 is H or OR 16 and

[0208] R 8a and R 8b are each independently H or OR 16 or R 8a and R 8b each taken together with the atom to which it is attached forms a cycloalkyl, aryl, heterocycloalkyl, or heteroaryl;

[0209] R 16 is H or C1-C6 alkyl,

[0210] R 9 is H, C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl;

[0211] W is CR 4a or CR 4a R 4b and if there is a double bond between W and the adjacent carbon, W is CR 4a and if there is a single bond between W and the adjacent carbon, W is CR 4a R 4b and R 4a and R 4b are each independently H, halogen, or C1-C6 alkyl;

[0212] A is [ka] and

[0213] X is O or S,

[0214] L 1 -O(C=O)-, -C(=O)-, -S(O) x -, -C(=O)R a C(=O)-, -NR a C(=O)-, -NC(=O)R a C(=O)- or -C(=O)NR a C(=O)-,

[0215] L 2 is -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -S(O) x -, -SS-, -NR a C(=O)- or -C(=O)R a N-,

[0216] L 3 does not exist, -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -NR a C(=O)- or -C(=O)R a N-,

[0217] R a but [ka] wherein n is an integer from 0 to 12;

[0218] x is 0, 1, or 2,

[0219] M is CH, [ka] and

[0220] G 1 , G 2 and G 3 are independently C1 to C12 Alkyl or C1-C 12 alkenyl,

[0221] G 4 does not exist or C1~C 12 Alkyl or C1-C 12 alkenyl,

[0222] R 2 Ga-CR b R c , C6~C 24 Alkyl or C6-C 24 is alkenyl,

[0223] R 3 but [ka] and

[0224] R 6 H, OR b , CN, -C(=O)OR b , -NC(=O)R b , -C(=O)NR b , [ka] and

[0225] R b and R c are independently H, C1~C 12 Alkyl or C1-C 12 alkenyl, and

[0226] p and q are independently 0, 1, 2, 3, or 4.

[0227] In some embodiments, described herein are compounds of formula Ia: [ka] or a pharmaceutically acceptable salt thereof, wherein:

[0228] R 1 is H or -CH3,

[0229] [ka] represents a carbon-carbon bond or a carbon-carbon double bond,

[0230] L 4 does not exist or [ka] and

[0231] [ka] L 5 does not exist, or and

[0232] wherein m is an integer of 1, 2, or 3;

[0233] L 6 does not exist or [ka] and

[0234] R 4 C3~C 10 Alkyl, C3-C 10 Cycloalkyl, C3-C 10 Alkenyl, C3-C 10 Cycloalkenyl, C3-C 10 Alkynyl, C3-C 10 aryl, C2-C9 heterocyclyl, C2-C9 heteroaryl;

[0235] R 10 is absent or is C1-C6 alkyl,

[0236] A is [ka] and

[0237] X is O or S,

[0238] L 1 -O(C=O)-, -C(=O)-, -S(O) x -, -C(=O)R a C(=O)-, -NR a C(=O)-, -NC(=O)R a C(=O)- or -C(=O)NR a C(=O)-,

[0239] L 2 is -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -S(O) x -, -SS-, -NR a C(=O)- or -C(=O)R a N-,

[0240] L 3 does not exist, -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -NR a C(=O)- or -C(=O)R a N-,

[0241] R a but [ka] wherein n is an integer from 0 to 12;

[0242] x is 0, 1, or 2, M is CH, [ka] and

[0243] G 1 , G 2 and G 3are independently C1 to C 12 Alkyl or C1-C 12 alkenyl,

[0244] G 4 does not exist or C1~C 12 Alkyl or C1-C 12 alkenyl,

[0245] R 2 Ga-CR b R c , C6~C 24 Alkyl or C6-C 24 is alkenyl,

[0246] R 3 but [ka] and

[0247] R 6 H, OR b , CN, -C(=O)OR b , -NC(=O)R b , -C(=O)NR b , [ka] and

[0248] R b and R c are independently H, C1~C 12 Alkyl or C1-C 12 alkenyl, and

[0249] p and q are independently 0, 1, 2, 3, or 4.

[0250] In some embodiments, the compound of formula Y is [ka] or a pharmaceutically acceptable salt thereof,

[0251] In the formula, A, R 1 , R 7 , R 8a , R 8b , R 9 , the same as when W is the formula Y.

[0252] In some embodiments, the compound of formula Y is [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] or a pharmaceutically acceptable salt thereof, In the formula, A is the same as in formula Y.

[0253] In some embodiments, described herein are compounds of Formula II or Formula III: [ka] [ka] or a pharmaceutically acceptable salt thereof, wherein:

[0254] R 1 is H or -CH3,

[0255] X is O or S,

[0256] [ka] represents a carbon-carbon bond or a carbon-carbon double bond,

[0257] L 4 Does not exist [ka] and

[0258] R 10 is absent or is C1-C6 alkyl,

[0259] L 5 does not exist, or [ka] and

[0260] m is an integer of 1, 2, or 3;

[0261] L 6 does not exist or [ka] and

[0262] R 4 C3~C 10 Alkyl, C3-C 10 Cycloalkyl, C3-C 10 Alkenyl, C3-C 10Cycloalkenyl, C3-C 10 alkynyl, or derivatives thereof,

[0263] L 1 -O(C=O)-, -C(=O)-, -S(O) x -, -C(=O)R a C(=O)-, -NR a C(=O)-, -NC(=O)R a C(=O)- or -C(=O)NR a C(=O)-,

[0264] L 2 is -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -S(O) x -, -SS-, -NR a C(=O)- or -C(=O)R a N-,

[0265] L 3 does not exist, -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -NR a C(=O)- or -C(=O)R a N-,

[0266] R a but [ka] wherein n is an integer from 0 to 12;

[0267] x is 0, 1, or 2,

[0268] M is CH, [ka] and

[0269] G 1 , G 2 and G 3 are independently C1 to C 12 Alkyl or C1-C12 alkenyl,

[0270] G 4 does not exist or C1~C 12 Alkyl or C1-C 12 alkenyl,

[0271] R 2 Ga-CR b R c , C6~C 24 Alkyl or C6-C 24 is alkenyl,

[0272] R 3 but [ka] and

[0273] R 6 H, OR b , CN, -C(=O)OR b , -NC(=O)R b , -C(=O)NR b , [ka] and

[0274] R b and R c are independently H, C1~C 12 Alkyl or C1-C 12 alkenyl, and

[0275] p and q are independently 0, 1, 2, 3, or 4.

[0276] In some embodiments, R 1 In some embodiments, X is O. In some embodiments, L 4 but [ka] In some embodiments, L 5 but [ka] In some embodiments, L 5 but [ka] m is 3. In some embodiments, L 6 In some embodiments, R 4 is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, pentyl, hexyl, heptyl, octyl, nonyl, or decyl. 4 is isopropyl. In some embodiments, M is CH.

[0277] In some embodiments, R 3 but [ka] is.

[0278] In some embodiments, R 6 H, OR b , CN, -C(=O)OR b , -NC(=O)R b , -C(=O)NR b , [ka] is.

[0279] In some embodiments, described herein are compounds of Formula IV or Formula V: [ka] [ka] or a pharmaceutically acceptable salt thereof, wherein:

[0280] L 1 -O(C=O)-, -C(=O)-, -S(O) x -, -C(=O)R a C(=O)-, -NR a C(=O)-, -NC(=O)R a C(=O)- or -C(=O)NR a C(=O)-,

[0281] L 2 is -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -S(O) x -, -SS-, -NR a C(=O)- or -C(=O)R a N-,

[0282] L 3 does not exist, -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -NR a C(=O)- or -C(=O)R a N-,

[0283] R a but [ka] wherein n is an integer from 0 to 12;

[0284] x is 0, 1, or 2,

[0285] M is CH, [ka] and

[0286] G 1 , G 2 and G 3 are independently C1 to C 12 Alkyl or C1-C 12 alkenyl,

[0287] G 4 does not exist or C1~C 12 Alkyl or C1-C 12 alkenyl,

[0288] R 2 Ga-CR b R c , C6~C 24 Alkyl or C6-C 24 is alkenyl,

[0289] R 3 but [ka] and

[0290] R 6 H, OR b , CN, -C(=O)OR b , -NC(=O)R b , -C(=O)NR b , [ka] and

[0291] R b and R c are independently H, C1~C 12 Alkyl or C1-C 12 alkenyl, and

[0292] p and q are independently 0, 1, 2, 3, or 4.

[0293] In some embodiments, L 1 is —C(═O)—. In some embodiments, L 2 is —(C═O)O—. In some embodiments, L 3 In some embodiments, G 1 is a C5 alkyl. In some embodiments, G 2is C alkyl. In some embodiments, G 3 is C alkyl. In some embodiments, G 4 does not exist.

[0294] In some embodiments, R 2 Ga-CR b R c In some embodiments, R b and R c is C alkyl. In some embodiments, R b and R c is C4 alkyl. In some embodiments, R b and R c is C alkyl. In some embodiments, R b and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b and R c C 10 In some embodiments, R 2 Ga-CR b R c and R b and R c C 12 In some embodiments, R 2 Ga-CR b R c and R b C 12 alkyl, and R c C 10 In some embodiments, R 2 Ga-CR b R c and R b C 10 alkyl, and R c C 12 In some embodiments, R 2 Ga-CR b R c and R b is C8 alkyl, and R c C10 In some embodiments, R 2 Ga-CR b R c and R b is C6 alkyl, and R c C 10 In some embodiments, R 2 Ga-CR b R c and R b is a C4 alkyl, and R c C 10 In some embodiments, R 2 Ga-CR b R c and R b is a C2 alkyl, and R c C 10 In some embodiments, R 2 Ga-CR b R c and R b is C1 alkyl, and R c C 10 In some embodiments, R 2 Ga-CR b R c and R b C 10 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b C 10 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b C 10 alkyl, and R c is C4 alkyl. In some embodiments, R 2 Ga-CR b R c and R b C 10 alkyl, and R cis C alkyl. In some embodiments, R 2 Ga-CR b R c and R b C 10 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C8 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C6 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is a C4 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is a C2 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C1 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C8 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C8 alkyl, and R c is C4 alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C8 alkyl, and Rc is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C8 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C8 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C6 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is a C4 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is a C2 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C1 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C6 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C6 alkyl, and R c is C4 alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C6 alkyl, and Rc is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C6 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C8 alkyl, and R c is C4 alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C6 alkyl, and R c is C4 alkyl. In some embodiments, R 2 Ga-CR b R c and R b is a C4 alkyl, and R c is C4 alkyl. In some embodiments, R 2 Ga-CR b R c and R b is a C2 alkyl, and R c is C4 alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C1 alkyl, and R c is C4 alkyl. In some embodiments, R 2 Ga-CR b R c and R b is a C4 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is a C4 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is a C4 alkyl, and Rc is C4 alkyl. In some embodiments, R 2 Ga-CR b R c and R b is a C4 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is a C4 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C8 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C6 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is a C4 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is a C2 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C1 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is a C2 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is a C2 alkyl, and Rc is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is a C2 alkyl, and R c is C4 alkyl. In some embodiments, R 2 Ga-CR b R c and R b is a C2 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C8 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C6 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is a C4 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is a C2 alkyl, and R c C 1 alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C1 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C1 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R bis C1 alkyl, and R c is C alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C1 alkyl, and R c is C4 alkyl. In some embodiments, R 2 Ga-CR b R c and R b is C1 alkyl, and R c is a C2 alkyl.

[0295] In some embodiments, R 2 Ga-CR b R c In some embodiments, R b and R c is C alkenyl. In some embodiments, R b and R c is C4 alkenyl. In some embodiments, R b and R c is C alkenyl. In some embodiments, R b and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b and R c C 10 In some embodiments, R is alkenyl. 2 Ga-CR b R c and R b and R c C 12 In some embodiments, R is alkenyl. 2 Ga-CR b R c and R b C 12 alkenyl, and R c C 10 In some embodiments, R is alkenyl. 2 Ga-CR b R cand R b C 10 alkenyl, and R c C 12 In some embodiments, R is alkenyl. 2 Ga-CR b R c and R b is C8 alkenyl, and R c C 10 In some embodiments, R is alkenyl. 2 Ga-CR b R c and R b is C6 alkenyl, and R c C 10 In some embodiments, R is alkenyl. 2 Ga-CR b R c and R b is C4 alkenyl, and R c C 10 In some embodiments, R is alkenyl. 2 Ga-CR b R c and R b is C2 alkenyl, and R c C 10 In some embodiments, R is alkenyl. 2 Ga-CR b R c and R b is C1 alkenyl, and R c C 10 In some embodiments, R is alkenyl. 2 Ga-CR b R c and R b C 10 and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b C 10 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b Rc and R b C 10 alkenyl, and R c is C4 alkenyl. In some embodiments, R 2 Ga-CR b R c and R b C 10 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b C 10 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C8 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C6 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C4 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C2 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C1 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C8 alkenyl, and R cis C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C8 alkenyl, and R c is C4 alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C8 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C8 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C8 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C6 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C4 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C2 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C1 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C6 and Rc is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C6 alkenyl, and R c is C4 alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C6 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C6 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C8 alkenyl, and R c is C4 alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C6 alkenyl, and R c is C4 alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C4 alkenyl, and R c is C4 alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C2 alkenyl, and R c is C4 alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C1 alkenyl, and R c is C4 alkenyl. In some embodiments, R 2 Ga-CR b R c and R bis C4 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C4 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C4 alkenyl, and R c is C4 alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C4 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C4 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C8 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C6 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C4 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C2 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R cand R b is C1 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C2 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C2 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C2 alkenyl, and R c is C4 alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C2 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C8 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C6 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C4 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C2 alkenyl, and Rc is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C1 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C1 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C1 alkenyl, and R c is C alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C1 alkenyl, and R c is C4 alkenyl. In some embodiments, R 2 Ga-CR b R c and R b is C1 alkenyl, and R c is a C2 alkenyl.

[0296] In some embodiments, R2 is -CH 13 , -CH 15 , -CH 17 , -C 10 H 21 , -C 11 H 23 , -C 12 H 25 , -C 13 H 27 , -C 14 H 29 , -C 15 H 31 , -C 16 H 33 , -C 17 H 35 , -C 18 H 36 , -C 19 H 39 , -C20 H 41 , -C 21 H 43 , -C 22 H 45 , -C 23 H 47 , -C 24 H 49 , -CH 12 , -CH 14 , -CH 16 , -C 10 H 20 , -C 11 H 21 , -C 12 H 24 , -C 13 H 26 , -C 14 H 28 , -C 15 H 30 , -C 16 H 32 , -C 17 H 34 , -C 18 H 35 , -C 19 H 38 , -C 20 H 40 , -C 21 H 42 , -C 22 H 44 , -C 23 H 46 , or -C 24 H 48 is.

[0297] In some embodiments, R 3 but [ka] In some embodiments, p and q are 0. In some embodiments, p and q are 1. In some embodiments, p and q are 2. In some embodiments, p and q are 3. In some embodiments, p and q are 4. In some embodiments, p is 0 and q is 1. In some embodiments, p is 0 and q is 2. In some embodiments, p is 0 and q is 3. In some embodiments, p is 0 and q is 4. In some embodiments, p is 1 and q is 2. In some embodiments, p is 1 and q is 3. In some embodiments, p is 1 and q is 4. In some embodiments, p is 2 and q is 1. In some embodiments, p is 2 and q is 3. In some embodiments, p is 2 and q is 4. In some embodiments, p is 3 and q is 1. In some embodiments, p is 3 and q is 2. In some embodiments, p is 3 and q is 4. In some embodiments, p is 4 and q is 1. In some embodiments, p is 4 and q is 2. In some embodiments, p is 4 and q is 3.

[0298] In some embodiments, M is CH. In some embodiments, M is [ka] In some embodiments, M is [ka] In some embodiments, M is [ka] In some embodiments, M is [ka] In some embodiments, M is [ka] In some embodiments, M is [ka] In some embodiments, M is [ka] is.

[0299] In some embodiments, R 3 but [ka] is.

[0300] In some embodiments, R 6 H, OR b , CN, -C(=O)OR b , -NC(=O)R b , -C(=O)NR b , [ka] is.

[0301] In some embodiments, the compound of formula Y, I, Ia, or II-V is [ka] [ka] [ka] [ka] [ka] [ka] [ka] or a pharmaceutically acceptable salt thereof.

[0302] In some embodiments, the compound of formula Y, I, Ia, or II-V is [ka] or a pharmaceutically acceptable salt thereof.

[0303] In some embodiments, the compound of formula Y, I, Ia, or II-V is [ka] or a pharmaceutically acceptable salt thereof, or any combination thereof.

[0304] In some embodiments, the compound of formula Y, I, Ia, or II-V is [ka] It could be.

[0305] In some embodiments, the compound of formula Y, I, Ia, or II-V is [ka] It could be.

[0306] In some embodiments, the compound of formula Y, I, Ia, or II-V is [ka] or a pharmaceutically acceptable salt thereof.

[0307] In some embodiments, the compound of formula Y, I, Ia, or II-V is [ka] It could be.

[0308] In some embodiments, the compound of formula Y, I, Ia, or II-V is [ka] or a pharmaceutically acceptable salt thereof.

[0309] Described herein are compositions comprising a compound of Formula Y, I, Ia, II, III, IV, aV, or any combination thereof, and an active agent.

[0310] composition The compositions described herein comprise a compound of Formula Y, I, Ia, II, III, IV, or V, or any combination thereof, and an active agent. The compositions may further comprise any excipients that may be useful for various medical and non-medical applications. For example, the pharmaceutical compositions described herein may be useful for delivering an effective amount of a drug to a subject in need thereof. The compositions described herein may be useful for non-medical applications, such as emulsions or emulsifiers useful for food ingredients, fire suppression, surface disinfection, oil removal, etc.

[0311] In some embodiments, the composition may be a lipid nanoparticle dispersion, a liposomal formulation, a lipid emulsion, or any combination thereof comprising a compound of Formula Y, I, Ia, II, III, IV, V, or any combination thereof.

[0312] In one aspect, the present disclosure provides a composition comprising lipid nanoparticles comprising a compound of Formula Y, I, Ia, II, III, IV, V, or any combination thereof, and an active agent.

[0313] In certain embodiments, the composition may include a drug described herein. For example, in certain embodiments, the drug is any chemical compound administered to a subject and can be delivered using the particles or nanoparticles described herein. The drug may be an organic molecule (e.g., a therapeutic agent, a drug), an inorganic molecule, a small molecule, an organometallic compound, a metal, a nucleic acid, a protein, an amino acid, a peptide, a polypeptide, a polynucleotide, a targeting agent, an isotope-labeled organic or inorganic molecule, a vaccine, an immunological agent, or an agent useful in a bioprocess. In certain embodiments, the drug is a polynucleotide. In some embodiments, the drug includes mRNA encoding at least one antigenic polypeptide or an immunogenic fragment thereof and can induce an immune response against the antigenic polypeptide.

[0314] In some embodiments, mRNA encoding at least one antigenic polypeptide or an immunogenic fragment thereof capable of inducing an immune response against the antigenic polypeptide is encapsulated by the nanoparticles.

[0315] In some aspects, disclosed herein are pharmaceutical compositions comprising a pharmaceutically acceptable carrier, nanoparticles comprising a compound of Formula I, Ia, II, III, IV, V, or any combination thereof, and mRNA encoding at least one antigenic polypeptide or immunogenic fragment thereof capable of inducing an immune response against the antigenic polypeptide.

[0316] nanoparticles Described herein are compositions comprising 20% to 80% of a compound described herein (e.g., a compound of Formula I, Ia, II, III, IV, V, or any combination thereof), greater than 0% to 5% of a polyethylene glycol lipid, greater than 0% to 40% of a helper lipid, 0% to 80% of a sterol, and

[0317] Lipid nanoparticles containing an active agent encapsulated in the nanoparticles.

[0318] Described herein are compositions comprising an effective amount of the lipid nanoparticles described herein and a pharmaceutically acceptable carrier.

[0319] In one aspect, the present disclosure provides nanoparticles, the nanoparticles comprising a compound of A, I, Ia, II, III, IV, V, or any combination thereof, and optionally a helper lipid, a polyethylene glycol lipid, a sterol, or any combination thereof, and

[0320] It comprises an active agent encapsulated in nanoparticles.

[0321] In one aspect, the present disclosure provides nanoparticles, the nanoparticles comprising a compound of A, I, Ia, II, III, IV, V, or any combination thereof, and a helper lipid, a polyethylene glycol lipid, and / or a sterol, and

[0322] It comprises an active agent encapsulated in nanoparticles.

[0323] In one aspect, the present disclosure provides a nanoparticle, the nanoparticle comprising a compound of A, I, Ia, II, III, IV, V, or any combination thereof, a helper lipid, a polyethylene glycol lipid, and a sterol, and

[0324] It comprises an active agent encapsulated in nanoparticles.

[0325] In one aspect, the present disclosure provides nanoparticles, the nanoparticles comprising a compound of formula Y and a helper lipid, a polyethylene glycol lipid, and / or a sterol, and

[0326] It comprises an active agent encapsulated in nanoparticles.

[0327] In one aspect, the present disclosure provides nanoparticles, the nanoparticles comprising a compound of Formula I and a helper lipid, a polyethylene glycol lipid, and / or a sterol, and

[0328] It comprises an active agent encapsulated in nanoparticles.

[0329] In one aspect, the present disclosure provides nanoparticles, the nanoparticles comprising a compound of formula Ia and a helper lipid, a polyethylene glycol lipid, and / or a sterol, and

[0330] It comprises an active agent encapsulated in nanoparticles.

[0331] In one aspect, the present disclosure provides nanoparticles, the nanoparticles comprising a compound of Formula II and a helper lipid, a polyethylene glycol lipid, and / or a sterol, and

[0332] It comprises an active agent encapsulated in nanoparticles.

[0333] In one aspect, the present disclosure provides nanoparticles, the nanoparticles comprising a compound of formula III and a helper lipid, a polyethylene glycol lipid, and / or a sterol, and

[0334] It comprises an active agent encapsulated in nanoparticles.

[0335] In one aspect, the present disclosure provides nanoparticles, the nanoparticles comprising a compound of formula IV and a helper lipid, a polyethylene glycol lipid, and / or a sterol, and

[0336] It comprises an active agent encapsulated in nanoparticles.

[0337] In one aspect, the present disclosure provides nanoparticles, the nanoparticles comprising a compound of formula V and a helper lipid, a polyethylene glycol lipid, and / or a sterol, and

[0338] It comprises an active agent encapsulated in nanoparticles.

[0339] Various compounds of formula Y, I, Ia, II, III, IV, or V are described above in the "Compounds" section. In some embodiments, the nanoparticles comprise a compound of formula Y, I, Ia, II, III, IV, V, or any combination thereof; The active agent encapsulated in the nanoparticles is present in a molar ratio of 20% to 80%.

[0340] In some embodiments, the nanoparticles comprise a helper lipid. In some embodiments, the helper lipid can be a non-cationic lipid. In some embodiments, the non-cationic lipid can be 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1-stearoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (SOPE), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphotidylcholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphoethanoic acid, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphoethanoic acid, ... The non-cationic lipid may include, but is not limited to, 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine (DPPE), 1,2-dimyristoyl-sn-glycero-3-phosphorylcholine (DMPC), 1,2-dioleoyl-5 / 7-glycero-3-phospho-(l'-rac-glycerol) (DOPG), 1-palmitoyl-2-linoleoyl-sn-glycero-3-phosphocholine (PLPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1-stearoyl-2-myristoyl-sn-glycero-3-phosphocholine (SMPC), or a combination thereof. In one embodiment, the non-cationic lipid is 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE). In one embodiment, the non-cationic lipid is 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE). In some embodiments, the non-cationic lipid is 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC). In some embodiments, the non-cationic lipid is 1-stearoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (SOPE). Although some non-cationic lipids are described herein, additional non-cationic lipids can also be used in combination with the compounds disclosed herein.

[0341] In some embodiments, the helper lipid may be present at a molar ratio of at least 0% (e.g., at least 5%, at least 10%, at least 20%, at least 30%, or at least 40%). In some embodiments, the helper lipid may be present at a molar ratio of 40% or less (e.g., 30% or less, 20% or less, 10% or less, 5% or less, 1% or less, or 0.5% or less).

[0342] The helper lipid may be present in a molar ratio ranging from any of the minimum values described above to any of the maximum values described above. For example, in some embodiments, the helper lipid may be present in a molar ratio of 0% to 40% (e.g., greater than 0% to 30%, greater than 0% to 20%, greater than 0% to 10%, greater than 0% to 5%, greater than 0% to 1%, greater than 0% to 0.5%, 1% to 30%, 1% to 20%, 1% to 10%, 1% to 5%, 5% to 30%, 5% to 20%, 5% to 10%, 10% to 30%, 10% to 20%, 20% to 30%, 20% to 40%, or 30% to 40%).

[0343] In some embodiments, the nanoparticles comprise polyethylene glycol lipids (PEG lipids). PEG lipids are incorporated to form a hydrophilic outer layer, stabilizing the particles. Non-limiting examples of polyethylene glycol lipids include PEG-modified lipids such as PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, and PEG-modified dialkylglycerol. Representative polyethylene glycol lipids include DMG-PEG, DLPE-PEG, DMPE-PEG, DPPC-PEG, and DSPE-PEG. In one embodiment, the polyethylene glycol lipid is 1,2-dimyristoyl-sn-glycerol, methoxypolyethylene glycol (DMG-PEG). In one embodiment, the polyethylene glycol lipid is 1,2-dimyristoyl-sn-glycerol, methoxypolyethylene glycol-2000 (DMG-PEG2000). DMG-PEGXXXX means 1,2-dimyristoyl-sn-glycerol, methoxypolyethylene glycol-XXXX, where XXXX represents the molecular weight of the polyethylene glycol moiety (e.g., DMG-PEG2000 or DMG-PEG5000).

[0344] In some embodiments, the polyethylene glycol lipid may be present in a molar ratio of at least 0% (e.g., at least 0.25%, at least 0.5%, at least 0.75%, at least 1%, at least 1.5%, at least 2%, at least 3%, at least 4%, or at least 5%). In some embodiments, the polyethylene glycol lipid may be present in a molar ratio of 5% or less (e.g., 4% or less, 3% or less, 2% or less, 1% or less, or 0.5% or less).

[0345] The polyethylene glycol lipid may be present in a molar ratio ranging from any of the minimum values described above to any of the maximum values described above. For example, in some embodiments, the polyethylene glycol lipid may be present in a molar ratio of 0% to 5% (e.g., greater than 0% to 4%, greater than 0% to 3%, greater than 0% to 2%, greater than 0% to 1%, greater than 0% to 0.5%, 1% to 5%, 1% to 4%, 1% to 3%, 1% to 2%, 2% to 5%, 2% to 4%, 2% to 3%, 3% to 5%, 3% to 4%, or 4% to 5%). In one embodiment, the molar ratio of the polyethylene glycol lipid is 0.75%.

[0346] In some embodiments, nanoparticles comprise sterol.Sterol is well known to those skilled in the art and generally refers to a compound that has a perhydrocyclopentanophenanthrene ring system and has one or more OH substituents.Examples of sterol include but are not limited to cholesterol, campesterol, ergosterol, sitosterol, etc.

[0347] In some embodiments, the sterol is selected from cholesterol-based lipids. In some embodiments, the one or more cholesterol-based lipids are selected from cholesterol, PEGylated cholesterol, DC-Chol (N,N-dimethyl-N-ethylcarboxamidocholesterol), 1,4-bis(3-N-oleylaminopropyl)piperazine, or a combination thereof.

[0348] Sterols can be used to adjust the permeability and fluidity of particles based on their function in cell membranes, hi one embodiment, the sterol is cholesterol.

[0349] In some embodiments, the molar ratio of sterols is at least 0% (e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, or at least 80%). In some embodiments, the molar ratio of sterols is 80% or less (e.g., 70% or less, 60% or less, 50% or less, 40% or less, 30% or less, 20% or less, 10% or less, 5% or less, or 1% or less).

[0350] The sterol may be present in a molar ratio ranging from any of the minimum values above to any of the maximum values above. In some embodiments, the sterol is present in a molar ratio of 0% to 80% (e.g., 0% to 70%, 0% to 60%, greater than 0% to 50%, greater than 0% to 40%, greater than 0% to 30%, greater than 0% to 20%, greater than 0% to 10%, greater than 0% to 5%, greater than 0% to 1%, 5% to 70%, 5% to 60%, 5% to 50%, 5% to 40%, 5% to 30%, 5% to 20%, 5% to 10%, 10% to 70%, 10% to 60%, 10% to 50%, 10% to 40%, 10% to 30%, 10% to 20%, or 20%. The sterols may be present in a molar ratio of 0%, 20% to 70%, 20% to 60%, 20% to 50%, 20% to 40%, 20% to 30%, 30% to 70%, 30% to 60%, 30% to 50%, 30% to 40%, 40% to 70%, 40% to 60%, 40% to 50%, 50% to 70%, 50% to 60%, 60% to 70%, 60% to 80%, 70% to 80%, 10% to 80%, 20% to 80%, 30% to 80%, 40% to 80%, or 50% to 80%. In some embodiments, the nanoparticles comprise sterols at a molar ratio of 40%.

[0351] In some embodiments, the nanoparticles may comprise 40%-60% of a compound described herein, 1%-2% polyethylene glycol lipid, 8%-12% helper lipid, 35%-40% sterol, and an active agent encapsulated in the nanoparticles.

[0352] In one embodiment, described are nanoparticles comprising 50% of a compound described herein, 1.5% polyethylene glycol lipid, 10% helper lipid (e.g., DSPC), 38.5% sterol (e.g., cholesterol), and an active agent encapsulated in the nanoparticles.

[0353] In one embodiment, the present disclosure provides a nanoparticle, the nanoparticle comprising:

[0354] A compound of formula Y, I, Ia, II, III, IV, V, or any combination thereof; and

[0355] It comprises an active agent encapsulated in nanoparticles.

[0356] In some embodiments, described are nanoparticles, the nanoparticles comprising:

[0357] A compound of formula Y, I, Ia, II, III, IV, V, or any combination thereof;

[0358] 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE),

[0359] 1,2-Dimyristoyl-sn-glycerol, methoxypolyethylene glycol (DMG-PEG 2000 ),

[0360] Cholesterol, and

[0361] It comprises an active agent encapsulated in nanoparticles.

[0362] In one embodiment, the present disclosure provides a nanoparticle, the nanoparticle comprising:

[0363] A compound of formula Y, I, Ia, II, III, IV, V, or any combination thereof;

[0364] 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE),

[0365] 1,2-Dimyristoyl-sn-glycerol, methoxypolyethylene glycol (DMG-PEG 2000 ),

[0366] Cholesterol, and

[0367] It comprises an active agent encapsulated in nanoparticles.

[0368] In one embodiment, the present disclosure provides a nanoparticle, the nanoparticle comprising:

[0369] A compound of formula Y, I, Ia, II, III, IV, V, or any combination thereof;

[0370] 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC),

[0371] 1,2-Dimyristoyl-sn-glycerol, methoxypolyethylene glycol (DMG-PEG 2000 ), and

[0372] Contains cholesterol.

[0373] In one embodiment, the present disclosure provides a nanoparticle, the nanoparticle comprising:

[0374] A compound of formula Y, I, Ia, II, III, IV, V, or any combination thereof;

[0375] 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE),

[0376] 1,2-Distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (DSPE-PEG 2000 ),

[0377] Cholesterol, and

[0378] It comprises an active agent encapsulated in nanoparticles.

[0379] In one embodiment, the nanoparticles may further comprise an active agent. In one embodiment, the nanoparticles may further comprise a therapeutic agent. In one embodiment, the nanoparticles may further comprise a diagnostic agent. In one embodiment, the nanoparticles may further comprise a prophylactic agent.

[0380] activator As used herein, "active agent" refers to a therapeutic, diagnostic, or prophylactic agent. As discussed herein, therapeutic agents can be released from the disclosed compounds, compositions, and systems in a biologically active form.

[0381] Furthermore, as used herein, the term "therapeutic agent" is understood to refer to one or more therapeutic agents, active ingredients, or substances that can be used to treat a medical condition. Therapeutic agents include synthetic or natural bioactive compounds or compositions of substances that, when administered to an organism (human or non-human animal), induce a desired pharmacological, immunogenic, and / or physiological effect through local and / or systemic action. Thus, the term encompasses those compounds or chemicals traditionally considered to be drugs, vaccines, or biopharmaceuticals, including molecules such as proteins, peptides, hormones, nucleic acids, and gene constructs. Examples of therapeutic agents are found in well-known sources such as the Merck Index (14th ed.), Physicians' Desk Reference (64th ed.), and The Pharmacological Basis of Therapeutics (12th ed.), and include, but are not limited to, pharmaceuticals, essential amino acids, vitamins and minerals such as calcium, iron, potassium, zinc, vitamin B12, and the like, substances used in the treatment, prevention, diagnosis, cure, or mitigation of disease or disorder, substances that affect the structure or function of the body, or prodrugs that become biologically active or more active after being placed in a physiological environment. For example, the term "therapeutic agent" includes adjuvants, antibacterial agents (including antibiotics, antivirals, antiparasitics, and antifungals), anti-inflammatory agents (including steroids and nonsteroidal anti-inflammatory agents), anticoagulants, eye drops, gastrointestinal agents, antiplatelet agents, antiseptics, steroids, antitumor agents, anticancer agents, antigens, antibodies (e.g., cetuximab, anti-CD24 antibodies, panitumumab, bevacizumab), contraceptives, progesterone agents, anticholinergic agents, nutritional agents, analgesics, and combinations of analgesics such as acetaminophen and acetylsalicylic acid. combinations, anesthetics such as lidocaine and xylocaine, appetite suppressants such as dexadrine and phendimetrazine tartrate, antiepileptics, local anesthetics and general anesthetics, hypnotics, sedatives, antipsychotics, neuroleptics, antidepressants such as isocarboxazid and amoxapine, antianxiety drugs, antagonists, neuron blockers, anticholinergics and cholinergic agents, antimuscarinic and muscarinic agents, antiparkinsonian drugs, antialzheimer's drugs, antiadrenergic drugs, antiarrhythmic drugs, antihypertensive drugs, insulin, progestins,Hormones and nutrients such as estrogen, corticoids, glucocorticoids, and androgens, anti-arthritis drugs such as methylprednisolone and ibuprofen, anti-asthma drugs such as terbutaline sulfate, theophylline, and ephedrine, anticonvulsants such as phenytoin sodium and diazepam, antiallergic drugs, antihistamines such as diphenhydramine HCl and chlorpheniramine maleate, antiemetics, antitumor agents, antipruritics, antipyretics, belladonna alkaloids, antispasmodics such as dicyclomine hydrochloride, prazosin HCl, nitroglycerin, propranolol Cardiovascular drugs such as benzodiazepine HCl, hydralazine HCl, pancrelipase, succinate dehydrogenase, vasoactive drugs, cardiovascular preparations (including calcium channel blockers, beta-blockers, beta-agonists, and antiarrhythmic drugs), antihypertensive drugs, diuretics such as furosemide and spironolactone, vasodilators, central nervous system stimulants, cough and cold medicines, decongestants, diagnostic drugs, bone growth stimulants and bone resorption inhibitors, muscle relaxants, psychostimulants, sedatives, tranquilizers such as thorazine, diazepam, chlorpromazine HCl, reserpine, chlordiazepoxide HCl, rantidine HCl, cimetidine HCl anti-ulcer drugs such as anti-asthmatic drugs, anti-diarrheal drugs, anti-obesity drugs, anti-thrombotic drugs, antitussive drugs, anti-uric acid drugs, anti-anginal drugs, appetite suppressants, expectorants, hyperglycemic drugs, hypoglycemic drugs, thyroid and anti-thyroid drugs, tissue growth drugs, uterine relaxants, immunomodulatory agents including, for example, cytokines, interleukins, interferons, colony stimulating factors, tumor necrosis factors, etc., immunosuppressants such as rapamycin, tacrolimus, immunological agents, antigens, factors, growth factors, amino acids, peptides and proteins, and fragments thereof, whether naturally occurring, chemically synthesized or recombinantly produced. any of the above), for example, LHRH, somatostatin, calcitonin, growth hormone, glucagon-like peptide, growth releasing factor, angiotensin, FSH, EGF, bone morphogenetic protein (BMP), erythropoietin (EPO), interferon, interleukin, collagen, fibrinogen, insulin, factor VIII, factor IX, Enbrel (registered trademark), Rituxam (registered trademark), Herceptin (registered trademark), alpha glucosidase, Serazyme / Ceredose (registered trademark), vasopressin, ACTH, human serum albumin, gamma globulin,These include compounds or compositions used in all major therapeutic areas, including, but not limited to, structural proteins, blood product proteins, complex proteins, antigens or antigenic polypeptides, enzymes, antibodies, monoclonal antibodies, etc., nucleic acid molecules (polymeric forms of two or more nucleotides, including polynucleotides, either ribonucleotides (RNA) or deoxyribonucleotides (DNA), both double-stranded and single-stranded molecules, gene constructs, expression vectors, antisense molecules, etc.), small molecules (e.g., doxorubicin), and other biologically active macromolecules such as proteins and enzymes. The agent can be a bioactive agent used in medical applications, including veterinary applications, and in agriculture, such as in plants, and other fields. In certain embodiments of the present disclosure, the agent to be delivered can be a mixture of active agents.

[0382] Representative examples of antibiotics include amikacin, amoxicillin, ampicillin, atovaquone, azithromycin, aztreonam, bacitracin, carbenicillin, cefadroxil, cefazolin, cefdinir, cefditoren, cefepime, cefiderocol, cefoperazone, cefotetan, cefoxitin, cefotaxime, cefpodoxime, cefprozil, ceftaroline, ceftazidime, ceftibuten, and ceftizolinone. oxime, ceftriaxone, chloramphenicol, colistimethimesate, cefuroxime, cephalexin, cephradine, cilastatin, cinoxacin, ciprofloxacin, clarithromycin, clindamycin, dalbavancin, dalfopristin, daptomycin, demeclocycline, dicloxacillin, doripenem, doxycycline, eravacycline, ertapenem, erythromycin, fidaxoma Isin, fosfomycin, gatifloxacin, gemifloxacin, gentamicin, imipenem, lefamulin, lincomycin, linezolid, lomefloxacin, loracarbef, meropenem, metronidazole, minocycline, moxifloxacin, nafcillin, nalidixic acid, neomycin, norfloxacin, ofloxacin, omadacycline, oritavancin, oxacillin, oxytetracycline , paromomycin, penicillin, pentamidine, piperacillin, plazomycin, quinupristin, rifaximin, sarecycline, secnidazole, sparfloxacin, spectinomycin, sulfamethoxazole, sulfisoxazole, tedizolid, telavancin, telitromycin, ticarcillin, tigecycline, tobramycin, trimethoprim, trovafloxacin, and vancomycin.

[0383] Representative examples of antiviral agents include abacavir, acyclovir, adefovir, amantadine, amprenavir, atazanavir, baravir, baloxavir marboxil, boceprevir, cidofovir, cobicistat, daclatasvir, darunavir, delavirdine, didanosine, docasanol, dolutegravir, doravirine, ecoleaver, edoxudine, efavirenz, elvitegravir, emtricitabine, enfuvirtide, entecavir, etravirine, famciclovir, fomivirsen, fosamprenavir, forscarnet, fosnonet, famciclovir, favipravir, fomivirsen, foscavir, ganciclovir, ibacitabine, idoxuridine, indinavir, inosine, inosine pranobex, interferon type I, interferon type II, interferon type III, lamivudine, letermovir fluticasone, letermovir, lopinavir, loviride, maraviroc, methisazone, moroxydine, nelfinavir, nevirapine, nitazoxanide, oseltamivir, peginterferon alfa-2a, peginterferon alfa-2b, penciclovir, peramivir, pleconaril, podophyllotoxin, pyramidine, raltegravir, remdesevir, ribavirin, rilpivirine, rimantadine, lintatolimod, mogamulizumab, fluticasone, fluoxetine, fluoxetine-10 ... These include, but are not limited to, lunupiravir, ritonavir, saquinavir, simeprevir, sofosbuvir, stavudine, talabivirine, telaprevir, telbivudine, tenofovir alafenamide, tenofovir disoproxil, tenofovir, tipranavir, trifluridine, trizivir, tromantadine, umifenovir, valacyclovir, valganciclovir, vidarabine, zalcitabine, zanamivir, and zidovudine.

[0384] Representative examples of anticoagulants include, but are not limited to, heparin, warfarin, rivaroxaban, dabigatran, apixaban, edoxaban, enoxaparin, and fondaparinux.

[0385] Representative examples of antiplatelet agents include, but are not limited to, clopidogrel, ticagrelor, prasugrel, dipyridamole, dipyridamole / aspirin, ticlopidine, and eptifibatide.

[0386] Representative examples of antifungal agents include, but are not limited to, voriconazole, itraconazole, posaconazole, fluconazole, ketoconazole, clotrimazole, isavuconazonium, miconazole, caspofungin, anidulafungin, micafungin, griseofulvin, terbinafine, flucytosine, terbinafine, nystatin, and amphotericin B.

[0387] Representative examples of steroidal anti-inflammatory drugs include, but are not limited to, hydrocortisone, dexamethasone, prednisolone, prednisone, triamcinolone, methylprednisolone, budesonide, betamethasone, cortisone, and deflazacort. Representative examples of non-steroidal anti-inflammatory drugs include, but are not limited to, ibuprofen, naproxen, ketoprofen, tolmetin, etodolac, fenoprofen, flurbiprofen, diclofenac, piroxicam, indomethacin, sulindax, meloxicam, nabumetone, oxaprozin, mefenamic acid, and diflunisal.

[0388] Examples of other active agents include chloroquine, hydrochloroquine, pyridoxal phosphate, vitamin D, and vitamin C.

[0389] Representative examples of anti-cytokine or immunomodulatory agents include, but are not limited to, tocilizumab, sarilumab, bevacizumab, fingolimod, imiquimod, and eculizumab.

[0390] Immunotherapeutics include, but are not limited to, anti-CD40 antibodies, anti-PDL1 antibodies (e.g., atezolizumab, durvalumab, avelumab), anti-PD1 antibodies, anti-CTLA4 antibodies, programmed cell death protein 1 (PD-1) inhibitors, or programmed cell death protein ligand 1 or 2 inhibitors, including, for example, nivolumab (BMS), pembrolizumab (Merck), pidilizumab (CureTech / Teva), AMP-244 (Amplimmune / GSK), BMS-936559 (BMS), and MEDI4736 (Roche / Genentech), or combinations thereof.

[0391] Representative examples of contraceptives include, but are not limited to, progestins, estrogens, or combinations thereof. For example, suitable progestins include, but are not limited to, natural and synthetic compounds with progestational activity, such as progesterone, chlormadinone acetate, norethindrone, cyproterone acetate, norethindrone acetate, desogestrel, levonorgestrel, drospirenone, trimegestone, norgestrel, norgestimate, norelgestromin, etonogestrel, gestodene, and other natural and / or synthetic gestagens. For example, suitable estrogens include, but are not limited to, natural and synthetic compounds with estrogenic activity, such as estradiol (17β-estradiol), 17α-estradiol, estriol, estrone, and their esters, such as acetate, sulfate, valerate, or benzoate esters of these compounds (e.g., estradiol 17β-cypionate, estradiol 17-propionate, estradiol 3-benzoate, and piperazine estrone sulfate), ethinyl estradiol, conjugated estrogens (natural and synthetic), mestranol, agonist antiestrogens, and selective estrogen receptor modulators. Other examples of contraceptives include gonodotropin-releasing hormone (GnRh) or analogs thereof, such as deslorelin, avorelin, leuprolide, triptorelin, nafarelin, goserelin, buserelin, and fertirelin.

[0392] The term "steroid" refers to compounds belonging to or related to the following exemplary families of compounds: corticosteroids, mineral steroids, and sex steroids (e.g., including potentially androgenic or estrogenic, or antiandrogenic and antiestrogenic molecules). These include, for example, prednisone, prednisolone, methylprednisolone, triamcinolone, fluocinolone, aldosterone, spironolactone, danazol (also known as Optina), and others. In some embodiments, the therapeutic agent can include a steroid.

[0393] Exemplary cancer therapeutic or anti-cancer agents include, but are not limited to, antimetabolite anti-cancer agents, antimitotic anti-cancer agents, and combinations thereof. A variety of antimetabolite and anti-mitotic anti-cancer agents can be used in the methods and compositions described herein, including single antimetabolite and anti-mitotic anti-cancer agents or combinations of multiple antimetabolites.

[0394] Antimetabolic anticancer drugs are typically structurally similar to natural metabolites involved in normal metabolic processes in cancer cells, such as nucleic acid and protein synthesis. However, antimetabolites are significantly different from natural metabolites and therefore interfere with cancer cell metabolic processes. Within the cell, antimetabolites are mistaken for similar metabolites and are processed by the cell in the same way as normal compounds. The presence of the "decoy" metabolites prevents the cell from performing vital functions, preventing cell growth and survival. For example, antimetabolites can exert cytotoxicity by substituting these incorrect nucleotides into the cell's DNA, preventing cell division, or by inhibiting key cellular enzymes, preventing DNA replication.

[0395] In one embodiment, therefore, the anti-metabolite anti-cancer agent is a nucleotide or nucleotide analog. In certain embodiments, for example, the anti-metabolite can include a purine (e.g., guanine or adenosine) or analog thereof, or a pyrimidine (cytidine or thymidine) or analog thereof, with or without an attached sugar moiety.

[0396] Anti-metabolite anti-cancer agents suitable for use in the present disclosure can generally be classified according to the metabolic process they affect and can include, but are not limited to, analogs and derivatives of folic acid, pyrimidines, purines, and cytidine. Thus, in one aspect, the anti-metabolite(s) is selected from the group consisting of cytidine analogs, folic acid analogs, purine analogs, pyrimidine analogs, and combinations thereof.

[0397] In one particular embodiment, for example, the antimetabolite is a cytidine analog. According to this embodiment, for example, the cytidine analog can be selected from the group consisting of cytarabine (cytosine arabinoside), azacytidine (5-azacytidine), and salts, analogs, and derivatives thereof. In another specific embodiment, for example, the antimetabolite is a folic acid analog. Folic acid analogs or antifolates generally act by inhibiting dihydrofolate reductase (DHFR), an enzyme involved in the formation of nucleotides; when this enzyme is inhibited, nucleotides are not formed, preventing DNA replication and cell division. In certain embodiments, for example, the folic acid analog can be selected from the group consisting of denopterin, methotrexate (amethopterin), pemetrexed, pteropterin, raltitrexed, trimetrexate, and salts, analogs, and derivatives thereof.

[0398] In another specific embodiment, for example, the antimetabolite is a purine analog. Purine antimetabolites act by inhibiting DNA synthesis, such as by preventing the production of purines containing the nucleotides adenine and guanine, which stop DNA synthesis and cell division. Purine analogs can also be incorporated into the DNA molecule itself during DNA synthesis, preventing cell division. According to certain embodiments, for example, the purine analog may be selected from the group consisting of acyclovir, allopurinol, 2-aminoadenosine, arabinosyladenine (ara-A), azacitidine, azathiprine, 8-azaadenosine, 8-fluoroadenosine, 8-methoxyadenosine, 8-oxoadenosine, cladribine, deoxycoformycin, fludarabine, gancylovir, 8-azaguanosine, 8-fluoroguanosine, 8-methoxyguanosine, 8-oxoguanosine, guanosine diphosphate, guanosine diphosphate-β-L-2-aminofucose, guanosine diphosphate-D-arabinose, guanosine diphosphate-2-fluorofucose, guanosine diphosphate fucose, mercaptopurine (6-MP), pentostatin, thiamiprine, thioguanine (6-TG), and salts, analogs, and derivatives thereof.

[0399] In yet another particular embodiment, for example, the antimetabolite is a pyrimidine analog. Similar to the purine analogs described above, pyrimidine antimetabolites inhibit the synthesis of pyrimidine-containing nucleotides (cytosine and thymine in DNA, cytosine and uracil in RNA). Pyrimidine compounds can act as "decoys" to prevent the production of nucleotides and / or can be incorporated into growing DNA strands, leading to their termination. According to certain embodiments, for example, the pyrimidine analog is ancitabine, azacitidine, 6-azauridine, bromouracil (e.g., 5-bromouracil), capecitabine, carmofur, chlorouracil (e.g., 5-chlorouracil), cytarabine (cytosine arabinoside), cytosine, dideoxyuridine, 3'-azido-3'-deoxythymidine, 3'-dideoxycytidin-2'-ene, 3'-deoxy-3'-deoxythymidin-2'-ene, dihydrouracil, doxifluridine, enocitabine, floxuridine, 5- The pyrimidine analog is selected from the group consisting of fluorocytosine, 2-fluorodeoxycytidine, 3-fluoro-3'-deoxythymidine, fluorouracil (e.g., 5-fluorouracil (also known as 5-FU)), gemcitabine, 5-methylcytosine, 5-propynylcytosine, 5-propynylthymine, 5-propynyluracil, thymine, uracil, uridine, and salts, analogs, and derivatives thereof. In one embodiment, the pyrimidine analog is other than 5-fluorouracil. In another embodiment, the pyrimidine analog is gemcitabine or a salt thereof.

[0400] In certain embodiments, the antimetabolite is selected from the group consisting of 5-fluorouracil, capecitabine, 6-mercaptopurine, methotrexate, gemcitabine, cytarabine, fludarabine, pemetrexed, and salts, analogs, derivatives, and combinations thereof. In other embodiments, the antimetabolite is selected from the group consisting of capecitabine, 6-mercaptopurine, methotrexate, gemcitabine, cytarabine, fludarabine, pemetrexed, and salts, analogs, derivatives, and combinations thereof. In one particular embodiment, the antimetabolite is other than 5-fluorouracil. In certain preferred embodiments, the antimetabolite is gemcitabine or a salt thereof (e.g., gemcitabine HCl (Gemzar®)).

[0401] Other antimetabolite anticancer agents include, among others, acantifolic acid, aminothiadiazole, brequinar sodium, Ciba-Geigy CGP-30694, cyclopentylcytosine, cytarabine phosphate stearate, cytarabine conjugate, Lilly DATHF, Merreldow DDFC, desaguanine, dideoxycytidine, dideoxyguanosine, Didox, Yoshitomi DMDC, Wellcome EHNA, Merck EX-015, fazarabine, fludarabine phosphate, N-(2'-furanidyl)-5-fluorouracil, Daiichi Pharmaceutical FO-152, 5-FU-fibrinogen, isopropylpyrrolidine, Lilly LY-188011, Lilly LY-264618, metbenzaprim, Wellcome MZPES, norspermidine, NCI NSC-127716, NCI NSC-264880, NCI The inhibitor may be selected from the group consisting of, but is not limited to, NSC-39661, NCI NSC-612567, Warner-Lambert PALA, pentostatin, piritrexim, plicamycin, Asahi Kasei PL-AC, Takeda TAC-788, tiazofurin, Elbamont TIF, tyrosine kinase inhibitors, Taiho UFT, and ulisitin.

[0402] In one embodiment, the antimitotic anticancer agent is a microtubule inhibitor or microtubule stabilizer. Generally, microtubule stabilizers, such as taxanes and epothilones, bind to the inner surface of β-microtubule strands, promoting the nucleation and elongation steps of the polymerization reaction and enhancing microtubule assembly by lowering the critical tubulin subunit concentration required for microtubule assembly. While microtubule inhibitors, such as vinca alkaloids, prevent microtubule assembly, microtubule stabilizers, such as taxanes, shorten the lag time and dramatically shift the dynamic equilibrium between tubulin dimers and microtubule polymers toward polymerization. In one embodiment, the microtubule stabilizer is a taxane or epothilone. In another embodiment, the microtubule inhibitor is a vinca alkaloid. In some embodiments, the anticancer agent may comprise a taxane or a derivative or analog thereof. Taxanes may be naturally occurring compounds or related forms thereof with antitumor properties, or may be chemically synthesized compounds or derivatives thereof. Taxanes are a family of terpenes derived primarily from the Pacific yew tree, Taxus cuspidata, and include, but are not limited to, paclitaxel (Taxol®) and docetaxel (Taxotere®), which have activity against certain tumors, particularly breast and ovarian cancers. In one aspect, the taxane is docetaxel or paclitaxel. Paclitaxel is a preferred taxane and is considered an antimitotic agent that stabilizes microtubules by promoting the assembly of microtubules from tubulin dimers and preventing depolymerization. This stability inhibits the normal dynamic reorganization of the microtubule network, which is essential for important interphase and mitotic cellular functions.

[0403] Also included are various known taxane derivatives, including both hydrophilic and hydrophobic derivatives. Taxane derivatives include, but are not limited to, the galactose and mannose derivatives described in International Patent Application No. WO 99 / 18113, the piperazino and other derivatives described in WO 99 / 14209, the taxane derivatives described in WO 99 / 09021, WO 98 / 22451, and U.S. Patent No. 5,869,680, the 6-thio derivatives described in WO 98 / 28288, the sulfenamide derivatives described in U.S. Patent No. 5,821,263, the deoxygenated paclitaxel compounds described in U.S. Patent No. 5,440,056, and the taxol derivatives described in U.S. Patent No. 5,415,869. As mentioned above, this includes prodrugs of paclitaxel, including, but not limited to, those described in WO 98 / 58927, WO 98 / 13059, and U.S. Patent No. 5,824,701. The taxane can be a taxane conjugate, such as, for example, paclitaxel-PEG, paclitaxel-dextran, paclitaxel-xylose, docetaxel-PEG, docetaxel-dextran, docetaxel-xylose, etc. Other derivatives are described in various references, such as "Synthesis and Anticancer Activity of Taxol Derivatives," D.G.I. Kingston et al., Studies in Organic Chemistry, vol. 26, entitled "New Trends in Natural Products Chemistry" (1986), Atta-ur-Rabman, P.W. Le Quesne, Eds. (Elsevier, Amsterdam 1986). Each of these references is incorporated herein by reference in its entirety.

[0404] A variety of taxanes can be readily prepared using techniques known to those skilled in the art (see, also, WO 94 / 07882, WO 94 / 07881, WO 94 / 07880, WO 94 / 07876, WO 93 / 23555, WO 93 / 10076, U.S. Pat. Nos. 5,294,637, 5,283,253, 5,279,949, 5,274,137, 5,202,448, 5,200,534, 5,229,529, and European Patent No. 590,267), each of which is incorporated by reference in its entirety, or are available from a variety of commercial sources, including, for example, Sigma-Aldrich Co., St. Louis, Mo.

[0405] Alternatively, the antimitotic anticancer agent can be a microtubule inhibitor, and in a preferred embodiment, the microtubule inhibitor is a vinca alkaloid.Generally, vinca alkaloids are mitotic spindle poisons.Vinca alkaloids act during mitosis, when chromosomes are divided and begin to move along the mitotic spindle tubules toward one of their poles before cell separation.The action of these spindle poisons causes chromosome dispersion and spindle disruption during mitosis, affecting cell reproduction.In a specific embodiment, for example, the vinca alkaloid is selected from the group consisting of vinblastine, vincristine, vindesine, vinorelbine, and their salts, analogs, and derivatives.

[0406] The antimitotic anticancer agent can be an epothilone. Generally, members of the epothilone class of compounds stabilize microtubule function according to a mechanism similar to that of taxanes. Epothilones can also cause cell cycle arrest at the G2-M transition, leading to cytotoxicity and ultimately apoptosis. Suitable epithilones include epothilone A, epothilone B, epothilone C, epothilone D, epothilone E, and epothilone F, as well as salts, analogs, and derivatives thereof. A particular epothilone analog is ixabepilone (Ixempra™), an epothilone B analog.

[0407] In certain embodiments, the antimitotic anticancer agent is selected from the group consisting of taxanes, epothilones, vinca alkaloids, and salts and combinations thereof. Thus, for example, in one embodiment, the antimitotic agent is a taxane. More preferably, in this embodiment, the antimitotic agent is paclitaxel or docetaxel, and even more preferably paclitaxel. In another embodiment, the antimitotic agent is an epothilone (e.g., an epothilone B analog). In another embodiment, the antimitotic agent is a vinca alkaloid.

[0408] Examples of anti-cancer agents that can be used in the present disclosure include, but are not limited to, platinum complexes such as thalidomide, cisplatin (cis-DDP), oxaliplatin, and carboplatin; anthracenediones such as mitoxantrone; substituted ureas such as hydroxyurea; methylhydrazine derivatives such as procarbazine (N-methylhydrazine, MIH); adrenocortical suppressants such as mitotane (o,p'-DDD) and aminoglutethimide; RXR agonists such as bexarotene; and tyrosine kinase inhibitors such as sunitimibe and imatinib.

[0409] Examples of other anticancer drugs include alkylating agents, antimetabolites, natural products, hormones and antagonists, and other drugs. Alternative names are provided in parentheses. Examples of alkylating agents include nitrogen mustards such as mechlorethamine, cyclophosphanide, ifosfamide, melphalan (sarcolysin), chlorambucil, ethyleneimines and methylmelamines such as hexamethylmelamine and thiotepa, alkylsulfonates such as busulfan, nitrosoureas such as carmustine (BCNU), semustine (methyl-CCNU), lomustine (CCNU), and streptozotocin, DNA synthesis antagonists such as estramustine phosphate, and triazines such as dacarbazine (DTIC, dimethyl-triazenoimidazole carboxamide) and temozolomide. Examples of antimetabolites include folic acid analogs such as methotrexate (amethopterin), pyrimidine analogs such as fluorouracin (5-fluorouracil, 5-FU, SFU), floxuridine (fluorodeoxyuridine, FUdR), cytarabine (cytosine arabinoside), gemcitabine, purine analogs such as mercaptopurine (6-mercaptopurine, 6-MP), thioguanine (6-thioguanine, TG), pentostatin (2'-deoxycoformycin, deoxycoformycin), cladribine, fludarabine, and topoisomerase inhibitors such as amsacrine. Examples of natural products include vinca alkaloids such as vinblastine (VLB) and vincristine; taxanes such as paclitaxel, protein-bound paclitaxel (Abraxane), and docetaxel (Taxotere); epipodophyllotoxins such as etoposide and teniposide; camptothecins such as topotecan and irinotecan; antibiotics such as dactinomycin (actinomycin D), daunorubicin (daunomycin, rubidomycin), doxorubicin, histrilin, bleomycin, mitomycin (mitomycin C), idarubicin, and epirubicin; enzymes such as L-asparaginase; and biological response modifiers such as interferon alpha and interleukin 2.Examples of hormones and antagonists include luteinizing hormone-releasing hormone agonists such as buserelin, corticosteroids and related preparations such as prednisone, progestins such as hydroxyprogesterone caproate, medroxyprogesterone acetate, megestrol acetate, and the like, estrogens and related preparations such as diethylstilbestrol, ethinyl estradiol, estrogen antagonists such as tamoxifen and anastrozole, androgens and related preparations such as testosterone propionate and fluoxymesterone, androgen antagonists such as flutamide and bicalutamide, and gonadotropin-releasing hormone analogs such as leuprolide. Alternative names and trade names of these and other examples, as well as methods of use, including dosages and administration regimens, will be known to those skilled in the art.

[0410] In some embodiments, the anti-cancer agent may comprise a chemotherapeutic agent. Suitable chemotherapeutic agents include, but are not limited to, alkylating agents, antibiotics, metabolic inhibitors, hormones, plant-derived drugs and their synthetic derivatives, anti-angiogenic agents, differentiation inducers, cell growth arrest inducers, apoptosis inducers, cytotoxic agents, agents that affect cellular energetics, i.e., agents that affect cellular ATP levels and molecules / activities that regulate these levels, biological agents, e.g., monoclonal antibodies, kinase inhibitors, inhibitors of growth factors and their receptors, gene therapy agents, cell therapy, e.g., stem cells, or combinations thereof.

[0411] According to these embodiments, the chemotherapeutic agent is selected from the group consisting of cyclophosphamide, chlorambucil, melphalan, mechlorethamine, ifosfamide, busulfan, lomustine, streptozocin, temozolomide, dacarbazine, cisplatin, carboplatin, oxaliplatin, procarbazine, uramustine, methotrexate, pemetrexed, fludarabine, cytarabine, fluorouracil, floxuridine, gemcitabine, capecitabine, vinblastine, vincristine, vinorelbine, etoposide, paclitaxel, docetaxel, doxorubicin, daunorubicin, epirubicin, idarubicin, mitoxantrone, bleomycin, mitomycin, hydroxyurea, topotecan, irinotecan, amsacrine, teniposide, erlotinib hydrochloride, and combinations thereof. Each possibility represents a separate embodiment of the present invention.

[0412] Antitumor agents include abiraterone acetate, avitrexate (methotrexate), Abraxane (paclitaxel albumin-stabilized nanoparticles), ABVD, ABVE, ABVE-PC, AC, AC-T, Adcetris (brentuximab vedotin), ADE, adotrastuzumab emtansine, Adriamycin (doxorubicin hydrochloride), Adrsil (fluorouracil), afatinib dimaleate, Afinitor (everolimus), Aquinzeo (netupitant and palonosetron hydrochloride), Aldara (imiquimod), aldesleukin, and alemtuzumab. Mab, Alimta (pemetrexed disodium), Aloxi (palonosetron hydrochloride), Ambochlorin (chlorambucil), Aminolevulinic acid, Anastrozole, Aprepitant, Aredia (pamidronate disodium), Arimidex (anastrozole), Aromasin (exemestane), Alanon (nelarabine), Arsenic trioxide, Alzera (ofatumumab), Asparaginase Erwinia chrysanthemi, Avastin (bevacizumab), Axitinib, Azacitidine, BEACOPP, Besenam (Callum) bendamustine), Beleodac (belinostat), belinostat, bendamustine hydrochloride, BEP, bevacizumab, bexarotene, Bexar (tositumomab and iodine I131 tositumomab), bicalutamide, BiCNU (carmustine), bleomycin, blinatumomab, blinatumomab (blinatumomab), bortezomib, Bosurif (bosutinib), bosutinib, brentuximab vedotin, busulfan, Busulfex (busulfan), cabazitaxel, cabozantinib-S-malate, CAF, bleomycin, blinatumomab, blinatumomab Mab (blinatumomab), bortezomib, Bosurif (bosutinib), bosutinib, brentuximab vedotin, busulfan, Busulfex (busulfan), cabazitaxel, cabozantinib-S-malate, CAF, Carmbris (carmustine), carmustine, carmustine implant, Casodex (bicalutamide), CeeNU (lomustine), ceritinib, Cerbidine (daunorubicin hydrochloride), Cervarix (recombinant HPV bivalent vaccine), cetuximab, chlorambucil, chlorambucil prednisone, CHOP, cisplatin,Clafen (cyclophosphamide), clofarabine, Clofarex (clofarabine), Chlor (clofarabine), CMF, Cometriq (cabozantinib-S-malate), COPP, COPP-ABV, Cosmegen (dactinomycin), crizotinib, CVP, cyclophosphamide, Cyphos (ifosfamide), Cyramza (ramucirumab), cytarabine, cytarabine, liposomal, Cytosar-U (cytarabine), Cytoxan (cyclophosphamide), dabrafenib, dacarbazine, Dacogen (decitabine), dactinomycin , dasatinib, daunorubicin hydrochloride, decitabine, degarelix, denileukin diftitox, denosumab, DepoCyte (liposomal cytarabine), DepoForm (liposomal cytarabine), dexrazoxane hydrochloride, dinutuximab, docetaxel, Doxil (doxorubicin hydrochloride liposomal), doxorubicin hydrochloride, doxorubicin hydrochloride liposomal, Dox-SL (doxorubicin hydrochloride liposomal), DTIC-Dome (dacarbazine), Efudex (fluorouracil), ERYTECH (rasburicase), Elence (epirubicin hydrochloride) phosphate), Eloxatin (oxaliplatin), eltrombopag olamine, Emend (aprepitant), enzalutamide, epirubicin hydrochloride, EPOCH, Erbitux (cetuximab), eribulin mesylate, Erivedge (vismodegib), erlotinib hydrochloride, Erwinaze (asparaginase Erwinia chrysanthemi), Etopophos (etoposide phosphate), etoposide, etoposide phosphate, Everset (doxorubicin hydrochloride liposomal), everolimus, Evista (raloxifene hydrochloride), exemestane, Fareston (treonam) Miphen), Faridak (panobinostat), Faslodex (fulvestrant), FEC, Femara (letrozole), filgrastim, Fludara (fludarabine phosphate), fludarabine phosphate, Fluoroplex (fluorouracil), fluorouracil, Folex (methotrexate), Folex PFS (methotrexate), FOLFIRI, FOLFIRI-bevacizumab, FOLFIRI-cetuximab, FOLFIRINOX, FOLFOX, Folotin (pralatrexate), FU-LV, fulvestrant,Gardasil (recombinant HPV quadrivalent vaccine), Gardasil 9 (recombinant HPV 9-valent vaccine), Gazyva (obinutuzumab), gefitinib, gemcitabine hydrochloride, gemcitabine cisplatin, gemcitabine oxaliplatin, gemtuzumab ozogamicin, Gemzar (gemcitabine hydrochloride), Jilotrif (afatinib dimaleate), Glivec (imatinib mesylate), Gliadel (carmustine implant), Gliadel wafer (carmustine implant), glucarpidase, goserelin acetate, Halaven (eribulin mesylate) ), Herceptin (trastuzumab), HPV bivalent vaccine (recombinant), HPV nonavalent vaccine (recombinant), HPV quadrivalent vaccine (recombinant), Hycamtin (topotecan hydrochloride), Hyper-CVAD, Ibrance (palbociclib), ibritumomab tiuxetan, ibrutinib, ICE, Iclusig (ponatinib hydrochloride), Idamycin (idarubicin hydrochloride), idarubicin hydrochloride, idelalisib, Ifex (ifosfamide), ifosfamide, Ifosfamidam (ifosfamide), imatinib mesylate, Ibruvica (ibrutinib) , imiquimod, Inlyta (axitinib), interferon alfa-2b, recombinant, Intron A (recombinant interferon alfa-2b), iodine I131 tositumomab and tositumomab, ipilimumab, Iressa (gefitinib), irinotecan hydrochloride, Istodax (romidepsin), ixabepilone, Ixempra (ixabepilone), Jakafi (ruxolitinib), Jevtana (cabazitaxel), Kadcyla (adotrastuzumab emtansine), Keoxifen (raloxifene hydrochloride), Kepivance (parif Hermin), Keytruda (pembrolizumab), Kyprolis (carfilzomib), lanreotide acetate, lapatinib ditosylate, lenalidomide, lenvatinib mesylate, Lenvima (lenvatinib mesylate), letrozole, leucovorin calcium, Leukelan (chlorambucil), leuprorelin acetate, Levran (aminolevulinic acid), Linfolidine (chlorambucil), Lipodox (liposomal doxorubicin hydrochloride), liposomal cytarabine, lomustine, Lupron (leuprorelin acetate), Lupron Depot (leuprorelin acetate),Lupron Depot-Ped (leuprorelin acetate), Lupron Depot-3 Month (leuprorelin acetate), Lupron Depot-4 Month (leuprorelin acetate), Lynparza (olaparib), Marchibo (vincristine sulfate liposome), Matulane (procarbazine hydrochloride), mechlorethamine hydrochloride, Megas (megestrol acetate), megestrol acetate, Mekinist (trametinib), mercaptopurine, mesna, Mesnex (mesna), metazolastone (temozolomide), methotrexate, methotrexate LPF (methotrexate), Mexate (methotrexate), Mexate AQ (methotrexate), mitomycin C, mitoxantrone hydrochloride, Mitozitrex (mitomycin C), MOPP, Mozobil (plelixafor), Mustagen (mechlorethamine hydrochloride), Mutamycin (mitomycin C), Myleran (busulfan), Mirosal (azacitidine), Milotarg (gemtuzumab ozogamicin), nanoparticle paclitaxel (paclitaxel albumin-stabilized nanoparticle formulation), Navelbine (vinorelbine tartrate), nelarabine, Neosal (cyclophosphamide), netupitant, and palonosetron hydrochloride , Neupogen (filgrastim), Nexavar (sorafenib tosylate), nilotinib, nivolumab, Nolvadex (tamoxifen citrate), N-plate (romiplostim), obinutuzumab, Odomzo (sonidegib), OEPA, ofatumumab, OFF, olaparib, omacetaxine mepescoxin, Oncaspar (pegaspargase), ondansetron hydrochloride, Ontac (denileukin diftitox), Opdivo (nivolumab), OPPA, oxaliplatin, paclitaxel, Paclitaxel albumin-stabilized nanoparticle formulation, PAD, palbociclib, palifermin, palonosetron hydrochloride, palonosetron hydrochloride and netupitant, pamidronate disodium, panitumumab, panobinostat, Paraplat (carboplatin), Paraplatin (carboplatin), pazopanib hydrochloride, pegaspargase, peginterferon alfa-2b, pegIntron (peginterferon alfa-2b), pembrolizumab, pemetrexed disodium, Perjeta (pertuzumab),Pertuzumab, Platinol (cisplatin), Platinol AQ (cisplatin), plerixafor, pomalidomide, Pomalyst (pomalidomide), ponatinib hydrochloride, pralatrexate, prednisone, procarbazine hydrochloride, Proleukin (aldesleukin), Prolia (denosumab), Promacta (eltrombopag olamine), Provenzi (sipulecol-T), Purinetol (mercaptopurine), Prixan (mercaptopurine), radium-223 dichloride, raloxifene hydrochloride, ramucirumab, rasburicase, R-CHO P, R-CVP, recombinant human papillomavirus (HPV) bivalent vaccine, recombinant human papillomavirus (HPV) nonavalent vaccine, recombinant human papillomavirus (HPV) quadrivalent vaccine, recombinant interferon alpha-2b, regorafenib, R-EPOCH, Revlimid (lenalidomide), Rheumatrex (methotrexate), Rituxan (rituximab), rituximab, romidepsin, romiplostim, rubidomycin (daunorubicin hydrochloride), ruxolitinib phosphate, Sclerosol intrapleural aerosol (talc), siltuximab Mab, Sipulecel-T, Somatuline Depot (lanreotide acetate), sonidegib, sorafenib tosylate, Sprycel (dasatinib), Stanford V, sterile talc powder (talc), Steritalc (talc), Stivarga (regorafenib), sunitinib malate, Sutent (sunitinib malate), Silatron (peginterferon alfa-2b), Silvant (siltuximab), Sinovir (thalidomide), Synribo (omacetaxine mepescoxate), TAC, Tafinlar (dabrafenib), talc, tamoxifen Enoate, Tarabin PFS (cytarabine), Tarceva (erlotinib hydrochloride), Targretin (bexarotene), Tasigna (nilotinib), Taxol (paclitaxel), Taxotere (docetaxel), Temodar (temozolomide), temozolomide, temsirolimus, thalidomide, Talomide (thalidomide), thiotepa, Toposar (etoposide), topotecan hydrochloride, toremifene, Torisel (temsirolimus), tositumomab and iodine I131 tositumomab, Totect (dexrazoxane hydrochloride), TPF, trametinib, trastuzumab,Treanda (bendamustine hydrochloride), Trisenox (arsenic trioxide), Tykerb (lapatinib ditosylate), Unituxin (dinutuximab), vandetanib, VAMP, Vectibix (panitumumab), VeIP, Velban (vinblastine sulfate), Velcade (bortezomib), Belsar (, Vinblastine sulfate), vemurafenib, Bepesid (etoposide), Viadur (leuprolide acetate), Vidaza (azacitidine), vinblastine sulfate, Vincasar PFS (vincristine sulfate), vincristine sulfate, vincristine sulfate liposomal, vinorelbine tartrate, VIP, vismodegib, voraxazepam (glucarpidase), vorinostat, Votrient (pazopanib hydrochloride), Wellcovorin (leucovorin calcium), Xalkori (crizotinib), Xeloda (capecitabine), Zeliri, Xerox, Zeba (denosumab), Zofigo (radium-223 dichloride), XTANDI (enzalutamide), Yervoy (ipilimumab), Zaltrap (dibuaflibercept), Zelboraf (vemurafenib), Zevalin (ibritumomab tiuxetan), Zincard (dexrazoxane hydrochloride), dibuaflibercept, Zofran (ondansetron hydrochloride), Zoladex (goserelin acetate), zoledronic acid, Zolinza (vorinostat), Zometa (zoledronic acid), Zydelig (idelalisib), Zykadia (ceritinib), and Zytiga (abiraterone acetate).

[0413] Growth factors useful as therapeutic agents include transforming growth factor alpha ("TGF-α"), transforming growth factor ("TGF-β"), platelet-derived growth factor ("PDGF"), fibroblast growth factors ("FGF") including FGF acidic isoforms 1 and 2, FGF basic form 2, and FGF4, 8, 9, and 10, nerve growth factors (NGFs) including NGF2.5, NGF7.0, βNGF, and neurotrophic factors, brain-derived neurotrophic factor, cartilage-derived factor, bone growth factor (BGF), basic fibroblast growth factor (BGF), and fibroblast growth factor (FGF). Factors include, but are not limited to, insulin-like growth factors (IGFs), vascular endothelial growth factor (VEGF), granulocyte colony-stimulating factor (G-CSF), insulin-like growth factors (IGFs) I and II, hepatocyte growth factor, glial neurotrophic growth factor (GDNF), stem cell factor (SCF), keratinocyte growth factor (KGF), transforming growth factors (TGFs) including TGFα, β, β1, β2, β3, skeletal growth factor, bone matrix-derived growth factor, and bone-derived growth factor, and mixtures thereof.

[0414] Immunoglobulins useful in the present disclosure include, but are not limited to, IgG, IgA, IgM, IgD, IgE, and mixtures thereof. Some preferred growth factors include VEGF (vascular endothelial growth factor), NGF (nerve growth factor), PDGF-AA, PDGF-BB, PDGF-AB, FGFb, FGFa, and BGF.

[0415] Other molecules useful as anti-cancer agents include, but are not limited to, growth hormone, leptin, leukemia inhibitory factor (LIF), tumor necrosis factors alpha and beta, endostatin, thrombospondin, bone morphogenetic protein 1, bone morphogenetic proteins 2 and 7, osteonectin, somatomedin-like peptide, and osteocalcin.

[0416] Tumor antigens can be based on specific mutations (neoepitopes) or expressed by cancer germline genes (antigens common to tumors found in multiple patients, referred to herein as "traditional cancer antigens" or "shared cancer antigens"). In some embodiments, traditional antigens are antigens known to be present in cancers or tumors in general, or in specific types of cancers or tumors. In some embodiments, traditional cancer antigens are non-mutated tumor antigens. In some embodiments, traditional cancer antigens are mutated tumor antigens.

[0417] Diagnostic agents include gases, metals, commercially available contrast agents and contrast media used in positron emission tomography (PET), computer-assisted tomography (CAT), single-photon emission computed tomography, x-ray, fluoroscopy, and magnetic resonance imaging (MRI). Examples of suitable substances that can be used as MRI contrast agents include gadolinium chelates, as well as iron, magnesium, manganese, copper, and chromium. Examples of materials useful for CAT and x-ray imaging include iodine-based materials.

[0418] Vaccines can include isolated proteins or peptides, inactivated organisms and viruses, killed organisms and viruses, genetically modified organisms or viruses, cell extracts, and RNA encoding at least one antigenic polypeptide or an immunogenic fragment thereof (e.g., an immunogenic fragment capable of inducing an immune response against the antigenic polypeptide). The active agent can be combined with an adjuvant, such as an interleukin, interferon, cytokine, cholera toxin, alum, Freund's adjuvant, etc.

[0419] Prophylactic agents include, for example, antigens of infectious agents, bacterial organisms, such as Streptococcus pneumoniae, Haemophilus influenzae, Staphylococcus aureus, Streptococcus pyrogenes, Corynebacterium diphtheriae, Listeria monocytogenes, Bacillus anthrax, Clostridium tetani, Clostridium botulinum, Clostridium perfringens, Neisseria meningitidis, Neisseria gonorrhoeae, Clostridium mutans, Pseudomonas aeruginosa, Salmonella typhi, Haemophilus influenzae, Bordetella pertussis, Francisella tularensis, Yersinia pestis, Vibrio cholerae, Legionella pneumophila, Mycobacterium tuberculosis, Mycobacterium leprae, Treponema pallidum, L. interrogans, Borrelia burgdorferi, Campylobacter jejuni, Antigens of viruses, such as human metapneumovirus (hMPV), human parainfluenza virus (hPIV) types 1, 2, and 3 (hPIV1, hPIV2, and hPIV3, respectively), respiratory syncytial virus (RSV), measles virus (MeV), coronaviruses (e.g., MERS-CoV, SARS-CoV, SARS-CoV2, HCoV-OC43, HCoV-229E, HCoV-NL63, HCoV-NL, HCoV-NH, and HCoV-HKU1), and poxviruses Viruses (e.g., smallpox, monkeypox), African swine virus, influenza A and B, HIV, varicella-zoster, herpes simplex types 1 and 2, cytomegalovirus, Epstein-Barr virus, rotavirus, rhinovirus, adenovirus, papillomavirus, poliovirus, mumps, rabies, rubella, coxsackievirus, equine encephalitis, Japanese encephalitis, yellow fever, Rift Valley fever, hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis D virus, hepatitis E virus Antigens may include fungal, protozoan, and parasitic antigens, such as Cryptococcus neoformans, Histoplasma capsulatum, Candida albicans, Candida tropicalis, Nocardia asteroides, Rickettsia rickettsii, Salmonella typhi, Mycoplasma pneumoniae, Chlamydia psittaci, Chlamydia trachomatis, Plasmodium falciparum, Trypanosoma brucei, Entamoeba histolytica, Toxoplasma gondii, Trichomonas protozoa, Schistosoma mansoni, etc. These antigens can be in the form of killed whole organisms, peptides, proteins, glycoproteins, carbohydrates, or combinations thereof.

[0420] In certain embodiments, the active agent is a polynucleotide. Polynucleotides or oligonucleotides that can be introduced according to the methods of the present invention include all types of DNA, cDNA, and RNA sequences. For example, the polynucleotide can be double-stranded DNA, single-stranded DNA, complex DNA, encapsulated DNA, naked RNA, encapsulated RNA, messenger RNA (mRNA), tRNA, short interfering RNA (siRNA), double-stranded RNA (dsRNA), microRNA (miRNA), antisense RNA (asRNA), self-amplifying mRNA (saRNA), guide RNA (gRNA), cRNA, and combinations thereof. The polynucleotide can be a DNA construct, such as an expression vector, encoding a desired gene product (e.g., a gene product homologous or heterologous to the target gene product).

[0421] In some embodiments, RNA (e.g., mRNA) can be used to induce a balanced immune response to infectious agents. In some embodiments, RNA (e.g., mRNA) can be used to induce a balanced immune response to infectious agents, such as metapneumoviruses, such as human metapneumovirus (hMPV), parainfluenza viruses, such as human parainfluenza viruses (hPIV) types 1, 2, and 3 (hPIV1, hPIV2, and hPIV3, respectively), respiratory syncytial virus (RSV), measles virus (MeV), coronaviruses (e.g., MERS-CoV, SARS-CoV, SARS-CoV2, HCoV-OC43, HCoV-229E, HCoV-NL63, HCoV-NL, HCoV-NH, HCoV-H), or other viruses. In some embodiments, RNA (e.g., mRNA) may be used to induce a balanced immune response against viruses such as HIV, poxviruses (e.g., smallpox, monkeypox), African swine virus, influenza A and B, HIV, varicella-zoster, herpes simplex types 1 and 2, cytomegalovirus, Epstein-Barr virus, rotavirus, rhinovirus, adenovirus, papillomavirus, poliovirus, mumps, rabies, rubella, coxsackievirus, equine encephalitis, Japanese encephalitis, yellow fever, Rift Valley fever, and hepatitis A, B, C, D, and E viruses. In some embodiments, RNA (e.g., mRNA) may be used to induce a balanced immune response against respiratory viruses. As used herein, the term "respiratory virus" refers to viruses that cause respiratory disease. They are negative-sense, single-stranded RNA viruses of the Paramyxoviridae family, such as human metapneumovirus (hMPV), human parainfluenza virus (hPIV) types 1, 2, and 3 (hPIV1, hPIV2, and hPIV3, respectively), respiratory syncytial virus (RSV), and measles virus (MeV). Another example of a respiratory virus is coronavirus. Coronaviruses are enveloped viruses with a positive-sense, single-stranded RNA genome and a nucleocapsid with helical symmetry. Coronaviruses are a type of virus that belong to the Coronaviridae family, Coronaviridae subfamily, and Coronavirinae subfamily of the Nidovirales order.

[0422] Representative examples of beta coronaviruses include, but are not limited to, envecoviruses (e.g., beta coronavirus 1, human coronavirus OC43, Chinese murine coronavirus HKU24, human coronavirus HKU1, and mouse coronavirus), hybecoviruses (e.g., Bat Hp-betacoronavirus Zhejiang2013), merbecoviruses (e.g., hedgehog coronavirus 1, Middle East respiratory syndrome-associated coronavirus (MERS-CoV), pipistrelle bat coronavirus HKU5, and Cryonycteris bat coronavirus HKU4), nobecoviruses (e.g., leucette bat coronavirus GCCDC1 and leucette bat coronavirus HKU9), and sarbecoviruses (e.g., severe acute respiratory syndrome coronavirus (SARS-CoV), severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)).

[0423] Representative examples of gamma coronaviruses include, but are not limited to, segakoviruses (eg, beluga coronavirus SQ1) and igakoviruses (eg, avian coronavirus (IBV)).

[0424] Representative examples of delta coronaviruses include, but are not limited to, anddecoviruses (e.g., Wigeon coronavirus HKU20), burdecoviruses (e.g., brown-eared brown coronovirus HKU11, porcine coronavirus HKU15 (PorCoV HKU15)), munia coronavirus HKU13, white-eye coronavirus HKU16), herdecoviruses (e.g., black-crowned night heron coronavirus HKU19), and murdecoviruses (e.g., stag beetle coronavirus HKU21).

[0425] In some embodiments, the coronavirus is a human coronavirus. Representative examples of human coronaviruses include, but are not limited to, human coronavirus 229E (HCoV-229E), human coronavirus OC43 (HCoV-OC43), human coronavirus HKU1 (HCoV-HKU1), human coronavirus NL63 (HCoV-NL63), severe acute respiratory syndrome coronavirus (SARS-CoV), severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and Middle East respiratory syndrome-related coronavirus (MERS-CoV).

[0426] In some embodiments, the RNA (e.g., mRNA) polynucleotide having an open reading frame is

[0427] At least one (e.g., at least two, three, four, or five) human metapneumovirus (hMPV) antigen polypeptide, human parainfluenza virus (hPIV) types 1, 2, and 3 (hPIV1, hPIV2, and hPIV3, respectively) antigen polypeptide, respiratory syncytial virus (RSV) antigen polypeptide, measles virus (MeV) antigen polypeptide, varicella-zoster antigen polypeptide, influenza virus antigen polypeptide, herpes simplex virus 1 (HSV1) antigen polypeptide, herpes simplex virus 2 (HSV2) antigen polypeptide, poxvirus (e.g., smallpox, monkeypox) antigen polypeptide, African swine virus antigen polypeptide, cytomegalovirus antigen polypeptide, Epstein-Barr virus antigen polypeptide The polypeptides encode antigens of rabies, flu, flu-like viruses, rabies virus, flu-like viruses ...As used herein, the use of the term "antigenic polypeptide" refers to an immunogenic fragment of an antigenic polypeptide (an immunogenic fragment that induces (or is capable of inducing) an immune response to human metapneumovirus (hMPV), human parainfluenza virus (hPIV) types 1, 2, and 3 (hPIV1, hPIV2, and hPIV3, respectively), respiratory syncytial virus (RSV), measles virus (MeV), varicella-zoster virus, influenza virus, herpes simplex virus type 1 (HSV1), herpes simplex virus 2 (HSV2), poxvirus (such as smallpox and monkeypox), and avian influenza virus. The virus may be an African swine virus, cytomegalovirus, Epstein-Barr virus, rotavirus, rhinovirus, adenovirus, papillomavirus, poliovirus, mumps, rabies, rubella, coxsackievirus, equine encephalitis, Japanese encephalitis, yellow fever, Rift Valley fever, hepatitis A, B, C, D, E virus, or coronavirus (e.g., MERS-CoV, SARS-CoV, SARS-CoV2, HCoV-OC43, HCoV-229E, HCoV-NL63, HCoV-NL, HCoV-NH, HCoV-HKU1), or a combination thereof.

[0428] In some embodiments, the agent is an RNA (e.g., mRNA) capable of inducing a balanced immune response, including both cellular and humoral immunity, against hMPV, PIV, RSV, MeV, and / or a coronavirus (e.g., MERS-CoV, SARS-CoV, SARS-CoV2, HCoV-OC43, HCoV-229E, HCoV-NL63, HCoV-NL, HCoV-NH, and / or HCoV-HKU1), or any combination of two or more of the foregoing viruses, without risking the possibility of insertional mutagenesis.

[0429] In some embodiments, the nucleic acids disclosed herein may contain at least one chemically modified nucleotide. In some embodiments, the at least one chemically modified nucleotide comprises a chemically modified nucleic acid base, a chemically modified ribose, a chemically modified phosphodiester linkage, or a combination thereof.

[0430] In one embodiment, at least one chemically modified nucleotide is a chemically modified nucleobase.

[0431] In one embodiment, the chemically modified nucleobase is 5-formylcytidine (5fC), 5-methylcytidine (5meC), 5-methoxycytidine (5moC), 5-hydroxycytidine (5hoC), 5-hydroxymethylcytidine (5hmC), 5-formyluridine (5fU), 5-methyluridine (5-meU), 5-methoxyuridine (5moU), 5-carboxymethylesteruridine (5camU), pseudouridine (Ψ), N1-methylpseudouridine (me 1 Ψ), N6-methyladenosine (me 6 A), or thienoguanosine (thG).

[0432] In some embodiments, the chemically modified nucleobase is 5-methoxyuridine (5moU). In some embodiments, the chemically modified nucleobase is pseudouridine (Ψ). In some embodiments, the chemically modified nucleobase is N1-methylpseudouridine (me 1 Ψ).

[0433] The structures of these modified nucleobases are shown below: [ka]

[0434] In one embodiment, at least one chemically modified nucleotide is a chemically modified ribose.

[0435] In one embodiment, the chemically modified ribose is selected from 2'-O-methyl (2'-O-Me), 2'-fluoro (2'-F), 2'-deoxy-2'-fluoro-beta-D-arabinonucleic acid (2'F-ANA), 4'-S, 4'-SFANA, 2'-azido, UNA, 2'-O-methoxyethyl (2'-O-ME), 2'-O-allyl, 2'-O-ethylamine, 2'-O-cyanoethyl, locked nucleic acid (LAN), methylene-cLAN, N-MeO-amino BNA, or N-MeO-aminooxy BNA. In one embodiment, the chemically modified ribose is 2'-O-methyl (2'-O-Me). In one embodiment, the chemically modified ribose is 2'-fluoro (2'-F).

[0436] The structures of these modified riboses are shown below: [ka]

[0437] In one embodiment, at least one chemically modified nucleotide is a chemically modified phosphodiester linkage.

[0438] In one embodiment, the chemically modified phosphodiester linkage is selected from phosphorothioate (PS), boranophosphate, phosphodithioate (PS2), 3',5'-amide, N3'-phosphoramidate (NP), phosphodiester (PO), or 2',5'-phosphodiester (2',5'-PO). In one embodiment, the chemically modified phosphodiester linkage is phosphorothioate.

[0439] The structures of these modified phosphodiester linkages are shown below: [ka]

[0440] In some embodiments, the mRNA may include a heterologous 5' untranslated region (5'UTR).

[0441] In some embodiments, the mRNA may include a heterologous 3' untranslated region (3'UTR).

[0442] method Depending on the type of active agent, the composition of the present invention can be used as a composition for the diagnosis, prevention, and treatment of diseases or disorders (e.g., infection, cancer, pain, depression, inflammatory diseases, intestinal diseases, brain diseases, allergic diseases, arrhythmia, hypertension, eye diseases, endocrine diseases, and cardiovascular diseases).

[0443] Described herein are methods for delivering an active agent (e.g., a polynucleotide) into an organ, tissue, and / or cell, comprising introducing into the organ, tissue, and / or cell a composition or nanoparticle described herein comprising a compound of Formula Y, I, Ia, II, III, IV, V, or any combination thereof, and the active agent.

[0444] Described herein are methods for diagnosing, treating, or preventing disease, comprising administering to a subject in need thereof an effective amount of a composition or nanoparticles described herein comprising a compound of Formula Y, I, Ia, II, III, IV, V, or any combination thereof, and an active agent.

[0445] Described herein are methods for treating or preventing an infection, comprising administering to a subject in need thereof an effective amount of a composition or nanoparticles described herein comprising a compound of Formula Y, I, Ia, II, III, IV, V, or any combination thereof, and an active agent.

[0446] Described herein are methods for treating or preventing a respiratory infection, comprising administering to a subject in need thereof an effective amount of a composition or nanoparticles described herein comprising a compound of Formula Y, I, Ia, II, III, IV, V, or any combination thereof, and an active agent.

[0447] Also described herein are methods for inducing an immune response against a virus, comprising administering to a subject in need thereof an effective amount of a composition or nanoparticles described herein comprising a compound of Formula Y, I, Ia, II, III, IV, V, or any combination thereof, and an active agent.

[0448] Also described herein is a method of inducing an immune response against a respiratory virus, comprising administering to a subject in need thereof an effective amount of a composition or nanoparticles comprising a compound of Formula Y, I, Ia, II, III, IV, V, or any combination thereof, and an active agent.

[0449] In some embodiments, provided herein are methods for delivering a polynucleotide. In some embodiments, provided herein are methods for delivering a polynucleotide (e.g., mRNA) to provide expression of the polynucleotide (and translation to produce a protein) in a cell. In some embodiments, provided herein are methods for delivering a polynucleotide (e.g., mRNA) to induce an immune response in a subject.

[0450] In some embodiments, the method of delivery of the active agent can introduce a polynucleotide into a cell or tissue and express the protein encoded by the polynucleotide in vivo. In some embodiments, delivery of the active agent (e.g., a polynucleotide) can correct a defect caused by a deficiency of that polynucleotide in the cell or tissue.

[0451] In some embodiments, active agent delivery methods can introduce polynucleotides into cells or tissues to modulate (e.g., turn off or decrease / increase) the expression of specific genes. For example, delivery of antisense mRNA or shRNA to tumor cells can modulate the expression of the protein encoded by the mRNA or shRNA.

[0452] In some embodiments, the active agent delivery method can introduce polynucleotides into cells or tissues for cellular reprogramming, where the polynucleotides can be used to regulate cellular behavior by expressing transcription factors or growth factors.

[0453] In some embodiments, the method of delivery of the active agent can introduce a polynucleotide into cells or tissues in vivo, for example, by introducing a "suicide gene" or drug susceptibility gene into tumor cells, enabling the cells to express a prodrug-activating enzyme (e.g., herpes simplex virus thymidine kinase) that can kill the tumor cells when exposed to a prodrug (e.g., acyclovir, ganciclovir, valciclovir, or famciclovir).

[0454] In some embodiments, methods for delivering active agents can introduce polynucleotides into cells or tissues to encode polypeptides that elicit a specific immune response against target cells. In some embodiments, the target cells can be cancer cells, infected cells, or combinations thereof. In some embodiments, the polynucleotides can encode infectious antigen polypeptides, tumor antigens, cytokines (e.g., interferons, interleukins, colony-stimulating factors), monoclonal antibodies, or antibody fragments (single-chain variable fragments (scFv), fragment antigen-binding ((Fab')2), intrabodies, nanobodies), or any combination thereof.

[0455] In some aspects, disclosed herein are methods of treating cancer, comprising administering to a subject in need thereof an effective amount of a composition or nanoparticles described herein comprising a compound of Formula Y, I, Ia, II, III, IV, V, or any combination thereof, and an active agent.

[0456] In some embodiments, the method of delivery of the active agent comprises introducing a polynucleotide into specific cells or tissues ex vivo, expressing the polypeptide encoded by the polynucleotide on the specific cells or tissues to generate chimeric cells or tissues, and administering the chimeric cells or tissues to a subject for tumor treatment. In some embodiments, the specific cells comprise CAR T cells, CAR-NK cells, or other cells obtained from the subject that can be used for immunotherapy.

[0457] In some embodiments, the subject is a mammal. In some embodiments, the mammal is a human. In some embodiments, the subject is a veterinary patient.

[0458] In some embodiments, the compositions herein are used to treat both localized and metastatic tumors.

[0459] In some embodiments, the compositions and methods described herein are useful for treating or preventing metastasis or recurrence of cancer. In some embodiments, the compositions and methods described herein are useful for preventing recurrence of resected solid tumors. In some embodiments, the compositions and methods described herein are useful for preventing metastasis of resected solid tumors.

[0460] In one aspect, the methods described herein are useful in the treatment of cancer, particularly cancers such as melanoma, lung cancer (including lung adenocarcinoma, basal cell carcinoma, squamous cell carcinoma, large cell carcinoma, bronchioloalveolar carcinoma, bronchiolar carcinoma, non-small cell carcinoma, small cell carcinoma, mesothelioma), breast cancer (including ductal carcinoma, lobular carcinoma, inflammatory breast cancer, clear cell carcinoma, mucinous carcinoma, serous cavity breast carcinoma), colorectal cancer (colon cancer, rectal cancer, colorectal adenocarcinoma), anal cancer, pancreatic cancer (including pancreatic adenocarcinoma, pancreatic islet cell carcinoma, neuroendocrine tumors), prostate cancer, prostate adenocarcinoma, ovarian cancer (including serous tumors, endometrioid tumors, mucinous cystadenocarcinoma, ovarian epithelial carcinoma including sex cord-stromal tumors or surface epithelial-stromal tumors), ), liver and bile duct cancer (including hepatocellular carcinoma, cholangiocarcinoma, and hemangioma), esophageal cancer (including esophageal adenocarcinoma and squamous cell carcinoma), oral and oropharyngeal squamous cell carcinoma, salivary gland adenoid cystic carcinoma, bladder cancer, bladder carcinoma, uterine cancer (including endometrial adenocarcinoma, eye cancer, uterine papillary serous carcinoma, uterine clear cell carcinoma, uterine sarcoma, leiomyosarcoma, and mixed Müllerian tumor), glioma, glioblastoma, medulloblastoma, other brain tumors, kidney cancer (including renal cell carcinoma, clear cell carcinoma, and Wilms' tumor), head and neck cancer (including squamous cell carcinoma), stomach cancer (gastric cancer, gastric adenocarcinoma, and gastrointestinal stromal tumor), testicular cancer, germ cell tumors, and neuroendocrine tumors , cervical cancer, carcinoids of the digestive tract, breast, and other organs, signet ring cell carcinoma, mesenchymal tumors (including sarcomas), fibrosarcoma, hemangioma, hemangiomatosis, hemangiopericytoma, pseudoangiomatous stromal hyperplasia, myofibroblastoma, fibromatosis, inflammatory myofibroblastic tumor, lipoma, angiolipoma, granular cell tumor, neurofibroma, schwannoma, angiosarcoma, liposarcoma, rhabdomyosarcoma, osteosarcoma, leiomyoma, leiomyosarcoma, skin (including melanoma), cervix, retinoblastoma, head and neck cancer, pancreas, brain, thyroid, testis, kidney, bladder, soft tissue, adrenal gland, urethra, and penile cancer, myxosarcoma, chondrosarcoma, and osteosarcoma. Used to treat chordoma, malignant fibrous histiocytoma, lymphangiosarcoma, mesothelioma, squamous cell carcinoma, epidermoid carcinoma, malignant skin adnexal tumor, adenocarcinoma, hepatocellular carcinoma, hepatocellular carcinoma, renal cell carcinoma, adrenal gland tumor, cholangiocarcinoma, transitional cell carcinoma, choriocarcinoma, seminoma, embryonal cell carcinoma, anaplastic glioma, glioblastoma multiforme, neuroblastoma, medulloblastoma, malignant meningioma, malignant neurilemmoma, neurofibrosarcoma, parathyroid carcinoma, medullary thyroid carcinoma, bronchial carcinoid, pheochromocytoma, pancreatic islet cell carcinoma, malignant carcinoid, malignant paraganglioma, melanoma, Merkel cell tumor, cystosarcoma phyllodes, salivary gland carcinoma, thymic carcinoma, and vaginal carcinoma.

[0461] In some embodiments, the compositions and methods described herein are useful for treating or preventing cancer. In some cases, the cancer is a circulating cancer cell (circulating tumor cell). In some cases, the cancer is a metastatic cancer cell.

[0462] In some embodiments, the method further comprises administering an additional therapeutic agent, hi some embodiments, the additional therapeutic agent comprises an additional immunotherapeutic agent or an anti-tumor agent.

[0463] Also disclosed herein is a method of treating a disease or condition, such as an inflammatory disorder (including autoimmune diseases) or a lymphoproliferative disorder, comprising administering to a subject in need thereof an effective amount of a compound, combination of compounds, or composition provided herein, or a pharmaceutically acceptable form thereof, or a pharmaceutical composition provided herein.

[0464] Further disclosed herein is a method of treating a disease or condition, such as an inflammatory disorder (including autoimmune diseases) or a lymphoproliferative disorder, comprising administering to a subject in need thereof an effective amount of a compound, combination of compounds, or composition provided herein, or a pharmaceutically acceptable form thereof, or a pharmaceutical composition provided herein.

[0465] In one embodiment, provided herein is a method for treating inflammatory disorders, including autoimmune diseases, in a subject. The method comprises administering to the subject a therapeutically effective amount of a compound, combination of compounds, or composition provided herein, or a pharmaceutically acceptable form thereof, or a pharmaceutical composition provided herein. Examples of autoimmune diseases include acute disseminated encephalomyelitis (ADEM), Addison's disease, antiphospholipid syndrome (APS), aplastic anemia, autoimmune hepatitis, autoimmune skin diseases, celiac disease, Crohn's disease, diabetes mellitus (type 1), Goodpasture's syndrome, Graves' disease, Guillain-Barré syndrome (GBS), Hashimoto's disease, systemic lupus erythematosus, multiple sclerosis, myasthenia gravis, eyelid myoclonus syndrome (OMS), optic neuritis, ordo's thyroiditis, urticaria major, and multiple sclerosis. Diseases include, but are not limited to, arthritis, primary biliary cirrhosis, psoriasis, rheumatoid arthritis, Reiter's syndrome, Takayasu's arteritis, temporal arteritis (also called "giant cell arteritis"), warm autoimmune hemolytic anemia, Wegener's granulomatosis, alopecia universalis (e.g., inflammatory alopecia), Chagas' disease, chronic fatigue syndrome, dysautonomia, endometriosis, hidradenitis suppurativa, interstitial cystitis, myotonia neuropathica, sarcoidosis, scleroderma, ulcerative colitis, vitiligo, and vulvodynia. Other diseases include bone resorption disorders and thrombosis.

[0466] Inflammation has many forms, including, but not limited to, acute, adhesive, atrophic, catarrhal, chronic, cirrhotic, diffuse, disseminated, exudative, fibrotic, fibrosing, focal, granulomatous, hyperplastic, hypertrophic, interstitial, metastatic, necrotic, obstructive, parenchymal, plastic, proliferative, pseudomembranous, suppurative, sclerosing, serous, serous, simple, specific, subacute, suppurative, toxic, traumatic, and / or ulcerative inflammation.

[0467] Exemplary inflammatory conditions include inflammation associated with acne, anemia (e.g., aplastic anemia, hemolytic autoimmune anemia), asthma, arteritis (e.g., polyarteritis, temporal arteritis, periarteritis nodosa, Takayasu's arteritis), arthritis (e.g., crystalline arthritis, osteoarthritis, psoriatic arthritis, gout attacks, gouty arthritis, reactive arthritis, rheumatoid arthritis, Reiter's arthritis), ankylosing spondylitis, amylose, amyotrophic lateral sclerosis, autoimmune diseases, allergies or allergic reactions, arteriosclerosis, bronchitis, bursitis, chronic prostatitis, conjunctivitis, Chagas' disease, chronic obstructive pulmonary disease, dermatomyositis, diverticulitis, and the like. inflammation, diabetes (e.g., type 1 diabetes, type 2 diabetes), skin diseases (e.g., psoriasis, eczema, burns, dermatitis, pruritus (itching)), endometriosis, Guillain-Barré syndrome, infection, ischemic heart disease, Kawasaki disease, glomerulonephritis, gingivitis, hypersensitivity, headache (e.g., migraine, tension headache), intestinal obstruction (e.g., postoperative ileus, ileus during sepsis), idiopathic thrombocytopenic purpura, interstitial cystitis (bladder pain syndrome), gastrointestinal disorders (e.g., peptic ulcer, regional enteritis, diverticulitis, gastrointestinal bleeding, eosinophilic gastrointestinal disorders (e.g., eosinophilic esophagitis, eosinophilic gastritis, eosinophilic gastroenteritis, eosinophilic colitis), gastritis, Diarrhea, gastroesophageal reflux disease (GORD, or its synonym GERD), inflammatory bowel disease (IBD) (e.g., Crohn's disease, ulcerative colitis, collagenous colitis, lymphocytic colitis, ischemic colitis, transformation colitis, Behçet's syndrome, indeterminate bowel disease (IBS)), lupus, multiple sclerosis, myasthenia gravis, myocardial ischemia, nephrotic syndrome, pemphigus vulgaris, pernicious anemia, peptic ulcer, polymyositis, primary biliary cirrhosis, neuroinflammation associated with brain diseases (e.g., Parkinson's disease, Huntington's disease, Alzheimer's disease), prostatitis, chronic inflammation associated with cranial radiation injury, In certain embodiments, the inflammatory disorder is selected from arthritis (e.g., rheumatoid arthritis), inflammatory bowel disease, inflammatory bowel syndrome, asthma, psoriasis, endometriosis, interstitial cystitis, and prostatitis.In certain embodiments, the inflammatory condition is an acute inflammatory condition (e.g., inflammation resulting from infection). In certain embodiments, the inflammatory condition is a chronic inflammatory condition (e.g., conditions resulting from asthma, arthritis, inflammatory bowel disease). These compounds are also useful for treating inflammation associated with trauma and non-inflammatory muscle pain.

[0468] Immune disorders, such as autoimmune disorders, include arthritis (including rheumatoid arthritis, spondyloarthropathy, gouty arthritis, osteoarthritis, systemic lupus erythematosus, Sjögren's syndrome, ankylosing spondylitis, undifferentiated spondylitis, Behçet's disease, hemolytic autoimmune anemia, multiple sclerosis, amyotrophic lateral sclerosis, amylosis, acute shoulder pain, psoriasis, and juvenile arthritis), asthma, and arteriosclerosis. osteoporosis, bronchitis, tendonitis, bursitis, skin diseases (psoriasis, eczema, burns, dermatitis, pruritus (itching), etc.), nocturnal enuresis, eosinophilic disease, gastrointestinal disorders (e.g., selected from peptic ulcer, regional enteritis, diverticulitis, gastrointestinal bleeding, eosinophilic gastrointestinal disorders (e.g., eosinophilic esophagitis, eosinophilic gastritis, eosinophilic gastroenteritis, eosinophilic colitis)), gastritis, diarrhea, gastroesophageal reflux disease (GORD) or its synonym GERD), inflammatory bowel disease (IBD) (e.g., Crohn's disease, ulcerative colitis, collagenous colitis, lymphocytic colitis, ischemic colitis, transformation colitis, Behcet's syndrome, indeterminate colitis), and inflammatory bowel syndrome (IBS), relapsing polychondritis (e.g., atrophic polychondritis, generalized polychondromalacia), and diseases that improve with gastrokinetic agents (intestinal obstruction, postoperative ileus, ileus in sepsis, gastroesophageal reflux disease (GORD, or its synonym GERD)), eosinophilic esophagitis, gastroparesis such as diabetic gastroparesis, food intolerances and food allergies, and other functional bowel disorders such as non-ulcer dyspepsia (NUD) and non-cardiac chest pain (NCCP, including costochondritis).

[0469] In one embodiment, provided herein is a method of treating an infection in a subject caused by an infectious agent, the method comprising administering to the subject a therapeutically effective amount of a compound, combination of compounds, or composition provided herein, or a pharmaceutically acceptable form thereof, or a pharmaceutical composition provided herein.Infectious agents include bacterial organisms such as Streptococcus pneumoniae, Haemophilus influenzae, Staphylococcus aureus, Streptococcus pyrogenes, Corynebacterium diphtheriae, Listeria monocytogenes, Bacillus anthrax, Clostridium tetani, Clostridium botulinum, Clostridium perfringens, Neisseria meningitidis, Neisseria gonorrhoeae, Clostridium mutans, Pseudomonas aeruginosa, Salmonella typhi, Haemophilus influenzae, Bordetella pertussis, Francisella tularensis, Yersinia pestis, Vibrio cholerae, Legionella pneumophila, Mycobacterium tuberculosis, Mycobacterium leprae, Treponema pallidum, Rhodeus interrogans, Borrelia burgdorferi, Campylobacter jejuni, and viruses such as Metapneumococcal Viruses, such as human metapneumovirus (hMPV), parainfluenza viruses, such as human parainfluenza virus (hPIV) types 1, 2, and 3 (hPIV1, hPIV2, and hPIV3, respectively), respiratory syncytial virus (RSV), measles virus (MeV), coronaviruses (e.g., MERS-CoV, SARS-CoV, SARS-CoV2, HCoV-OC43, HCoV-229E, HCoV-NL63, HCoV-NL, HCoV-NH, and HCoV-HKU1), poxviruses, and Viruses (e.g., smallpox, monkeypox), African swine virus, influenza A and B, human immunodeficiency virus (HIV), varicella-zoster, herpes simplex types 1 and 2, cytomegalovirus, Epstein-Barr virus, rotavirus, rhinovirus, adenovirus, papillomavirus, poliovirus, mumps, rabies, rubella, coxsackievirus, equine encephalitis, Japanese encephalitis, yellow fever, Rift Valley fever, hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis D virus, hepatitis E virus Viruses, fungi, protozoa, and parasites, including, but not limited to, Cryptococcus neoformans, Histoplasma capsulatum, Candida albicans, Candida tropicalis, Nocardia asteroides, Rickettsia rickettsii, Salmonella typhi, Mycoplasma pneumoniae, Chlamydia psittaci, Chlamydia trachomatis, Plasmodium falciparum, Trypanosoma brucei, Entamoeba histolytica, Toxoplasma gondii, Trichomonas protozoa, and Schistosoma mansoni.

[0470] In some aspects, the infection can be a coronavirus infection. In one embodiment, the coronavirus infection is an infection of the upper and / or lower respiratory tract. The "upper respiratory tract" includes the mouth, nose, sinuses, middle ear, throat, larynx, and trachea. The "lower respiratory tract" includes the bronchial tubes (bronchi) and lungs (bronchi, bronchioles, alveoli), and interstitial tissue of the lungs. In another embodiment, the coronavirus infection is an infection of the gastrointestinal tract, where "gastrointestinal tract" includes any region of the tract from the mouth to the anus, including the mouth, esophagus, stomach, and intestines.

[0471] In yet another embodiment, the coronavirus infection is a kidney infection.

[0472] It is understood and contemplated herein that the coronavirus infections disclosed herein can cause pathological conditions associated with coronavirus infection, referred to herein as "coronavirus diseases." In some embodiments, the coronavirus infection is selected from the common cold, pneumonia, pneumonitis, bronchitis, severe acute respiratory syndrome (SARS), coronavirus infection 2019 (COVID-2019), Middle East respiratory syndrome (MERS), sinusitis, porcine diarrhea, porcine epidemic diarrhea, avian infectious bronchitis, otitis media, and pharyngitis. In some embodiments, the coronavirus infection is the common cold. In some embodiments, the coronavirus infection is selected from SARS, COVID-19, and MERS. In certain embodiments, the coronavirus infection is COVID-19. In other specific embodiments, the coronavirus infection is IBV, PorCoV HKU15, or PEDV.

[0473] Most patients diagnosed with SARS have been previously healthy adults aged 25 to 70 years. A few suspected cases of SARS have been reported among children under the age of 15. The case fatality rate for sick people who meet the current World Health Organization definition of a suspected SARS case is approximately 3%.

[0474] Other symptoms associated with coronavirus infection are described in Graninski & Baric, 2015, J. Pathol. 235:185-195 and Cavanagh, 2005, "Coronaviridae: a review of coronaviruses and toroviruses," Coronaviruses with Special Emphasis on First Insights Concerning SARS 1, ed. By A. Schmidt, M. H. Wolff and O. Weber, Birkhauser Verlag Baser, Switzerland, each of which is incorporated herein by reference in its entirety.

[0475] The coronavirus causing the infection may be selected from an alpha coronavirus, a beta coronavirus, a gamma coronavirus, or a delta coronavirus.

[0476] Ocular diseases that may be treated in accordance with the compositions and methods disclosed herein include amoebic keratitis, fungal keratitis, bacterial keratitis, viral keratitis, blepharokeratitis, bacterial keratoconjunctivitis, viral keratoconjunctivitis, corneal dystrophic diseases, Fuchs' endothelial dystrophy, Sjogren's syndrome, Stevens-Johnson syndrome, autoimmune dry eye disease, environmental dry eye disease, corneal neovascularization diseases, prevention and treatment of corneal transplant rejection, autoimmune Includes uveitis, infectious uveitis, anterior uveitis, posterior uveitis (including toxoplasmosis), panuveitis, inflammatory diseases of the vitreous or retina, prevention and treatment of endophthalmitis, macular edema, macular degeneration, age-related macular degeneration, proliferative and non-proliferative diabetic retinopathy, hypertensive retinopathy, autoimmune diseases of the retina, primary and metastatic intraocular melanoma, other intraocular metastatic tumors, open-angle glaucoma, angle-closure glaucoma, pigmentary glaucoma, and combinations thereof.

[0477] Endocrine disorders that may be treated in accordance with the compositions and methods disclosed herein include thyroid disorders (e.g., hyperthyroidism, hypothyroidism, thyroiditis, goiter, diabetes, hypoglycemia, glucagonoma, calcium homeostasis disorders (e.g., parathyroid gland, osteoporosis, osteomalacia, rickets), sex hormone disorders (e.g., disorders of sex development, hypogonadism, pubertal disorders, menstrual function, or infertility). [Example]

[0478] The following examples are presented below to illustrate compounds, compositions, methods, and results according to the disclosed subject matter. These examples are not intended to be exhaustive of all aspects of the subject matter disclosed herein, but are intended to illustrate representative methods and results. These examples are not intended to exclude equivalents and variations of the present invention that would be apparent to one skilled in the art.

[0479] Example 1: Synthesis scheme of ionizable lipid (compound target 1) We now describe the experimental procedure for synthesizing Compound Target 1. The synthetic route for Compound 1 is shown in Figure 1. [ka]

[0480] Preparation of Compound 2: Mg (48.5 g, 1.99 mol, 3.08 equiv) and I2 (20.0 mg, 78.8 μmol) were placed in R1 (a 3.00 L three-neck round-bottom flask) under N2 at 30-40 °C. Compound 1 (125 g, 647 mmol, 113 mL, 1.00 equiv) was placed in THF (1.20 L) and placed in R2 (a 2.00 L flask) at 30-40 °C. 5.00 mL of R2 was placed in R1 at 30-40 °C. An exothermic reaction was observed, and the solution quickly lost color and began to reflux. The remaining R2 was placed in 3R1 at 0-40 °C. Stir R1 under N2 at 35 °C for 1 h. An exothermic reaction was observed, and the color changed to dark brown. The mixture was used in the following step without further purification. Compound 2 (140 g, 644 mmol, 99.5% yield) was dissolved in THF (1.20 L) and used as a dark brown liquid in the following step. [ka]

[0481] Preparation procedure for compound 3: Compound 2 (141 g, 647 mmol, 146 mL, 1.40 equiv.) in THF (500 mL) is placed in R1 (2.00 L three-neck flask). R1 is cooled to 0 °C. THF (400 mL) is placed in R2 (1.00 L barrel). Compound 2a (52.8 g, 462 mmol, 64.5 mL, 1.00 equiv.) is placed in R2. The solution in R2 is added dropwise to R1 at 0 °C. R2 is stirred at 25 °C for 16 hours. H2O (200 mL) is added to R2 (2.00 L barrel) at 25 °C. The reaction mixture is extracted with ethyl acetate (800 mL*3) at 25 °C. The reaction mixture is dried over anhydrous Na2SO4 at 25 °C. The organic phase is concentrated below 40-50 °C to obtain a residue. The residue was purified by column chromatography to give compound 3 (129 g, 565 mmol, crude) as a white solid. 1 H NMR:400MHz, CDCl3δ3.61-3.57(m,1H),1.47-1.43(m,6H),1.29-1.25(m,18H),0.91-0.87(m,6H)). [ka]

[0482] Preparation procedure for compound 4: To a solution of compound 3 (50.0 g, 219 mmol, 1.10 equiv.) and compound 3a (41.6 g, 199 mmol, 1.00 equiv.) in 500 mL of dichloromethane at 25 °C, add EDCI (49.6 g, 259 mmol, 1.30 equiv.), DIEA (103 g, 796 mmol, 139 mL, 4.00 equiv.), and DMAP (4.86 g, 39.8 mmol, 0.20 equiv.). Stir the mixture at 25 °C for 16 hours. Add 500 mL of H2O to the mixture and extract the reaction with dichloromethane (400 mL * 3) at 25 °C. Wash the reaction with saturated brine (300 mL * 3) at 25 °C. Dry the reaction with anhydrous Na2SO4 at 25 °C. The organic phase was concentrated at 40-50°C or below to obtain a residue. The residue was purified by column chromatography to obtain compound 4 (32.0 g, 78.7 mmol, yield 39.5%) as a colorless liquid. 1 H NMR:400MHz, CDCl3δ4.90-4.84(m,1H),3.40(t,J=6.8Hz,2H),2.30(t,J=7.2Hz,2H),1.90-1.82(m,2H) ,1.68-1.61(m,2H),1.52-1.45(m,5H),1.43-1.34(m,3H),1.30-1.25(m,20H),0.882(t,J=6.4Hz,6H)). [ka]

[0483] Preparation procedure for compound 5: Compound 4a (210 g, 2.36 mol, 219 mL, 30.0 eq) was added to a solution of compound 4 (32.0 g, 78.7 mmol, 1.00 eq) in 33 mL of EtOH at 25 °C. The mixture was heated to 65 °C and stirred at 65 °C for 16 h. The mixture was cooled to 25 °C and 100 mL of HO was added to the mixture. The reaction mixture was extracted with ethyl acetate (60.0 mL*3) and washed with saturated brine (50.0 mL*3) at 25 °C. The reaction mixture was dried over anhydrous NaSO at 25 °C. The organic phase was concentrated below 40-50 °C to obtain the residue. The residue was purified by column chromatography to obtain compound 5 (16.4 g, 65.5 mmol, 83.2% yield) as a red oil ( 1H NMR:400MHz, CDCl3δ4.89-4.83(m,1H),3.58-3.56(m,2H),2.66-2.62(m,2H),2.55-2.52(m,2H),2.25-2 .19(m,2H),1.64-1.53(m,6H),1.44-1.42(m,6H),1.28-1.25(m,7H),1.19(s,17H),0.89-0.86(m,6H)). [ka]

[0484] Preparation of compound 5a: Compound 5b (9.17 g, 47.0 mmol, 1.00 equiv.) was added to a solution of compound 5c (20.0 g, 51.7 mmol, 13.51 mL, 1.10 equiv.) in 400 mL of DCM at 25 °C. EDCI (11.7 g, 61.1 mmol, 1.30 equiv.), DMAP (1.15 g, 9.40 mmol, 0.200 equiv.), and DIEA (24.3 g, 188 mmol, 32.8 mL, 4.00 equiv.) were then added and the mixture was stirred at 25 °C for 16 h. 300 mL of HO was added to the mixture and the mixture was extracted with DCM (100 mL * 3) at 25 °C. The reaction mixture was dried over anhydrous NaSO and the organic phase was concentrated at 40-50 °C to obtain a residue. The residue was purified by column chromatography to give compound 5a (12.1 g, 21.5 mmol, 45.6% yield) as a white solid. 1 H NMR:EC3947-13-P1A1,400MHz,CDCl3δ5.38(d,J=4.8Hz,1H),4.67-4.59(m,1H),3.44-3.38(m,2H),2.34-2.29(m,4H),2.03-2.00(m,2H),1.66 -1.51(m,4H),1.50-1.48(m,6H),1.35-1.33(m,5H),1.27-1.13(m,7H), 1.03(s,3H),0.92(d,J=6.4Hz,3H),0.089-0.087(m,6H),0.07(s,3H)). [ka]

[0485] Procedure for preparing lipid target 1: Compound 6a (8.43 g, 14.9 mmol, 1.00 equiv.) was added to a solution of compound 6 (6.40 g, 14.9 mmol, 1.00 equiv.) in 140 mL of ACN at 25 °C. The solution was charged with K2CO3 (8.27 g, 59.9 mmol, 4.00 equiv.), KI (2.73 g, 16.5 mmol, 1.10 equiv.), and CPME (35.0 mL). The mixture was heated to 90 °C and stirred at 90 °C for 16 h. After cooling to 25 °C, the residue was dissolved in dichloromethane (100 mL) and filtered. The residue was purified by column chromatography to give compound 1 as a yellow oil (MS: m / z = 910.7 (M+H)). + , 1 H NMR:400MHz, CDCl3δ5.38(d,J=4.0Hz,1H),4.89-4.79(m,1H),4.64-4.58(m,1H),3 .55(d,J=8.0Hz,2H),2.45-2.35(m,6H),2.25-2.19(m,6H),1.98-1.88(m,2H),1.80 -1.72 (m, 3H), 1.57-1.52 (m, 8H), 1.47-1.34 (m, 13H), 1.27-1.14 (m, 31H), 1.10-1.01 (m, 8H), 0.95-0.90 (m, 6H), 0.89-0.82 (m, 5H), 0.81-0.78 (m, 10H), 0.68 (s, 3H). Figures 2A-2C show the chromatograms of compound target 1. 1 The H NMR spectrum (2A), mass spectrum (2B), and HPLC spectrum (2C) are shown.

[0486] Example 2: Synthesis scheme of ionizable lipid (compound target 11) [ka]

[0487] To a solution of compound 4a (10.0 g, 31.6 mmol, 1.00 equiv.) in DCM (270 mL) was added (COCl) (27.2 g, 214 mmol, 18.8 mL, 6.80 equiv.) and DMF (231 mg, 3.16 mmol, 243 μL, 0.01 equiv.). The mixture was stirred at 25° C. for 16 h. The reaction mixture was diluted with 3 drops of methanol to give a mixture, which was monitored by TLC. TLC (DCM:methanol=10:1) showed that compound 4a (R f =0.30) is completely consumed and a new spot (R f =0.80) was detected. The reaction mixture was then concentrated in vacuo to give compound 4b (9.00 g, crude) as a brown oil. [ka]

[0488] To a solution of compound 1 (4.00 g, 30.4 mmol, 1.00 equiv) in DCM (200 mL) was added (COCl) (11.6 g, 91.4 mmol, 8.01 mL, 3.00 equiv) and DMF (22.2 mg, 304 μmol, 23.4 μL, 0.01 equiv). The mixture was stirred at 25° C. for 16 h. The reaction mixture was diluted with 3 drops of methanol to give a mixture, which was monitored by TLC. TLC (DCM:methanol=10:1) showed that compound 1 (R f =0.00) is completely consumed and a new spot (R f =0.50) was detected. The reaction mixture was then concentrated in vacuo to give compound 1b (4.50 g, crude) as a brown solid. [ka]

[0489] To a solution of compound b1 (20.0 g, 153 mmol, 1.00 equiv.) in THF (400 mL) was added LiAlH4 (2.50 M, 61.4 mL, 1.00 equiv.) at 0 °C under N2. The mixture was stirred at 25 °C under N2 for 1 h. TLC (petroleum ether: EtOAc = 1:1) showed that compound b1 (R f=0.80) is consumed and a new spot (R f =0.20). The reaction mixture was quenched with Na2SO4·10H2O, which produced a large amount of white solid at 0 °C, and gas evolution ceased. The solid was filtered, and the filter cake was washed with DCM (3 × 100 mL). The combined filtrate was dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated in vacuo to give compound b2 (18.0 g, 136 mmol, 88.6% yield) as a colorless oil. 1 H NMR: (400MHz, DMSO-d6) δ4.94-4.91(m,1H), 3.80-3.74(m,2H), 3.52-3.44(m,3H), 1.33(s,3H), 1.27(s,3H). [ka]

[0490] To a solution of compound b2 (4.00 g, 30.2 mmol, 1.00 equiv.) and pyridine (4.79 g, 60.5 mmol, 4.89 mL, 2.00 equiv.) in DCM (200 mL) was added compound 4b (8.99 g, 30.2 mmol, 1.00 equiv.). The mixture was stirred at 25° C. for 2 h. TLC (petroleum ether: EtOAc = 1:1) showed that compound b2 (R f =0.20) is consumed and a new spot (R f =0.80) was detected. The reaction mixture was poured into 400 mL of water and then extracted with DCM (3 × 100 mL). The combined organic layers were washed with brine (100 mL), dried over NaSO, and filtered. The filtrate was concentrated in vacuo to give a residue. The residue was purified by column chromatography (SiO, petroleum ether: EtOAc = 100:1 to 5:1, Rf = 0.40 (petroleum ether: EtOAc = 5:1)) to give compound b3 (10.0 g, 23.1 mmol, 76.5% yield, 91.0% purity by LCMS on ELSD) as a pale yellow oil. 1H NMR:(400MHz,CDCl3)δ5.45-5.28(m,6H),4.74-4.69(m,1H),4.10(dd,J=13.2,3.6Hz,2H),3.81(dd,J=12.8,3.6Hz,2H),2.86-2 .75(m,4H),2.38(t,J=7.6Hz,2H),2.11-2.03(m,4H),1.68-1.60(m,2H),1.45(s,6H),1.36-1.30(m,8H),0.98(t,J=7.6Hz,3H). [ka]

[0491] A solution of compound b3 (10.0 g, 23.1 mmol, 1.00 equiv) in AcOH (80.0 mL) and HO (20.0 mL) was stirred at 50 °C for 1 h. TLC (petroleum ether: EtOAc = 1:1) confirmed that compound b3 (R f =0.80) is consumed and a new spot (R f =0.20) was detected. The reaction mixture was concentrated in vacuo and then diluted with ethyl acetate (100 mL). The organic layer was washed successively with saturated NaHCO3 solution (50.0 mL), water (50.0 mL), and brine (50.0 mL). The organic layer was then dried over Na2SO4 and filtered. The filtrate was concentrated in vacuo to give compound b4 (7.00 g, 18.9 mmol, 81.7% yield, 95.4% purity by LCMS with ELSD) as a yellow oil. 1 H NMR:(400MHz,DMSO-d6)δ5.40-5.24(m,6H),4.73-4.67(m,2H),4.02(dd,J=14.4,7.2Hz,1H),3.53-3.39(m,4H),2. 81-2.71(m,4H),2.30-2.24(m,2H),2.08-2.00(m,4H),1.55-1.47(m,2H),1.30-1.24(m,8H),0.92(t,J=7.2Hz,3H). [ka]

[0492] To a solution of compound b4 (7.00 g, 18.9 mmol, 1.00 equiv.) and pyridine (3.00 g, 37.8 mmol, 3.06 mL, 2.00 equiv.) in DCM (140 mL) was added a solution of compound 1b (2.27 g, 15.1 mmol, 0.80 equiv.) in DCM (20.0 mL). The mixture was stirred at 25 °C for 2 h. LCMS (EC15884-22-P1B1) showed that most of compound b4 (Rt = 2.118 min) was consumed and the desired mass (Rt = 1.845 min) was detected. The reaction mixture was poured into 200 mL of water and extracted with DCM (3 × 100 mL). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, and filtered. The filtrate was concentrated in vacuo to give a residue. The residue was purified by column chromatography (SiO, petroleum ether: EtOAc = 100:1 to 5:1, Rf = 0.40 (petroleum ether: EtOAc = 5:1)) to give compound 6 (3.50 g, 7.34 mmol, 38.7% yield, 97.7% purity by LCMS on ELSD) as a yellow oil. 1 H NMR:(400MHz,CDCl3)δ5.41-5.28(m,6H),5.11-5.04(m,1H),4.37-4.30(m,1H) ,4.25-4.18(m,1H),4.16-4.06(m,2H),2.84-2.74(m,4H),2.61-2.51(m,2H),2 .47-2.42(m,2H),2.40(d,J=4.4Hz,6H),2.36-2.30(m,2H),2.11-2.01(m,4H), 1.97-1.86(m,2H),1.65-1.57(m,2H),1.34-1.28(m,8H),0.96(t,J=7.6Hz,3H). [ka]

[0493] To a solution of compound 6 (3.20 g, 6.71 mmol, 1.00 equiv.) in DCM (60.0 mL) was added compound SM_1 (1.63 g, 8.06 mmol, 1.20 equiv.), EDCI (1.93 g, 10.0 mmol, 1.50 equiv.), DIEA (3.47 g, 26.8 mmol, 4.68 mL, 4.00 equiv.), and DMAP (164 mg, 1.34 mmol, 0.20 equiv.). The mixture was stirred at 25 °C for 12 h. LCMS (EC15884-24-P1A1) showed that compound 6 was completely consumed and the desired mass (Rt = 1.617 min) was detected. The reaction mixture was concentrated in vacuo to give a residue. The residue was diluted with 50.0 mL of ethyl acetate. The organic layer was washed with HO (50.0 mL), brine (50.0 mL), dried over NaSO, and filtered. The filtrate was concentrated in vacuo to give a residue. The residue was purified by column chromatography (SiO, DCM:methanol = 100:1 to 10:1, R f =0.50 (DCM:methanol=10:1)) to give compound 6d (3.00 g, 4.14 mmol, 61.6% yield, 89.6% purity by LCMS on ELSD) as a yellow oil. [ka]

[0494] To a solution of compound 6d (3.00 g, 4.14 mmol, 1.00 equiv.) in DCM (10.0 mL) was added HCl / dioxane (4.00 M, 30.0 mL, 29.0 equiv.). The mixture was stirred at 25 °C for 2 h. LCMS (EC15884-25-P1A1) showed that compound 6d was completely consumed and the desired mass (Rt = 0.334 min) was detected. The reaction mixture was concentrated in vacuo to give compound 6c (2.46 g, crude) as a yellow oil. [ka]

[0495] To a solution of compound 6c (2.46 g, 4.14 mmol, 1.00 equiv.) in DCM (25.0 mL) was added Int_7 (2.40 g, 6.21 mmol, 1.62 mL, 1.50 equiv.), EDCI (1.03 g, 5.39 mmol, 1.30 equiv.), DIEA (2.68 g, 20.7 mmol, 3.61 mL, 5.00 equiv.), and DMAP (101 mg, 828 μmol, 0.20 equiv.). The mixture was stirred at 25 °C for 12 h. LCMS (EC15884-26-P1A2) showed that compound 6c was completely consumed and the desired mass (Rt = 2.090 min) was detected. The reaction mixture was concentrated in vacuo to give a residue. The residue was diluted with 50.0 mL of ethyl acetate. The organic layer was washed with H2O (50.0 mL), brine (50.0 mL), dried over Na2SO4, and filtered. The filtrate was concentrated in vacuo to give target 11 (4.00 g, crude) as a yellow oil.

[0496] The crude target 11 (4.00 g, crude) was purified by preparative HPLC (column: Welch Ultimate XB-CN 250*50*10 μm, mobile phase: [EtOH+MeOH (4:1, neutral)], gradient: 12% to 100% B over 16 min), followed by column chromatography (SiO2, DCM:methanol = 100:0 to 50:1, R f = 0.35 (DCM:methanol = 10:1) to give target 11 (511.07 mg, 0.522 mmol, purity 98.3%, yield 12.5%) as a yellow oil. 1 H NMR:(400MHz,CDCl3)δ5.45-5.23(m,8H),4.67-4.57(m,1H),4.35-4.27(m,2H),4.19-4 .11(m,2H),2.86-2.75(m,4H),2.41-2.27(m,18H),2.13-1.94(m,7H),1.89-1.78(m,6H ),1.70-1.41(m,16H),1.39-1.33(m,5H),1.29-1.22(m,2H),1.21-1.04(m,8H),1.02(s ,3H),1.01-0.94(m,5H),0.92(d,J=6.4Hz,3H),0.87(d,J=6.8,1.6Hz,6H),0.68(s,3H).

[0497] Example 3. Synthesis scheme of ionizable lipid (compound target 12) [ka]

[0498] To a solution of compound Int_7 (11.0 g, 28.4 mmol, 7.43 mL, 1.10 equiv.) in DCM (220 mL) was added compound 1a (5.04 g, 25.8 mmol, 1.00 equiv.), EDCI (6.45 g, 33.6 mmol, 1.30 equiv.), DMAP (631 mg, 5.17 mmol, 0.20 equiv.), and DIEA (13.3 g, 103 mmol, 18.0 mL, 4.00 equiv.) at 25 °C. The mixture was stirred at 25 °C for 16 h. TLC (petroleum ether:ethyl acetate=10:1) showed that compound Int_7 (R f =0.10) is consumed, and a new spot (R f =0.50) was formed. The reaction mixture was concentrated in vacuo to give a residue. The residue was purified by column chromatography (SiO, petroleum ether:ethyl acetate = 100:1 to 10:1, Rf = 0.50 (petroleum ether:ethyl acetate = 10:1)) to give compound 1 (6.80 g, 12.0 mmol, 46.6% yield) as a white solid. 1 H NMR:(400MHz,CDCl3)δ5.38(d,J=4.0Hz,1H),4.67-4.56(m,1H),3.41(t,J=6.8 Hz,2H),2.36-2.27(m,4H),2.05-1.94(m,2H),1.93-1.81(m,5H),1.70-1.58(m, 4H),1.55-1.46(m,6H),1.45-1.27(m,5H),1.23-1.05(m,8H),1.02(s,3H),1.0 1-0.95(m,2H),0.92(d,J=6.4Hz,3H),0.87(dd,J=6.4,1.6Hz,6H),0.68(s,3H). [ka]

[0499] To a solution of compound 2a (1.14 g, 7.10 mmol, 1.00 equiv.) in ACN (60.0 mL) was added compound 1 (4.00 g, 7.10 mmol, 1.00 equiv.), K2CO3 (3.92 g, 28.3 mmol, 4.00 equiv.), KI (1.30 g, 7.81 mmol, 1.10 equiv.), and CPME (15.0 mL). The mixture was stirred at 90 °C for 12 h. LCMS (EC15884-12-P1B1) showed that compound 1 was completely consumed and the desired mass (Rt = 1.825 min) was detected. The reaction mixture was diluted with 100 mL of DCM and filtered. The filtrate was concentrated under vacuum to give the residue. The residue was purified by column chromatography (SiO2, DCM:methanol = 100:1 to 5:1, R f =0.35 (DCM:methanol=10:1)) to give compound 2 (3.00 g, 4.41 mmol, 62.1% yield, 94.7% purity by LCMS on ELSD) as a yellow oil. 1 H NMR:(400MHz,CDCl3)δ5.38(d,J=4.0Hz,1H),4.66-4.57(m,1H),3.61(t,J=5.6Hz,4H),2 .55-2.46(m,6H),2.35-2.26(m,4H),2.06-1.93(m,3H),1.90-1.79(m,4H),1.72-1.66(m, 3H),1.62-1.59(m,2H),1.58-1.40(m,10H),1.36-1.23(m,7H),1.22-1.04(m,9H),1.02(s ,3H),1.01-0.95(m,2H),0.92(d,J=6.4Hz,3H),0.87(dd,J=6.8,2.0Hz,6H),0.68(s,3H). [ka]

[0500] To a solution of compound 2 (2.00 g, 2.94 mmol, 1.00 equiv.) in DCM (20.0 mL) was added compound 3a (900 mg, 3.24 mmol, 1.10 equiv.), EDCI (732 mg, 3.82 mmol, 1.30 equiv.), DIEA (1.90 g, 14.7 mmol, 2.56 mL, 5.00 equiv.), and DMAP (71.8 mg, 588 μmol, 0.20 equiv.). The mixture was stirred at 25 °C for 12 h. LCMS (EC15884-14-P1A2) showed that most of compound 2 (Rt = 1.814 min) had been consumed, and the desired mass (Rt = 2.052 min) was detected. The reaction mixture was concentrated in vacuo to give a residue. The residue was purified by preparative HPLC (column: Welch Ultimate XB-NH2 250*50*10um, mobile phase: [EtOH+MeOH (4:1, neutral)], B%:1%, isocratic elution mode) to obtain target 12 (504.60mg, 0.530mmol, yield 18.0%, purity 95.0% by HPLC with ELSD) as an off-white oil. 1 H NMR:(400 MHz, CDCl3)δ5.44-5.28(m,7H),4.66-4.57(m,1H),4.07(t,J=6.4Hz,2H),3.60-3.54 (m,2H),2.85-2.75(m,4H),2.55-2.43(m,6H),2.34-2.26(m,7H),2.13-1.94(m,7H),1 .90-1.79(m,4H),1.69-1.43(m,28H),1.30-1.20(m,4H),1.19-1.05(m,8H),1.02(s,3 H),1.01-0.94(m,5H),0.92(d,J=6.4Hz,3H),0.87(d,J=6.4,1.6Hz,6H),0.68(s,3H).

[0501] Example 4. Synthesis scheme of ionizable lipid (compound target 13) [ka]

[0502] Compound 1 (30.0 g, 131 mmol, 1.00 equiv.) is placed in R1 (1.00 L three-bottom flask) at 20° C. DCM (350 mL) is placed in R1 at 20° C. Compound 1a (27.4 g, 131 mmol, 1.00 equiv.) is placed in R1 at 20° C. DIEA (67.9 g, 525 mmol, 91.5 mL, 4.00 equiv.), DMAP (3.21 g, 26.2 mmol, 0.20 equiv.), and EDCI (32.7 g, 170 mmol, 1.30 equiv.) are placed in R1 at 20° C. The mixture is stirred at 20° C. for 16 hours. TLC (petroleum ether:ethyl acetate=20:1) indicates that compound 1 (R f =0.60) is consumed, and the new main spot (R f =0.50) was detected. H2O (300 mL) was added to R2 (1.00 L barrel) at 20 °C. The reaction mixture was added to R2 at 20 °C. The reaction mixture was extracted with dichloromethane (300 mL * 3) at 20 °C. The reaction mixture was washed with saturated brine (200 mL * 2) at 20 °C. The reaction mixture was dried over anhydrous Na2SO4 at 20 °C. The organic phase was concentrated below 40-50 °C to obtain the residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 50:1 to 10:1, R f =0.50) to obtain compound 2 (30.0 g, 71.5 mmol, 54.4% yield) as a colorless liquid. [ka]

[0503] Compound 2 (30.0 g, 71.5 mmol, 1.00 equiv.) is placed in R1 (a 250 mL three-necked bottom flask) at 20° C. EtOH (30.0 mL) is placed in R1 at 20° C. Compound 2a (191 g, 2.15 mol, 199 mL, 30.0 equiv.) is placed in R1 at 20° C. The mixture is heated to 65° C. The mixture is stirred at 65° C. for 16 hours. TLC (dichloromethane:methanol=5:1) shows that compound 2 (R f =0.95) is consumed, and the new main spot (R f=0.20) was detected. H2O (100 mL) was added to R2 (250 mL barrel) at 20 °C. The reaction mixture was added to R2 at 20 °C. The reaction mixture was extracted with ethyl acetate (60.0 mL * 3) at 20 °C. The reaction mixture was washed with saturated brine (50.0 mL * 3) at 20 °C. The reaction mixture was dried over anhydrous Na2SO4 at 20 °C. The organic phase was concentrated below 40-50 °C to obtain the residue. The residue was purified by column chromatography (SiO2, dichloromethane:methanol = 50:1 to 10:1, R f =0.20) to obtain compound 3 (17.0 g, 39.7 mmol, 55.5% yield) as a pale yellow oil. [ka]

[0504] Compound 3-1 (5.00 g, 12.1 mmol, 1.10 equiv) is placed in R1 (2.00 L three-bottom flask) at 20 °C. DCM (100 mL) is placed in R1 at 20 °C. Compound 3-2 (2.15 g, 11.0 mmol, 1.00 equiv) is placed in R1 at 20 °C. EDCI (2.74 g, 14.3 mmol, 1.30 equiv), DIEA (5.69 g, 44.0 mmol, 7.67 mL, 4.00 equiv), and DMAP (269 mg, 2.20 mmol, 0.20 equiv) are placed in R1 at 20 °C. The mixture is stirred at 20 °C for 20 h. TLC (petroleum ether:ethyl acetate = 5:1) indicates that compound 3-1 (R f =0.30) is consumed, and the new main spot (R f =0.60) was detected. H2O (200 mL) was added to R2 (1.00 L barrel) at 20 °C. The reaction mixture was placed in R2 at 20 °C. The reaction mixture was extracted with dichloromethane (200 mL * 3) at 20 °C. The reaction mixture was dried over anhydrous Na2SO4 at 20 °C. The organic phase was concentrated below 40-50 °C to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 20:1 to 1:3, R f =0.60) to obtain compound 3-3 (3.30 g, 5.60 mmol, 50.8% yield) as a white solid. [ka]

[0505] Compound 3-3 (1.50 g, 2.54 mmol, 1.00 equiv.) is placed in R1 (500 mL three-neck barrel) at 20 °C. ACN (33.0 mL) is placed in R1 at 20 °C. Compound 3 (1.09 g, 2.54 mmol, 1.00 equiv.) is placed in R1 at 20 °C. K2CO3 (1.41 g, 10.1 mmol, 4 equiv.), KI (464 mg, 2.80 mmol, 1.10 equiv.), and CMPE (7.05 g, 70.4 mmol, 8.20 mL, 27.6 equiv.) are placed in R1 at 20 °C. The mixture is heated to 90 °C. The mixture is stirred at 90 °C for 16 h. TLC (dichloromethane:methanol = 5:1) shows that the reactants (R f =0.20) is consumed, and the new main spot (R f =0.50) was detected. H2O (100 mL) was added to R2 (1.00 L barrel) at 20 °C. The reaction mixture was placed in R2 at 20 °C. The reaction mixture was extracted with dichloromethane (100 mL * 3) at 20 °C. The reaction mixture was dried over anhydrous Na2SO4 at 20 °C. The organic phase was concentrated below 40-50 °C to obtain the residue. The residue was purified by column chromatography (SiO2, dichloromethane:methanol = 50:1 to 10:1, R f =0.50) to give target 13 (1.04 g, 1.11 mmol, 43.5% yield) as a colorless oil.

[0506] Example 5. Lipid nanoparticles for nucleic acid delivery Lipid nanoparticles (LNPs) play an important role in effectively protecting and delivering nucleic acids to cells for prophylactic and therapeutic applications. Despite encouraging data from ongoing clinical trials, the clinical use of gene therapy requires the discovery and development of more efficient delivery systems. Described herein are nanoparticles for gene and drug delivery.

[0507] EGFP mRNA, self-amplifying mRNA, SARS-CoV-2 mRNA lipid nanoparticles (LNPs).

[0508] Herein, for comparison, lipid nanoparticle (LNP) formulations were prepared using either Target 1 (ARV-T1), Target 11 (ARV-T11), Target 12 (ARV-T12), Target 13 (ARV-T13), or commercially available SM-102 lipids. LNP formulations were prepared using lipids dissolved in ethanol at a molar ratio of 50:10:38.5:1.5 (ionizable lipid:DSPC:cholesterol:PEG lipid). Using a NanoAssembler Ignite, the lipid mixture was combined with 100 mM sodium citrate buffer (pH 4.0) containing mRNA at a volumetric ratio of 3:1 (water:ethanol). The formulations were dialyzed against 10 mM Tris (pH 7.4) with 8% sucrose in a Slide-A-Lyzer dialysis cassette for at least 16 hours, concentrated using an Amicon ultracentrifugal filter, then passed through a 0.22 μm filter and stored at 4°C or -20°C until use. Size, PDI, and charge were measured by dynamic light scattering, and mRNA encapsulation efficiency was measured by Ribogreen assay spectroscopy using λex = 480 nm and λem = 535 nm.

[0509] Figure 3A shows the particle size and polydispersity index (PDI) of LNPs, and Figure 3B shows the surface charge (zeta potential) and mRNA encapsulation efficiency of SM-102 and ARV-T1 LNPs.

[0510] Plasmid DNA lipid nanoparticles (LNPs).

[0511] Herein, lipid nanoparticle (LNP) formulations were prepared using either target 1 (ARV-T1), target 11 (ARV-T11), target 12 (ARV-T12), or target 13 (ARV-T13) lipids. LNP formulations were prepared using lipids dissolved in ethanol containing supplemented DOTAP (10% molar ratio) lipids. Using a NanoAssembler Ignite, the lipid mixture was combined with 100 mM sodium citrate buffer (pH 4.0) containing mRNA at a volume ratio of 3:1 (water:ethanol). The formulations were dialyzed against 10 mM Tris (pH 7.4) with 8% sucrose in a Slide-A-Lyzer dialysis cassette for at least 16 hours, concentrated using an Amicon ultracentrifugal filter, then passed through a 0.22 μm filter and stored at 4 °C or -20 °C until use. The size, PDI, charge were measured by dynamic light scattering, and the mRNA encapsulation efficiency was measured by Ribogreen assay spectroscopy using λex=480 nm and λem=535 nm.

[0512] Figure 3A shows the particle size and polydispersity index (PDI) of LNPs, and Figure 3B shows the surface charge (zeta potential) and mRNA encapsulation efficiency of SM-102 and ARV-T1 LNPs.

[0513] In vitro studies 293T cells were transfected with mRNA encoding the SARS-CoV-2 spike protein. Figure 4 shows the in vitro expression of spike protein after transfection of cells with 0.5 and 1.0 μg / mL of mRNA over time.

[0514] Transfection efficiency was studied by delivering GFP mRNA (1 μg / mL) into BHK cells using LNP delivery. Transfection efficiency was determined 24 hours later by imaging the transfected BHK cells and analyzing GFP expression using flow cytometry. The results of the study are shown in Figures 5A and 5B.

[0515] In vivo studies LNPs formulated with the indicated ionizable lipids and 1 μg of luciferase-expressing mRNA were injected intramuscularly. Luciferase expression was determined by whole-body bioluminescence imaging using an IVIS Spectrum in vivo imaging system at 6, 24, 48, and 72 hours after administration. The results of the in vivo transfection efficiency study of luciferase-expressing mRNA are shown in Figure 6.

[0516] One microgram of vaccine formulated with the target 1 lipid (ARV-T1) and mRNA encoding the full-length spike glycoprotein of SARS-CoV-2 (δ mutant) was administered intramuscularly as scheduled. A formulation containing the commercially available lipid SM-102 and mRNA encoding the full-length spike glycoprotein of SARS-CoV-2 (δ mutant) was prepared and used for comparison. Total spike-specific IgG was assessed on days 14 and 35 after the first vaccination, and the results are shown in Figures 7B and 7C. Serum neutralizing antibodies were assessed using pseudotyped viruses, and the results are shown in Figure 7D. Antigen-specific T cell responses were assessed by Elispot, and the results are shown in Figure 7E. Data are presented as mean ± SD. Statistical comparisons were performed by one-way analysis of variance with Tukey's multiple comparison test. *p<0.05, **p<0.01. The results demonstrate efficient in vivo induction of immunity.

[0517] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the disclosed invention belongs. Publications cited herein and the materials for which they are cited are expressly incorporated by reference.

[0518] Those skilled in the art will recognize that numerous changes and modifications can be made to the preferred embodiments of the present invention and that such changes and modifications can be made without departing from the spirit of the present invention. It is, therefore, intended by the appended claims to cover all such equivalent variations as fall within the true spirit and scope of the present invention.

Claims

1. A compound of formula Y, 【Chemical 1】 or a pharmaceutically acceptable salt thereof, wherein: R １ is H or -CH ３ and 【Chemistry 2】 represents a carbon-carbon bond or a carbon-carbon double bond, R ７ is H or OR １６ and R ８ａ and R ８ｂ are each independently H or OR １６ or R ８ａ and R ８ｂ each taken together with the atom to which it is attached forms a cycloalkyl, aryl, heterocycloalkyl, or heteroaryl; R ９ H, C １ ~C ６ Alkyl, C ２ ~C ６ alkenyl, or C ２ ~C ６ is alkynyl, W is CR ４ａ or CR ４ａ R ４ｂ and when there is a double bond between W and the adjacent carbon, W is CR ４ａ and when there is a single bond between W and the adjacent carbon, W is CR ４ａ R ４ｂ and R ４ａ and R ４ｂ are each independently H, halogen, or C １ ~C ６ is alkyl, A is 【Chemistry 3】 and X is O or S; L １ is -O(C=O)-, -C(=O)-, -S(O) ｘ -, -C(=O)R ａ C(=O)-, -NR ａ C(=O)-, -NC(=O)R ａ C(=O)- or -C(=O)NR ａ C(═O)—, L ２ is -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -S(O) ｘ -, -SS-, -NR ａ C(=O)- or -C(=O)R ａ N-, L ３ does not exist, -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -NR ａ C(=O)- or -C(=O)R ａ N-, R ａ but 【Chemistry 4】 wherein n is an integer from 0 to 12; x is 0, 1 or 2; M is CH, 【Chemistry 5】 and G １ , G ２ and G ３ are each independently C １ ~C １２ Alkyl or C １ ~C １２ alkenyl, G ４ does not exist or C １ ~C １２ Alkyl or C １ ~C １２ alkenyl, R ２ Ga-CR ｂ R ｃ , C ６ ~C ２４ Alkyl or C ６ ~C ２４ is alkenyl, R ３ but 【Chemistry 6】 and R ６ がH、OR ｂ 、CN、-C(=O)OR ｂ 、-NC(=O)R ｂ 、-C(=O)NR ｂ 、 【Chemistry 7】 and R ｂ and R ｃ are independent and H, C １ ~C １２ Alkyl or C １ ~C １２ is alkenyl, p and q are independently 0, 1, 2, 3, or 4; R ３５ but 【Chemistry 8】 and L ４ does not exist or 【Chemistry 9】 and R １０ does not exist or C １ ~C ６ is alkyl, L ５ does not exist, or 【Chemistry 10】 and m is an integer of 1, 2, or 3; L ６ does not exist or 【Chemistry 11】 and R ４ is C ３ ~C １０ Alkyl, C ３ ~C １０ Cycloalkyl, C ３ ~C １０ Alkenyl, C ３ ~C １０ Cycloalkenyl, C ３ ~C １０ Alkynyl, C ３ ~C １０ Aryl, C ２ ~C ９ heterocyclyl, or C ２ ~C ９ is heteroaryl, L ７ is C １ ~C ６ is alkylene, R １３ａ , R １３ｂ and R １３ｃ are each independently C １ ~C ６ Alkyl or C ６ ~C １０ is aryl, R １４ , R １６ , R １９ , R ２１ , R ２２ , R ２４ , R ２８ , R ２９ and R ３１ are each independently H or C １ ~C ６ is alkyl, R １５ Ga-OR １６ , -NR １７ R １７ or 【Chemistry 12】 and R １７ are each independently H, -OR １６ , C ６ ~C １０ aryl, or C １ ~C ６ is alkyl, R １８ are each independently halogen or C １ ~C ６ is alkyl, o1 is an integer from 0 to 8, p1 and p2 each independently represent an integer of 0 to 2; Z is CH ２ , O, S or NR １６ and s and t are each independently 0 or 1; R ２３ Halo, hydroxyl, C １ ~C ６ Alkyl, or C １ ~C ６ is heteroalkyl, R ２０ , R ２５ａ and R ２５ｂ , R ３０ａ , R ３０ｂ , R ３０ｃ , R ３２ａ , R ３２ｂ and R ３４ are each independently C １ ~C ６ is alkyl, R ２６ａ and R ２６ｂ are independently H, C １ ~C ６ alkyl, or R ２６ａ and R ２６ｂ are combined with the atoms to which they are attached. 【Chemistry 13】 wherein R ２６ｃ and R ２６ｄ are each independently H or substituted C １ ~C ６ is alkyl, R ２７ａ and R ２６ｂ are each independently H, hydroxyl or C １ ~C ６ is alkyl, u is 1, 2 or 3; R ３３ａ and R ３３ｂ are each independently C １ ~C ６ Alkyl, C １ ~C ６ heteroalkyl, halogen, or hydroxyl; Q is O, S or NR １６ or a pharmaceutically acceptable salt thereof.

2. R ３５ but 【Chemistry 14】 where: L ４ does not exist or 【Chemistry 15】 and R １０ does not exist or C １ ~C ６ is alkyl, L ５ does not exist, or 【Chemistry 16】 and m is an integer of 1, 2, or 3; L ６ does not exist or 【Chemistry 17】 and R ４ is C ３ ~C １０ Alkyl, C ３ ~C １０ Cycloalkyl, C ３ ~C １０ Alkenyl, C ３ ~C １０ Cycloalkenyl, C ３ ~C １０ Alkynyl, C ３ ~C １０ Aryl, C ２ ~C ９ The compound of claim 1, which is heterocyclyl or C2-C9 heteroaryl.

3. A compound of formula I, 【Chemistry 18】 or a pharmaceutically acceptable salt thereof, wherein: R １ is H or -CH ３ and 【Chemistry 19】 represents a carbon-carbon bond or a carbon-carbon double bond, L ４ does not exist or 【Chemistry 20】 and R １０ does not exist or C １ ~C ６ is alkyl, L ５ does not exist, or 【Chemical 21】 and m is an integer of 1, 2, or 3; L ６ does not exist or 【Chemical 22】 and R ４ is C ３ ~C １０ Alkyl, C ３ ~C １０ Cycloalkyl, C ３ ~C １０ Alkenyl, C ３ ~C １０ Cycloalkenyl, C ３ ~C １０ Alkynyl, C ３ ~C １０ Aryl, C ２ ~C ９ Heterocyclyl, C ２ ~C ９ is heteroaryl, R ７ is H or OR １６ and R ８ａ and R ８ｂ are each independently H or OR １６ or R ８ａ and R ８ｂ each taken together with the atom to which it is attached forms a cycloalkyl, aryl, heterocycloalkyl, or heteroaryl; R １６ is H or C １ ~C ６ is alkyl, R ９ H, C １ ~C ６ Alkyl, C ２ ~C ６ alkenyl, or C ２ ~C ６ is alkynyl, W is CR ４ａ or CR ４ａ R ４ｂ and when there is a double bond between W and the adjacent carbon, W is CR ４ａ and when there is a single bond between W and the adjacent carbon, W is CR ４ａ R ４ｂ and R ４ａ and R ４ｂ are each independently H, halogen, or C １ ~C ６ is alkyl, A is 【Chemical 23】 and X is O or S; L １ is -O(C=O)-, -C(=O)-, -S(O) ｘ -, -C(=O)R ａ C(=O)-, -NR ａ C(=O)-, -NC(=O)R ａ C(=O)- or -C(=O)NR ａ C(═O)—, L ２ is -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -S(O) ｘ -, -SS-, -NR ａ C(=O)- or -C(=O)R ａ N-, L ３ does not exist, -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -NR ａ C(=O)- or -C(=O)R ａ N-, R ａ but 【Chemistry 24】 wherein n is an integer from 0 to 12; x is 0, 1 or 2; M is CH, 【Chemistry 25】 and G １ , G ２ and G ３ are each independently C １ ~C １２ Alkyl or C １ ~C １２ alkenyl, G ４ does not exist or C １ ~C １２ Alkyl or C １ ~C １２ alkenyl, R ２ Ga-CR ｂ R ｃ , C ６ ~C ２４ Alkyl or C ６ ~C ２４ is alkenyl, R ３ but 【Chemical 26】 and R ６ がH、OR ｂ 、CN、-C(=O)OR ｂ 、-NC(=O)R ｂ 、-C(=O)NR ｂ 、 【Chemical 27】 and R ｂ and R ｃ are independent and H, C １ ~C １２ Alkyl or C １ ~C １２ alkenyl, and A compound of Formula I or a pharmaceutically acceptable salt thereof, wherein p and q are independently 0, 1, 2, 3 or 4.

4. A compound of formula Ia, 【Chemical formula 28】 or a pharmaceutically acceptable salt thereof, R １ is H or -CH ３ and 【Chemical Formula 29】 represents a carbon-carbon bond or a carbon-carbon double bond, L ４ does not exist or 【Chemistry 30】 and L ５ does not exist, or 【Chemical 31】 and wherein m is an integer of 1, 2, or 3; L ６ does not exist or 【Chemical Formula 32】 and R ４ is C ３ ~C １０ Alkyl, C ３ ~C １０ Cycloalkyl, C ３ ~C １０ Alkenyl, C ３ ~C １０ Cycloalkenyl, C ３ ~C １０ Alkynyl, C ３ ~C １０ Aryl, C ２ ~C ９ Heterocyclyl, C ２ ~C ９ is heteroaryl, R １０ does not exist or C １ ~C ６ is alkyl, A is 【Chemical 33】 and X is O or S; L １ is -O(C=O)-, -C(=O)-, -S(O) ｘ -, -C(=O)R ａ C(=O)-, -NR ａ C(=O)-, -NC(=O)R ａ C(=O)- or -C(=O)NR ａ C(═O)—, L ２ is -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -S(O) ｘ -, -SS-, -NR ａ C(=O)- or -C(=O)R ａ N-, L ３ does not exist, -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -NR ａ C(=O)- or -C(=O)R ａ N-, R ａ but 【Chemical Formula 34】 wherein n is an integer from 0 to 12; x is 0, 1 or 2; M is CH, 【Chemical 35】 and G １ , G ２ and G ３ are each independently C １ ~C １２ Alkyl or C １ ~C １２ alkenyl, G ４ does not exist or C １ ~C １２ Alkyl or C １ ~C １２ alkenyl, R ２ Ga-CR ｂ R ｃ , C ６ ~C ２４ Alkyl or C ６ ~C ２４ is alkenyl, R ３ but 【Chemical 36】 and R ６ がH、OR ｂ 、CN、-C(=O)OR ｂ 、-NC(=O)R ｂ 、-C(=O)NR ｂ 、 【Chemical 37】 and R ｂ and R ｃ are independent and H, C １ ~C １２ Alkyl or C １ ~C １２ alkenyl, and A compound of Formula Ia, or a pharmaceutically acceptable salt thereof, wherein p and q are independently 0, 1, 2, 3 or 4.

5. The compound is 【Chemical Formula 38】 or a pharmaceutically acceptable salt thereof; In the formula, A, R １ , R ７ , R ８ａ , R ８ｂ , R ９ , W is the same as in formula Y.

6. The compound is 【Hua 39-1】 【Hua 39-2】 【Hua 39-3】 【Chemistry 39-4】 【Chemistry 39-5】 【Chemistry 39-6】 【Chemistry 39-7】 【Chemistry 39-8】 【Hua 39-9】 【Chemistry 39-10】 【Chemistry 39-11】 or a pharmaceutically acceptable salt thereof, or any combination thereof; The compound according to any one of claims 1 to 5, wherein A is the same as in formula Y.

7. The compound is 【Chemistry 40】 or a pharmaceutically acceptable salt thereof; The compound according to any one of claims 1 to 6, wherein A is the same as in formula Y.

8. a compound of formula II or formula III, 【Chemistry 41】 【Chemistry 42】 or a pharmaceutically acceptable salt thereof, wherein: R １ is H or -CH ３ and X is O or S; 【Chemistry 43】 represents a carbon-carbon bond or a carbon-carbon double bond, L ４ does not exist or 【Chemical 44】 and R １０ does not exist or C １ ~C ６ is alkyl, L ５ does not exist, or 【Chemistry 45】 and m is an integer of 1, 2, or 3; L ６ does not exist or 【Chemistry 46】 and R ４ is C ３ ~C １０ Alkyl, C ３ ~C １０ Cycloalkyl, C ３ ~C １０ Alkenyl, C ３ ~C １０ Cycloalkenyl, C ３ ~C １０ alkynyl, or derivatives thereof, L １ is -O(C=O)-, -C(=O)-, -S(O) ｘ -, -C(=O)R ａ C(=O)-, -NR ａ C(=O)-, -NC(=O)R ａ C(=O)- or -C(=O)NR ａ C(═O)—, L ２ is -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -S(O) ｘ -, -SS-, -NR ａ C(=O)- or -C(=O)R ａ N-, L ３ does not exist, -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -NR ａ C(=O)- or -C(=O)R ａ N-, R ａ but 【Chemistry 47】 wherein n is an integer from 0 to 12; x is 0, 1 or 2; M is CH, 【Chemistry 48】 and G １ , G ２ and G ３ are each independently C １ ~C １２ Alkyl or C １ ~C １２ alkenyl, G ４ does not exist or C １ ~C １２ Alkyl or C １ ~C １２ alkenyl, R ２ Ga-CR ｂ R ｃ , C ６ ~C ２４ Alkyl or C ６ ~C ２４ is alkenyl, R ３ but 【Chemistry 49】 and R ６ がH、OR ｂ 、CN、-C(=O)OR ｂ 、-NC(=O)R ｂ 、-C(=O)NR ｂ 、 【Chemistry 50】 and R ｂ and R ｃ are independent and H, C １ ~C １２ Alkyl or C １ ~C １２ alkenyl, and A compound of Formula II or Formula III, or a pharmaceutically acceptable salt thereof, wherein p and q are independently 0, 1, 2, 3, or 4.

9. R １ Ga-CH ３ The compound according to any one of claims 1 to 8, wherein

10. The compound of any one of claims 1 to 9, wherein X is O.

11. The compound according to any one of claims 1 to 10, wherein m is 3.

12. L ４ but 【Chemistry 51】 The compound according to any one of claims 1 to 11,

13. L ５ but 【Chemistry 52】 The compound according to any one of claims 1 to 12,

14. L ６ The compound according to any one of claims 1 to 13, wherein is absent.

15. R ４ The compound of any one of claims 1 to 14, wherein is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, pentyl, hexyl, heptyl, octyl, nonyl, or decyl.

16. R ４ The compound of any one of claims 1 to 15, wherein is isopropyl.

17. The compound of any one of claims 1 to 16, wherein M is CH.

18. R ３ but 【Chemistry 53】 The compound according to any one of claims 1 to 17,

19. R ６ がH、OR ｂ 、CN、-C(=O)OR ｂ 、-NC(=O)R ｂ 、-C(=O)NR ｂ 、 【Chemical 54】 The compound according to any one of claims 1 to 17,

20. a compound of formula IV or formula V, 【Chemistry 55】 【Chemical Formula 56】 or a pharmaceutically acceptable salt thereof, wherein: L １ is -O(C=O)-, -C(=O)-, -S(O) ｘ -, -C(=O)R ａ C(=O)-, -NR ａ C(=O)-, -NC(=O)R ａ C(=O)- or -C(=O)NR ａ C(═O)—, L ２ is -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -S(O) ｘ -, -SS-, -NR ａ C(=O)- or -C(=O)R ａ N-, L ３ does not exist, -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -NR ａ C(=O)- or -C(=O)R ａ N-, R ａ but 【Chemical 57】 wherein n is an integer from 0 to 12; x is 0, 1 or 2; M is CH, 【Chemistry 58】 and G １ , G ２ and G ３ are each independently C １ ~C １２ Alkyl or C １ ~C １２ alkenyl, G ４ does not exist or C １ ~C １２ Alkyl or C １ ~C １２ alkenyl, R ２ Ga-CR ｂ R ｃ , C ６ ~C ２４ Alkyl or C ６ ~C ２４ is alkenyl, R ３ but 【Chemical Formula 59】 and R ６ がH、OR ｂ 、CN、-C(=O)OR ｂ 、-NC(=O)R ｂ 、-C(=O)NR ｂ 、 【Chemistry 60】 and R ｂ and R ｃ are independent and H, C １ ~C １２ Alkyl or C １ ~C １２ alkenyl, and A compound of Formula IV or Formula V, or a pharmaceutically acceptable salt thereof, wherein p and q are independently 0, 1, 2, 3, or 4.

21. L １ The compound of claim 20, wherein is -C(=O)-.

22. L ２ The compound according to any one of claims 20 to 21, wherein is -(C=O)O-.

23. L ３ The compound according to any one of claims 20 to 22, wherein is absent.

24. G １ The compound according to any one of claims 20 to 23, wherein is a C5 alkyl.

25. G ２ The compound according to any one of claims 20 to 24, wherein is C6 alkyl.

26. G ３ The compound of any one of claims 20 to 25, wherein is C2 alkyl.

27. G ４ The compound of any one of claims 20 to 26, wherein is absent.

28. R ２ Ga-CR ｂ R ｃ The compound according to any one of claims 20 to 27,

29. R ３ but 【Hua 61】 The compound according to any one of claims 20 to 28, wherein

30. R ｂ and R ｃ is C ８ The compound of any one of claims 20 to 29, which is alkyl.

31. The compound according to any one of claims 20 to 30, wherein p and q are 0.

32. 32. The compound of any one of claims 20 to 31, wherein M is CH.

33. R ３ but 【Hua 62】 The compound according to any one of claims 20 to 32, wherein

34. R ６ がH、OR ｂ 、CN、-C(=O)OR ｂ 、-NC(=O)R ｂ 、-C(=O)NR ｂ 、 【Chemistry 63】 The compound according to any one of claims 20 to 32, wherein

35. The compound of formula Y, I, Ia or II-V is 【Hua 64-1】 【Hua 64-2】 【Hua 64-3】 【Hua 64-4】 【Hua 64-5】 【Hua 64-6】 【Hua 64-7】 or a pharmaceutically acceptable salt thereof.

36. The compound is 【Chemistry 65】 or a pharmaceutically acceptable salt thereof, or any combination thereof.

37. 1. A composition comprising: A compound according to any one of claims 1 to 36; and an active agent.

38. 38. The composition of claim 37, wherein the active agent is a diagnostic, prophylactic, or therapeutic agent.

39. The composition of any one of claims 37 to 38, wherein the active agent is a nucleic acid, a protein / peptide, a small molecule, or any combination thereof.

40. 40. The composition of claim 39, wherein the agent is a nucleic acid.

41. 41. The composition of any one of claims 37-40, wherein the nucleic acid comprises double-stranded DNA, single-stranded DNA, complexed DNA, encapsulated DNA, naked RNA, encapsulated RNA, messenger RNA (mRNA), tRNA, short interfering RNA (siRNA), double-stranded RNA (dsRNA), microRNA (miRNA), antisense RNA (asRNA), self-amplifying mRNA (saRNA), guide RNA (gRNA), cRNA, or any combination thereof.

42. The composition of any one of claims 37 to 41, wherein the nucleic acid comprises mRNA.

43. The mRNA may contain at least one of a metapneumovirus (hMPV) antigen polypeptide, a parainfluenza virus (hPIV) type 1, type 2, or type 3 antigen polypeptide (hPIV1, hPIV2, and hPIV3, respectively), a respiratory syncytial virus (RSV) antigen polypeptide, a measles virus (MeV) antigen polypeptide, a varicella-zoster antigen polypeptide, an influenza virus antigen polypeptide, a herpes simplex virus 1 (HSV1) antigen polypeptide, a herpes simplex virus 2 (HSV2) antigen polypeptide, a poxvirus (e.g., smallpox, monkeypox) antigen polypeptide, an African swine fever virus antigen polypeptide, a cytomegalovirus antigen polypeptide, an Epstein-Barr virus antigen polypeptide, a rotavirus antigen polypeptide, a rhino ...

43. The composition of claim 42, wherein the polypeptide encodes a human rabies virus antigen polypeptide, an adenovirus antigen polypeptide, a papillomavirus antigen polypeptide, a poliovirus antigen polypeptide, a mumps antigen polypeptide, a rabies antigen polypeptide, a rubella antigen polypeptide, a coxsackievirus antigen polypeptide, an equine encephalitis antigen polypeptide, a Japanese encephalitis antigen polypeptide, a yellow fever antigen polypeptide, a Rift Valley fever antigen polypeptide, a hepatitis A, B, C, D, or E virus antigen polypeptide, or a coronavirus (e.g., MERS-CoV, SARS-CoV, SARS-CoV2, HCoV-OC43, HCoV-229E, HCoV-NL63, HCoV-NL, HCoV-NH, HCoV-HKU1) antigen polypeptide, or any combination thereof.

44. 44. The composition of claim 43, wherein the coronavirus is a human coronavirus.

45. 45. The composition of claim 44, wherein the human coronavirus is selected from HCoV-229E, HCoV-OC43, HCoV-HKU1, HCoV-NL, HCoV-NH, HCoV-NL63, SARS-CoV, SARS-CoV-2, and MERS-CoV.

46. The composition of any one of claims 44 to 45, wherein the human coronavirus is SARS-CoV-2.

47. The composition of claim 42, wherein the mRNA encodes a tumor antigen, a cytokine (e.g., interferon, interleukin, colony-stimulating factor), a monoclonal antibody, or an antibody fragment (single-chain variable region fragment (scFv), fragment antigen binding ((Fab')2), intracellular antibody, nanobody), or any combination thereof.

48. 48. The composition of any one of claims 36 to 47, wherein the composition comprises a lipid nanoparticle dispersion, a liposomal formulation, a lipid emulsion, or any combination thereof.

49. A lipid nanoparticle, comprising: 20% to 80% of a compound according to any one of claims 1 to 36; 0% to 5% PEGylated lipid; 0% to 40% helper lipids; 0% to 80% sterols; and an active agent encapsulated in the nanoparticle.

50. 50. The nanoparticle of claim 49, wherein the active agent comprises a nucleic acid, a protein / peptide, a small molecule, or any combination thereof.

51. 51. The nanoparticle of claim 50, wherein the active agent is a nucleic acid.

52. 52. The nanoparticle of claim 51 , wherein the nucleic acid comprises double-stranded DNA, single-stranded DNA, complexed DNA, encapsulated DNA, naked RNA, encapsulated RNA, messenger RNA (mRNA), tRNA, short interfering RNA (siRNA), double-stranded RNA (dsRNA), microRNA (miRNA), antisense RNA (asRNA), self-amplifying mRNA (saRNA), guide RNA (gRNA), cRNA, or any combination thereof.

53. The nanoparticle of any one of claims 50 to 52, wherein the nucleic acid comprises mRNA.

54. 54. The nanoparticle of claim 53, wherein the mRNA encodes a tumor antigen, a cytokine (e.g., interferon, interleukin, colony-stimulating factor), a monoclonal antibody, or an antibody fragment (single-chain variable fragment (scFv), fragment antigen binding ((Fab')2), intracellular antibody, nanobody), or any combination thereof.

55. The mRNA may encode at least one of a human metapneumovirus (hMPV) antigen polypeptide, a parainfluenza virus (hPIV) type 1, type 2, and type 3 (hPIV1, hPIV2, and hPIV3, respectively) antigen polypeptide, a respiratory syncytial virus (RSV) antigen polypeptide, a measles virus (MeV) antigen polypeptide, a varicella-zoster antigen polypeptide, an influenza virus antigen polypeptide, a herpes simplex virus 1 (HSV1) antigen polypeptide, a herpes simplex virus 2 (HSV2) antigen polypeptide, a poxvirus (e.g., smallpox, monkeypox) antigen polypeptide, an African swine fever virus antigen polypeptide, a cytomegalovirus antigen polypeptide, an Epstein-Barr virus antigen polypeptide, a rotavirus antigen polypeptide, a rhino ...

54. The nanoparticle of claim 53, encoding a viral antigen polypeptide, an adenovirus antigen polypeptide, a papillomavirus antigen polypeptide, a poliovirus antigen polypeptide, a mumps antigen polypeptide, a rabies antigen polypeptide, a rubella antigen polypeptide, a coxsackievirus antigen polypeptide, an equine encephalitis antigen polypeptide, a Japanese encephalitis antigen polypeptide, a yellow fever antigen polypeptide, a Rift Valley fever antigen polypeptide, a hepatitis A, B, C, D, or E virus antigen polypeptide, or a coronavirus (e.g., MERS-CoV, SARS-CoV, SARS-CoV2, HCoV-OC43, HCoV-229E, HCoV-NL63, HCoV-NL, HCoV-NH, HCoV-HKU1) antigen polypeptide, or any combination thereof.

56. 56. The nanoparticle of claim 55, wherein the coronavirus is a human coronavirus.

57. 57. The nanoparticle of claim 56, wherein the human coronavirus is selected from HCoV-229E, HCoV-OC43, HCoV-HKU1, HCoV-NL, HCoV-NH, HCoV-NL63, SARS-CoV, SARS-CoV-2, and MERS-CoV.

58. The nanoparticle of any one of claims 55 to 57, wherein the human coronavirus is SARS-CoV-2.

59. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and an effective amount of lipid nanoparticles according to any one of claims 49 to 58.

60. 1. A method for delivering an active agent into a cell, comprising: A method comprising introducing into said cells a composition according to any one of claims 37 to 48 or a nanoparticle according to any one of claims 49 to 58.

61. A method for treating or preventing an infection caused by an infectious agent, comprising administering to a subject in need thereof a composition described in any one of claims 37 to 48, or a nanoparticle described in any one of claims 49 to 58.

62. A method for inducing an immune response against infection caused by an infectious agent, the method comprising administering to a subject in need thereof a composition according to any one of claims 37 to 48 or a nanoparticle according to any one of claims 49 to 58.

63. The infectious agent may be a bacterial organism, such as Streptococcus pneumoniae, Haemophilus influenzae, Staphylococcus aureus, Streptococcus pyrogenes, Corynebacterium diphtheriae, Listeria monocytogenes, Bacillus anthrax, Clostridium tetani, Clostridium botulinum, Clostridium perfringens, Neisseria meningitidis, Neisseria gonorrhoeae, Clostridium mutans, Pseudomonas aeruginosa, Salmonella typhi, Haemophilus influenzae, Bordetella pertussis, Francisella tularensis, Yersinia pestis, Vibrio cholerae, Legionella pneumophila, Mycobacterium tuberculosis, Mycobacterium leprae, Treponema pallidum, Bordetella interrogans, Borrelia burgdorferi, Campylobacter jejuni, or the like; a virus, such as Metapneumonia, viruses, such as human metapneumovirus (hMPV), parainfluenza viruses, such as human parainfluenza virus (hPIV) types 1, 2, and 3 (hPIV1, hPIV2, and hPIV3, respectively), respiratory syncytial virus (RSV), measles virus (MeV), coronaviruses (e.g., MERS-CoV, SARS-CoV, SARS-CoV2, HCoV-OC43, HCoV-229E, HCoV-NL63, HCoV-NL, HCoV-NH, and HCoV-HKU1), poxviruses, viruses (e.g., smallpox, monkeypox), African swine virus, influenza A and B, human immunodeficiency virus (HIV), varicella-zoster, herpes simplex types 1 and 2, cytomegalovirus, Epstein-Barr virus, rotavirus, rhinovirus, adenovirus, papillomavirus, poliovirus, mumps, rabies, rubella, coxsackievirus, equine encephalitis, Japanese encephalitis, yellow fever, Rift Valley fever, hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis D virus, hepatitis E virus, and the like; 63. The method of any one of claims 61 to 62, comprising fungi, protozoa and parasites such as Cryptococcus neoformans, Histoplasma capsulatum, Candida albicans, Candida tropicalis, Nocardia asteroides, Rickettsia rickettsii, Salmonella typhi, Mycoplasma pneumoniae, Chlamydia psittaci, Chlamydia trachomatis, Plasmodium falciparum, Trypanosoma brucei, Entamoeba histolytica, Toxoplasma gondii, Trichomonas protozoa, Schistosoma mansoni and the like.

64. A method for treating or preventing a respiratory infection, comprising administering to a subject in need thereof a composition according to any one of claims 37 to 48, or a nanoparticle according to any one of claims 49 to 58.

65. A method for inducing an immune response against a respiratory virus, comprising administering to a subject in need thereof a composition according to any one of claims 37 to 48, or a nanoparticle according to any one of claims 49 to 58.

66. 66. The method of claim 65, wherein the respiratory infection is caused by infection with metapneumovirus (MPV), human parainfluenza virus (hPIV) types 1, 2, and 3 (hPIV1, hPIV2, and hPIV3, respectively), RSV, measles virus (MeV), or coronavirus.

67. 67. The method of claim 66, wherein the coronavirus is a human coronavirus.

68. 68. The method of claim 67, wherein the human coronavirus is selected from HCoV-229E, HCoV-OC43, HCoV-HKU1, HCoV-NL63, SARS-CoV, SARS-CoV-2, and MERS-CoV.

69. 69. The method of any one of claims 67 to 68, wherein the human coronavirus is SARS-CoV-2.

70. 70. The method of any one of claims 61 to 69, wherein the subject is a veterinary patient.

71. 71. The method of any one of claims 61 to 70, wherein the subject is a mammal.

72. 72. The method of claim 71, wherein the mammal is a human.

73. 73. The method of any one of claims 60-72, which may be administered by oral, topical, transdermal, transcutaneous, intra-articular, intra-arteriolar, intradermal, intraventricular, intralesional, intranasal, rectal, vaginal, inhalation, implanted reservoir, subcutaneous, intravenous, intramuscular, intra-articular, intra-synovial, intrasternal, intrathecal, intraperitoneal, intrahepatic, intralesional, intracranial injection or infusion.

74. 1. A method for delivering an active agent into a cell, comprising: A method comprising introducing into said cells a composition according to any one of claims 37 to 48 or a nanoparticle according to any one of claims 49 to 58.

75. 75. The method of claim 74, wherein the composition or nanoparticle comprises an active agent comprising a nucleic acid, a protein / peptide, a small molecule, or any combination thereof.

76. 76. The method of claim 75, wherein the active agent comprises a nucleic acid.

77. 77. The method of any one of claims 74 to 76, comprising introducing a composition or nanoparticle comprising a nucleic acid into a specific cell or tissue, expressing a protein encoded by said nucleic acid, and correcting a defect caused by a deficiency of said nucleic acid in said cell or tissue.

78. 77. The method of any one of claims 74 to 76, comprising introducing a composition or nanoparticle comprising a nucleic acid into specific cells or tissues in vivo, expressing a protein encoded by the nucleic acid in the specific cells or tissues, and inducing an immune response by the subject.

79. introducing a composition or nanoparticles containing the nucleic acid into specific cells or tissues ex vivo, and expressing the protein encoded by the nucleic acid in the specific cells or tissues to generate chimeric cells or tissues; and administering said melanoma cells or tissue to a subject.

80. The method of any one of claims 74 to 76, wherein the specific cells are T cells (e.g., CAR T cells) derived from the subject.

81. 77. The method of any one of claims 74 to 76, wherein the method introduces a polynucleotide to modulate gene expression, protein expression, or any combination thereof.

82. 77. The method of any one of claims 74 to 76, wherein the method introduces a polynucleotide for cell reprogramming by modulating cell behavior.

83. 77. The method of any one of claims 74 to 76, wherein the polynucleotide encodes for an intracellular antibody.

84. 77. The method of any one of claims 74 to 76, comprising introducing a composition or nanoparticle comprising a nucleic acid into a specific cell or tissue, silencing expression of a protein encoded by said nucleic acid, and correcting a defect caused by overexpression of said nucleic acid in said cell or tissue.

85. 77. The method of any one of claims 74 to 76, comprising introducing a composition or nanoparticle comprising a nucleic acid into a specific cell or tissue, silencing expression of a protein encoded by the nucleic acid in said specific cell or tissue, and inducing an immune response by said subject.

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